LOCUS       Exported                5321 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWD42-09, complete sequence.
ACCESSION   AF129072
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5321)
  AUTHORS   Larsen JE, Gerdes K, Light J, Molin S.
  TITLE     Low-copy-number plasmid-cloning vectors amplifiable by derepression 
            of an inserted foreign promoter
  JOURNAL   Gene 28 (1), 45-54 (1984)
  PUBMED    6329915
REFERENCE   2  (bases 1 to 5321)
  AUTHORS   Weiss RB, Dunn DM.
  TITLE     Copy-number inducible, medium-insert plasmid-cloning vectors for 
            sub-clone library construction and DNA sequencing
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5321)
  AUTHORS   Weiss RB, Dunn DM.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-FEB-1999) Human Genetics, University of Utah, 20 South
            2030 East, Salt Lake City, Utah 84112, USA
REFERENCE   4  (bases 1 to 5321)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5321
                     /organism="Cloning vector pWD42-09"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /label=copy-number inducible plasmid R1 vector
                     /note="copy-number inducible plasmid R1 vector"
                     /db_xref="taxon:92833"
     CDS             complement(74..787)
                     /codon_start=1
                     /gene="lambda cI857 repressor"
                     /product="repressor protein"
                     /label=lambda cI857 repressor
                     /protein_id="AAD28615.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     gene            complement(74..787)
                     /gene="lambda cI857 repressor"
                     /label=lambda cI857 repressor
     regulatory      835..870
                     /regulatory_class="promoter"
                     /note="over-expression of RepA at 42oC; runaway plasmid 
                     replication; lambda pR"
     RBS             876..884
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             1040..1300
                     /codon_start=1
                     /gene="CopB"
                     /product="CopB protein"
                     /label=CopB
                     /protein_id="AAD28614.1"
                     /translation="MSQTENAVTSSLSQKRFVRRGKPMTDSEKQMAAVARKRLTHKEIK
                     VFVKNPLKDLMVEYCEREGITQAQFVEKIIKDELQRLDILK"
     gene            1040..1300
                     /gene="CopB"
                     /label=CopB
     misc_feature    complement(1416..1507)
                     /label=antisense RNA for CopA
                     /note="antisense RNA for CopA"
     CDS             1592..2449
                     /codon_start=1
                     /gene="RepA"
                     /product="replication protein A"
                     /label=RepA
                     /protein_id="AAD28613.1"
                     /translation="MTDLHQTYYRQVKNPNPVFTPREGAGTLKFCEKLMEKAVGFTSRF
                     DFAIHVAHARSRGLRRRMPPVLRRRAIDALLQGLCFHYDPLANRVQCSITTLAIECGLA
                     TESGAGKLSITRATRALTFLSELGLITYQTEYDPLIGCYIPTDITFTLALFAALDVSED
                     AVAAARRSRVEWENKQRKKQGLDTLGMDELIAKAWRFVRERFRSYQTELKSRGIKRARA
                     RRDANRERQDIVTLVKRQLTREISEGRFTANGEAVKREVERRVKERMILSRNRNYSRLA
                     TASP"
     gene            1592..2449
                     /gene="RepA"
                     /label=RepA
     rep_origin      2625..3416
     protein_bind    3491..3512
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3527..3557
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3565..3581
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3589..3605
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3601..4005
                     /codon_start=1
                     /gene="lacZ alpha"
                     /product="beta-galactosidase"
                     /label=lacZ alpha
                     /protein_id="AAD28612.1"
                     /translation="MTMITPSLHACRSTLEDPRPPKAGERERRGSIRLNTCLCTPVYPA
                     DLAAANSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEW
                     RLMRYFLLTHLCGISHRIWCTLSTICSDAA"
     gene            3601..4005
                     /gene="lacZ alpha"
                     /label=lacZ alpha
     misc_feature    3648..3755
                     /gene="lacZ alpha"
                     /label=polylinker
                     /note="polylinker"
     primer_bind     complement(3756..3772)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4246..4350
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4351..5211
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAD28611.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDPGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4351..5211
                     /gene="bla"
                     /label=bla