LOCUS       Exported                5653 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBLCAT3T-p1, complete sequence.
ACCESSION   AM697711
VERSION     .
KEYWORDS    ampR gene; beta-lactamase; cat gene; chloramphenicol
            acetyltra...
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5653)
  AUTHORS   Luckow B, Schutz G.
  TITLE     CAT constructions with multiple unique restriction sites for the 
            functional analysis of eukaryotic promoters and regulatory elements
  JOURNAL   Nucleic Acids Res. 15 (13), 5490 (1987)
  PUBMED    3037497
REFERENCE   2  (bases 1 to 5653)
  AUTHORS   Lobo SM, Hernandez N.
  TITLE     A 7 bp mutation converts a human RNA polymerase II snRNA promoter 
            into an RNA polymerase III promoter
  JOURNAL   Cell 58 (1), 55-67 (1989)
  PUBMED    2752422
REFERENCE   3  (bases 1 to 5653)
  AUTHORS   Principaud E, Spohr G.
  TITLE     Xenopus laevis c-myc I and II genes: molecular structure and 
            developmental expression
  JOURNAL   Nucleic Acids Res. 19 (11), 3081-3088 (1991)
  PUBMED    2057364
REFERENCE   4  (bases 1 to 5653)
  AUTHORS   Bernhardt K.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-APR-2007) Bernhardt K., The Gurdon Institute, 
            University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, 
            UNITED KINGDOM
REFERENCE   5  (bases 1 to 5653)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5653
                     /organism="Cloning vector pBLCAT3T-p1"
                     /focus
                     /mol_type="other DNA"
                     /note="this cloning vector is based on pBLCAT3, Accession 
                     Number X64409, used for promoter reporter assays"
                     /db_xref="taxon:439690"
     source          755..1753
                     /organism="Mus musculus"
                     /chromosome="6"
                     /map="6F2"
                     /mol_type="other DNA"
                     /db_xref="taxon:10090"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      440..453
                     /regulatory_class="terminator"
                     /note="this is a nonfunctional RNA polymerase II terminator
                     (3 box), termination depends on the presence of an 
                     appropriate snRNA promoter upstream such as in the human U1
                     and U2 snRNA genes"
                     /citation="[2]"
                     /citation="[3]"
     regulatory      474..481
                     /regulatory_class="terminator"
                     /note="this is a functional RNA polymerase III terminator 
                     from the human U6 snRNA gene"
                     /citation="[2]"
                     /citation="[3]"
     regulatory      755..1753
                     /regulatory_class="promoter"
                     /gene="Nanog"
                     /note="999 bp of the mouse Nanog promoter -1000 to -2 with 
                     respect to the transcriptional start site (-1190 to -192 
                     with respect to the Nanog start codon)"
     gene            755..1753
                     /gene="Nanog"
                     /label=Nanog
     CDS             1797..2456
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /function="reporter gene"
                     /label=cat
                     /citation="[1]"
                     /protein_id="CAM92322.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            1797..2456
                     /gene="cat"
                     /label=cat
     intron          2546..3159
                     /note="SV40 small T intron"
                     /citation="[1]"
     intron          2622..2687
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2817..2837
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3156..3396
                     /label=SV40 poly(A)
                     /note="SV40 poly(A)"
                     /citation="[1]"
     polyA_signal    3262..3396
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3432..3448)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3456..3472
                     /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)."
     promoter        complement(3480..3510)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3525..3546
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3834..4422)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4593..5453)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=ampR
                     /citation="[1]"
                     /protein_id="CAM92323.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4593..5453)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(5454..5558)
                     /gene="bla"
                     /label=AmpR promoter