LOCUS       Exported                9479 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPRCG, complete sequence.
ACCESSION   AY569778
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9479)
  AUTHORS   Wu G, Wang Y, Zhu Z.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAR-2004) Fish Genetics, Institute of Hydrobiology, 
            Chinese Academy of Science, 7# South Road of East Lake, Wuhan, Hubei
            430072, Peoples Republic of China
REFERENCE   2  (bases 1 to 9479)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9479
                     /organism="Cloning vector pPRCG"
                     /mol_type="other DNA"
                     /db_xref="taxon:268732"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    419..452
                     /label=LoxP RE
                     /note="LoxP RE"
     protein_bind    419..452
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      459..2499
                     /regulatory_class="promoter"
                     /note="salmon protamine promoter"
     regulatory      823..832
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      2506..2526
                     /regulatory_class="other"
                     /note="SV40 T-antigen NLS"
     CDS             2506..2526
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2536..5187
                     /codon_start=1
                     /product="T7 RNA polymerase"
                     /label=T7 RNA polymerase
                     /protein_id="AAS78492.1"
                     /translation="MNTINIAKNDFSDIELAAIPFNTLADHYGERLAREQLALEHESYE
                     MGEARFRKMFERQLKAGEVADNAAAKPLITTLLPKMIARINDWFEEVKAKRGKRPTAFQ
                     FLQEIKPEAVAYITIKTTLACLTSADNTTVQAVASAIGRAIEDEARFGRIRDLEAKHFK
                     KNVEEQLNKRVGHVYKKAFMQVVEADMLSKGLLGGEAWSSWHKEDSIHVGVRCIEMLIE
                     STGMVSLHRQNAGVVGQDSETIELAPEYAEAIATRAGALAGISPMFQPCVVPPKPWTGI
                     TGGGYWANGRRPLALVRTHSKKALMRYEDVYMPEVYKAINIAQNTAWKINKKVLAVANV
                     ITKWKHCPVEDIPAIEREELPMKPEDIDMNPEALTAWKRAAAAVYRKDKARKSRRISLE
                     FMLEQANKFANHKAIWFPYNMDWRGRVYAVSMFNPQGNDMTKGLLTLAKGKPIGKEGYY
                     WLKIHGANCAGVDKVPFPERIKFIEENHENIMACAKSPLENTWWAEQDSPFCFLAFCFE
                     YAGVQHHGLSYNCSLPLAFDGSCSGIQHFSAMLRDEVGGRAVNLLPSETVQDIYGIVAK
                     KVNEILQADAINGTDNEVVTVTDENTGEISEKVKLGTKALAGQWLAYGVTRSVTKRSVM
                     TLAYGSKEFGFRQQVLEDTIQPAIDSGKGLMFTQPNQAAGYMAKLIWESVSVTVVAAVE
                     AMNWLKSAAKLLAAEVKDKKTGEILRKRCAVHWVTPDGFPVWQEYKKPIQTRLNLMFLG
                     QFRLQPTINTNKDSEIDAHKQESGIAPNFVHSQDGSHLRKTVVWAHEKYGIESFALIHD
                     SFGTIPADAANLFKAVRETMVDTYESCDVLADFYDQFADQLHESQLDKMPALPAKGNLN
                     LRDILESDFAFA"
     polyA_signal    5366..5487
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      5397..5447
                     /regulatory_class="terminator"
                     /note="SV40 polyA"
     regulatory      5537..6126
                     /regulatory_class="promoter"
                     /note="CMV promoter"
     enhancer        5597..5900
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5901..6104
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             6212..6931
                     /codon_start=1
                     /product="eGFP"
                     /label=eGFP
                     /protein_id="AAS78493.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    7054..7175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      7085..7135
                     /regulatory_class="terminator"
                     /note="SV40 polyA"
     misc_feature    7188..7221
                     /label=LoxP LE
                     /note="LoxP LE"
     protein_bind    7188..7221
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(7258..7274)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7282..7298
                     /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(7306..7336)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7351..7372
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7660..8248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      7660
                     /label=Col E1
                     /note="Col E1"
     CDS             complement(8419..9279)
                     /codon_start=1
                     /product="Amp"
                     /label=Amp
                     /protein_id="AAS78494.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9280..9384)
                     /gene="bla"
                     /label=AmpR promoter