LOCUS       Exported                4361 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBR322, complete sequence.
ACCESSION   J01749
VERSION     .
KEYWORDS    ampicillin resistance; beta-lactamase; cloning vector; drug
            re...
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4361)
  AUTHORS   Sutcliffe JG.
  TITLE     Nucleotide sequence of the ampicillin resistance gene of Escherichia
            coli plasmid pBR322
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 75 (8), 3737-3741 (1978)
  PUBMED    358200
REFERENCE   2  (bases 1 to 4361)
  AUTHORS   Sutcliffe JG.
  TITLE     Complete nucleotide sequence of the Escherichia coli plasmid pBR322
  JOURNAL   Cold Spring Harb. Symp. Quant. Biol. 43 (Pt 1), 77-90 (1979)
  PUBMED    383387
REFERENCE   3  (bases 1 to 4361)
  AUTHORS   Reed RR, Young RA, Steitz JA, Grindley ND, Guyer MS.
  TITLE     Transposition of the Escherichia coli insertion element gamma 
            generates a five-base-pair repeat
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 76 (10), 4882-4886 (1979)
  PUBMED    388421
REFERENCE   4  (bases 1 to 4361)
  AUTHORS   Covarrubias L, Cervantes L, Covarrubias A, Soberon X, Vichido I, 
            Blanco A, Kupersztoch-Portnoy YM, Bolivar F.
  TITLE     Construction and characterization of new cloning vehicles. V. 
            Mobilization and coding properties of pBR322 and several deletion 
            derivatives including pBR327 and pBR328
  JOURNAL   Gene 13 (1), 25-35 (1981)
  PUBMED    6263753
REFERENCE   5  (bases 1 to 4361)
  AUTHORS   Marians KJ, Soeller W, Zipursky SL.
  TITLE     Maximal limits of the Escherichia coli replication factor Y effector
            site sequences in pBR322 DNA
  JOURNAL   J. Biol. Chem. 257 (10), 5656-5662 (1982)
  PUBMED    6279609
REFERENCE   6  (bases 1 to 4361)
  AUTHORS   Brosius J, Cate RL, Perlmutter AP.
  TITLE     Precise location of two promoters for the beta-lactamase gene of 
            pBR322. S1 mapping of ribonucleic acid isolated from Escherichia 
            coli or synthesized in vitro
  JOURNAL   J. Biol. Chem. 257 (15), 9205-9210 (1982)
  PUBMED    6178738
REFERENCE   7  (bases 1 to 4361)
  AUTHORS   Van Dyke MW, Hertzberg RP, Dervan PB.
  TITLE     Map of distamycin, netropsin, and actinomycin binding sites on 
            heterogeneous DNA: DNA cleavage-inhibition patterns with 
            methidiumpropyl-EDTA.Fe(II)
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 79 (18), 5470-5474 (1982)
  PUBMED    6291045
REFERENCE   8  (bases 1 to 4361)
  AUTHORS   Peden KW, Nathans D.
  TITLE     Local mutagenesis within deletion loops of DNA heteroduplexes
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 79 (23), 7214-7217 (1982)
  PUBMED    6760191
REFERENCE   9  (bases 1 to 4361)
  AUTHORS   Peden KW.
  TITLE     Revised sequence of the tetracycline-resistance gene of pBR322
  JOURNAL   Gene 22 (2-3), 277-280 (1983)
  PUBMED    6307828
REFERENCE   10  (bases 1 to 4361)
  AUTHORS   Watabe H, Iino T, Kaneko T, Shibata T, Ando T.
  TITLE     A new class of site-specific endodeoxyribonucleases. Endo.Sce I 
            isolated from a eukaryote, Saccharomyces cerevisiae
  JOURNAL   J. Biol. Chem. 258 (8), 4663-4665 (1983)
  PUBMED    6300094
REFERENCE   11  (bases 1 to 4361)
  AUTHORS   Livneh Z.
  TITLE     Directed mutagenesis method for analysis of mutagen specificity: 
            application to ultraviolet-induced mutagenesis
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 80 (1), 237-241 (1983)
  PUBMED    6337373
REFERENCE   12  (bases 1 to 4361)
  AUTHORS   Mascharak PK, Sugiura Y, Kuwahara J, Suzuki T, Lippard SJ.
  TITLE     Alteration and activation of sequence-specific cleavage of DNA by 
            bleomycin in the presence of the antitumor drug 
            cis-diamminedichloroplatinum(II)
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 80 (22), 6795-6798 (1983)
  PUBMED    6196777
REFERENCE   13  (bases 1 to 4361)
  AUTHORS   Schultz PG, Dervan PB.
  TITLE     Sequence-specific double-strand cleavage of DNA by 
            penta-N-methylpyrrolecarboxamide-EDTA X Fe(II)
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 80 (22), 6834-6837 (1983)
  PUBMED    6417654
REFERENCE   14  (bases 1 to 4361)
  AUTHORS   Sutcliffe JG.
  JOURNAL   Unpublished
REFERENCE   15  (bases 1 to 4361)
  AUTHORS   Fuller RS, Funnell BE, Kornberg A.
  TITLE     The dnaA protein complex with the E. coli chromosomal replication 
            origin (oriC) and other DNA sites
  JOURNAL   Cell 38 (3), 889-900 (1984)
  PUBMED    6091903
REFERENCE   16  (bases 1 to 4361)
  AUTHORS   Lathe R, Kieny MP, Skory S, Lecocq JP.
  TITLE     Linker tailing: unphosphorylated linker oligonucleotides for joining
            DNA termini
  JOURNAL   DNA 3 (2), 173-182 (1984)
  PUBMED    6327214
REFERENCE   17  (bases 1 to 4361)
  AUTHORS   Heusterspreute M, Davison J.
  TITLE     Restriction site bank vectors. II. DNA sequence analysis of plasmid 
            pJRD158
  JOURNAL   DNA 3 (3), 259-268 (1984)
  PUBMED    6086259
REFERENCE   18  (bases 1 to 4361)
  AUTHORS   Abarzua P, Soeller W, Marians KJ.
  TITLE     Mutational analysis of primosome assembly sites. I. Distinct classes
            of mutants in the pBR322 Escherichia coli factor Y DNA effector 
            sequences
  JOURNAL   J. Biol. Chem. 259 (22), 14286-14292 (1984)
  PUBMED    6209275
REFERENCE   19  (bases 1 to 4361)
  AUTHORS   Soeller W, Abarzua P, Marians KJ.
  TITLE     Mutational analysis of primosome assembly sites. II. Role of 
            secondary structure in the formation of active sites
  JOURNAL   J. Biol. Chem. 259 (22), 14293-14300 (1984)
  PUBMED    6150042
REFERENCE   20  (bases 1 to 4361)
  AUTHORS   Van Dyke MM, Dervan PB.
  TITLE     Echinomycin binding sites on DNA
  JOURNAL   Science 225 (4667), 1122-1127 (1984)
  PUBMED    6089341
REFERENCE   21  (bases 1 to 4361)
  AUTHORS   Pouwels PH, Enger-Valk BE, Brammar WJ.
  JOURNAL   (in) CLONING VECTORS. Elsevier Scientific Publishing, Amsterdam 
            (1985)
REFERENCE   22  (bases 1 to 4361)
  AUTHORS   Watson N.
  TITLE     A new revision of the sequence of plasmid pBR322
  JOURNAL   Gene 70 (2), 399-403 (1988)
  PUBMED    3063608
REFERENCE   23  (bases 1 to 4361)
  AUTHORS   Gilbert W.
  TITLE     Obtained from VecBase 3.0
  JOURNAL   Unpublished
REFERENCE   24  (bases 1 to 4361)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On or before Apr 4, 2002 this sequence version replaced L08654.1, 
            gi:58257.
            The circular sequence is numbered such that 0 is the middle of the 
            unique EcoRI site and the count increases first through the tet 
            genes, the pMB1 material, and finally through the Tn3 region. 
            Plasmid pBR322 contains ampicillin and tetracycline resistance 
            genes.  The ampicillin resistance gene (amp-r) is a penicillin 
            beta-lactamase. Promoters P1 and P3 are for the beta-lactamase gene.
            P3 is the natural promoter, and P1 is artificially created by the 
            ligation of two different DNA fragments to create pBR322. P2 is in 
            the same region as P1, but it is on the opposite strand and 
            initiates transcription in the direction of the tetracycline 
            resistance gene.
            Mutational studies in the primosome assembly sites indicate four 
            types of mutations:  Class I having no effect on the activities 
            elicited by the DNA site and the bases involved are probably 
            spacers; Class II requiring higher Mg-2+ concentrations than the 
            wild-type to be fully activated as factor Y ATPase effectors; Class 
            III co-inactivating both the ATPase effector and DNA replication 
            template activity of the site, indicating that they probably 
            represent essential contact points between factor Y and the DNA; 
            Class IV having a replication template activity intermediate that of
            class III and class II mutant DNAs.
            Specific sites within or near the origins of replication are 
            recognized by dnaA protein.  Without dnaA binding to the origin of 
            replication chromosomal replication is not possible (15). pBR322 DNA
            contains two separate regions on opposite strands and close to the 
            origin of replication which, when in single-stranded form, can act 
            as effectors for the ATPase activity of E.coli replication factor Y 
            (5).  Small fragments of DNA containing these sites when cloned in 
            an f1 phage vector act as origins of DNA replication allowing the 
            formation of complementary double-stranded DNA in 
            rifampicin-resistant, dna(B,G,C)-dependent fashion in vitro (5). The
            biological activity of echinomycin is thought to be related to the 
            formation of complexes by intercalating with cellular DNA (20). 
            Complete source information:
            Plasmid pBR322 from E.coli (2),(1),(3),(6),(11),(8),(5),(7),(12), 
            (13),(10),(9),(14),(18),(19),(15),(20),(16); pBR322 DNA in pXf3 (4).
            The following data and their annotation were supplied by Will 
            Gilbert under the auspices of the Curator Program.
             CROSSREFERENCE
                           #parent
                             GenBank(50):pSC101C, GenBank(50):Trn3
                           #offspring
                             VecBase(3):pBR325, VecBase(3):pBR327, 
            VecBase(3):pBR328,
                             VecBase(3):pAT153, VecBase(3):pUC7, 
            VecBase(3):pJRD158,
                             VecBase(3):PiVX, VecBase(3):PiAN7, 
            VecBase(3):pSP64,
                             VecBase(3):pSP65, VecBase(3):pGEM1, 
            VecBase(3):pGEM2,
                             VecBase(3):pGEM3, VecBase(3):pGEM4, 
            VecBase(3):pKK223,
                             VecBase(3):pLBU3, VecBase(3):pTrS3, 
            VecBase(3):pRSVNeo,
                             VecBase(3):pSV2Cat, VecBase(3):M13mp9, 
            VecBase(3):pHC79,
                             VecBase(3):pV34, VecBase(3):pKTH601, 
            VecBase(3):pKTH604,
                             VecBase(3):pKTH605, VecBase(3):pKTH606, 
            VecBase(3):YEp24,
                             VecBase(3):YIp5, VecBase(3):YRp17, 
            VecBase(3):pSP18,
                        VecBase(3):pSP19,
                             VecBase(3):pSP6T3, VecBase(3):pSP6T719, 
            VecBase(3):pT712,
                             VecBase(3):pT713, VecBase(3):pT7T318, 
            VecBase(3):pT7T319,
                             VecBase(3):pT7T3A18, VecBase(3):pT7T3A19, 
            VecBase(3):pEX1,
                             VecBase(3):pEX2, VecBase(3):pEX3, 
            VecBase(3):pCKSP6,
                             VecBase(3):pACYC177, VecBase(3):pKO1, 
            VecBase(3):pKO2,
                        VecBase(3):pKM1,
                             VecBase(3):pKM2, VecBase(3):pMBL1, 
            VecBase(3):pMBL604,
                             VecBase(3):pMC1511, VecBase(3):pMC1871, 
            VecBase(3):pAA37X,
                             VecBase(3):pUR278, VecBase(3):pUR288, 
            VecBase(3):pUR289,
                             VecBase(3):pUR290, VecBase(3):pUR291, 
            VecBase(3):pUR292,
                             VecBase(3):pUR222.
FEATURES             Location/Qualifiers
     source          1..4361
                     /organism="Cloning vector pBR322"
                     /focus
                     /mol_type="other DNA"
                     /tissue_lib="ATCC 31344, ATCC 37017"
                     /db_xref="taxon:47470"
     source          1..1762
                     /organism="Plasmid pSC101"
                     /mol_type="other DNA"
                     /db_xref="taxon:2625"
     promoter        10..38
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     misc_binding    24..27
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     regulatory      complement(27..33)
                     /regulatory_class="promoter"
                     /note="promoter P1 (6)"
     misc_binding    39..42
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     regulatory      43..49
                     /regulatory_class="promoter"
                     /note="promoter P2 (6)"
     misc_binding    53..56
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     misc_binding    67..70
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     misc_binding    80..83
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     CDS             86..1276
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline resistance protein"
                     /label=tet
                     /protein_id="AAB59735.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            86..1276
                     /gene="tet"
                     /label=tet
     misc_feature    complement(141..142)
                     /note="Endo.Sce I cleavage site coordinated with site at 
                     base 146 (10)"
     misc_feature    146..147
                     /gene="tet"
                     /note="Endo.Sce I cleavage site coordinated with site at 
                     base 142 (10)"
     misc_binding    411..414
                     /gene="tet"
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     misc_difference 426
                     /gene="tet"
                     /label=conflict
                     /note="conflict"
                     /citation="[11]"
     misc_binding    469..472
                     /gene="tet"
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     old_sequence    526..528
                     /gene="tet"
                     /citation="[17]"
     repeat_region   1515..1519
                     /label=gamma-delta insertion target sequence
                     /note="gamma-delta insertion target sequence"
     misc_feature    1636..1762
                     /label=from pSC101 (bp 1860-1986)
                     /note="from pSC101 (bp 1860-1986)"
     repeat_region   1788..1792
                     /label=gamma-delta insertion target sequence
                     /note="gamma-delta insertion target sequence"
     misc_difference 1891..1892
                     /replace="att"
                     /label=conflict
                     /note="conflict"
                     /citation="[23]"
     old_sequence    1892..1893
                     /citation="[2]"
                     /citation="[22]"
     regulatory      1905..1910
                     /regulatory_class="ribosome_binding_site"
     regulatory      1905..1909
                     /regulatory_class="ribosome_binding_site"
                     /note="Shine-Dalgarno sequence"
     misc_difference 1913..1914
                     /replace="caa"
                     /label=conflict
                     /note="conflict"
                     /citation="[23]"
     old_sequence    1914..1915
                     /citation="[17]"
     CDS             1915..2106
                     /codon_start=1
                     /product="ROP protein"
                     /label=ROP protein
                     /protein_id="AAB59736.1"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2011..2167
                     /label=H-strand Y effector site
                     /note="H-strand Y effector site"
                     /citation="[5]"
     misc_feature    2208..2348
                     /label=bom
                     /note="basis of mobility region from pBR322"
     repeat_region   2245..2249
                     /label=gamma-delta insertion target sequence
                     /note="gamma-delta insertion target sequence"
     misc_feature    complement(2351..2414)
                     /label=L-strand Y effector site
                     /note="L-strand Y effector site"
                     /citation="[5]"
     misc_binding    2439..2447
                     /label=dnaA binding site
                     /bound_moiety="dnaA"
     rep_origin      complement(2534..3122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2535
     old_sequence    2729..2730
                     /replace="at"
                     /label=revision according to (17)
                     /note="revision according to (17)"
                     /citation="[2]"
                     /citation="[17]"
     old_sequence    2729
                     /citation="[17]"
     old_sequence    2730
                     /replace="t"
                     /label=revision according to (16)
                     /note="revision according to (16)"
                     /citation="[2]"
                     /citation="[16]"
                     /citation="[17]"
     mobile_element  3148..4361
                     /mobile_element_type="transposon:Tn3"
     repeat_region   3148..3185
                     /note="corresponds to one of the 38bp repeats found in Tn3 
                     (bp 1-38 and complement (4920-4957))"
     CDS             complement(3293..4153)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="E-286"
                     /protein_id="AAB59737.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3293..4153)
                     /gene="bla"
                     /label=bla
     mat_peptide     complement(3296..4084)
                     /gene="bla"
                     /product="beta-lactamase"
     sig_peptide     complement(4085..4153)
                     /gene="bla"
     promoter        complement(4154..4258)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(4161..4165)
                     /regulatory_class="ribosome_binding_site"
                     /note="Shine-Dalgarno sequence"
     regulatory      complement(4188..4194)
                     /regulatory_class="promoter"
                     /note="promoter P3 (6)"
     misc_binding    complement(4268..4271)
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     misc_binding    complement(4280..4283)
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     misc_binding    complement(4285..4288)
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     misc_binding    complement(4296..4299)
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     misc_binding    complement(4311..4314)
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     misc_binding    complement(4317..4320)
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"
     misc_binding    complement(4331..4334)
                     /label=echinomycin binding site
                     /bound_moiety="echinomycin"