LOCUS       sequence-7766-ex        6454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6454)
  AUTHORS   Matsuda T, Cepko CL
  TITLE     Controlled expression of transgenes introduced by in vivo 
            electroporation.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 Jan 16;104(3):1027-32. doi: 
            10.1073/pnas.0610155104
  PUBMED    17209010
REFERENCE   2  (bases 1 to 6454)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1073/pnas"; 
            journalName: "Proc Natl Acad Sci U S A"; date: "2007-01-16- 16"; 
            volume: "104"; issue: "3"; pages: "1027-32"
COMMENT     Sequence Label: pRho-Cre
FEATURES             Location/Qualifiers
     source          1..6454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2314..2323
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2320..3348
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             3358..3387
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3478..3497)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3543..3598
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3597..3616)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3957..3973)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3957..3973)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3970..3992)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3981..3997
                     /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(join(4005..4011,4012..4029,4030..4035))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4050..4071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4130..4325
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4176..4311
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4331..4465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4368..4387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4422..4441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4533..4550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4704..5292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4784..4803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5463..6254,6255..6323))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6086..6105
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6324..6428)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agccagcttc cacgttgtca gaagcgttag cgggtaacaa
       61 gcaaggccat gagtttgagg gtcgggggga cagcctggcc agggcagggc tgcgtgctct
      121 ggtcttgacc cagggagtcc atcgtctgtc cctctgtctg gccaccaggg gcgtgttaga
      181 atcaaactct tcaccttgac ctctttagcg atggacctaa tcacagggcc aagcaccttg
      241 agcagctgcg ccccacccac ccgccacacc tgcctgcccc ttgctccttc cccccagagc
      301 acttaccacc aggctcccca cacaagtcct gatgctgccc gctgcccggg acagcagggc
      361 tctggagccc aggtgggctg cagcgaaggg agcacgtgtc tgagcagaga ggtgtagaca
      421 tcccaggtgg taagagggac agcacaaggg agcctcagcg atggaggtct cagacactgg
      481 gatgcttcct ggccaccctt gcgttcccca cctcggggcc tttgcactgg ccgctctctc
      541 tccttggaca gctccacccc ttagtgctcc ttccatcctc gagttctcag agaggccttc
      601 ctgggcggct ccaggcaact gtgtgtcggg aggacgtcac catctgaaca gtccctgtgg
      661 tcgtttgttt catggtttgt gggggcccct ccccgccccc ccagaagaag gacacgtcct
      721 acaggcaggg gcttggtcct tcttgtccac tgtgtccccc agaactgagc gtcttgtgta
      781 gaacacagtc ggtgccagat aaaactcaca gctggactgt ggtcattgag gaccccgaag
      841 gccaggctgc ccgcgacccc acttcactcc taggaggggt cttcacgatc ctattgtaga
      901 tgggggccct tgttcgcacc ctgagctcgt gctgctgttc tgggatacgc catcacggct
      961 tgccgatcgc acggcacccg aggcccacgg ccctaaccca gcctgacccc agtcctggcc
     1021 tcggcccgtg tgctgttccc agggcccacc tcccctccat atgggagtgc ccgggaccac
     1081 acctccttgc tcaccgcagg gatctggtgc ccggccaagg cttggccccg ttcacctcct
     1141 ggagacgagg ccttgcttca ggagccctaa gagcttctgt cccctccccg ctttggttcc
     1201 ctctgccctg gtcccttcct ccaccatcgc tcccttcagc tcatccttga gggcttttga
     1261 gccaccttcc tctccccaag atctgcccaa gttcccctgg ataaagccct ggcactaagc
     1321 cccaccccag ctgaccctag cccacgaggg cccacatccc agatgggcct cgggccttca
     1381 ccgggcctct gtcttcagtc tatgctgact tccccacttg gcaacctggc aaagtgcccc
     1441 cctccctcaa cacattgtcc ccctggacgt ctctgcagat aggtggcctc cccgccccgg
     1501 gtccagagcg ggtcttccca cacggaagtc ctcctcagcc cataagctcc tccttcctgg
     1561 agcctcgggt gccccagtgg cctgcacata aatgtcggtt tgaacaagtg agtgagtgaa
     1621 ctccagttca agggaacaaa tgctttttga gctcctgggc aacgggggct tgtgtccggt
     1681 ccccggagcg ccccgctgca cttgtgaagc tcacgacctg gccggtgggc actgtcagtg
     1741 tgtggtgact ggataagccc agaggtgacc caggtgggga agggacagct caggggctga
     1801 gtgaggggat gaccgaagga ccagcgctct gccctccagg agctcgtggg ccaggcggga
     1861 ggaaggcaaa atttgcttgc cctgagccag agggaagtgg gcctagagct aggggtccca
     1921 atgccaggcc aggcccatca gctgagatgc cagaggggga caggggagtt gggggcaggc
     1981 ctctgctctt tcccagggtc cccagcacgc cccgccttct ccccgaccac agccttctac
     2041 ctggagtcac cttggcccca cctggaagcc aattaagccc ctcgttgcag cagtgaggat
     2101 taatatgatt aataacgccc ccaatctccg aggtgctgat tcagccggga gcttagggag
     2161 gggaggtcac ttcataaggg cttggggggg gagttggagc cacgagtcgt ccagccggag
     2221 ccccgtgtgg ctgagctccg gcctcagaag catccccggg ttcgcgccgc cggggatcct
     2281 ctagagtcga gctcaagctt cgaattctga gccgccacca tggccaattt actgaccgta
     2341 caccaaaatt tgcctgcatt accggtcgat gcaacgagtg atgaggttcg caagaacctg
     2401 atggacatgt tcagggatcg ccaggcgttt tctgagcata cctggaaaat gcttctgtcc
     2461 gtttgccggt cgtgggcggc atggtgcaag ttgaataacc ggaaatggtt tcccgcagaa
     2521 cctgaagatg ttcgcgatta tcttctatat cttcaggcgc gcggtctggc agtaaaaact
     2581 atccagcaac atttgggcca gctaaacatg cttcatcgtc ggtccgggct gccacgacca
     2641 agtgacagca atgctgtttc actggttatg cggcggatcc gaaaagaaaa cgttgatgcc
     2701 ggtgaacgtg caaaacaggc tctagcgttc gaacgcactg atttcgacca ggttcgttca
     2761 ctcatggaaa atagcgatcg ctgccaggat atacgtaatc tggcatttct ggggattgct
     2821 tataacaccc tgttacgtat agccgaaatt gccaggatca gggttaaaga tatctcacgt
     2881 actgacggtg ggagaatgtt aatccatatt ggcagaacga aaacgctggt tagcaccgca
     2941 ggtgtagaga aggcacttag cctgggggta actaaactgg tcgagcgatg gatttccgtc
     3001 tctggtgtag ctgatgatcc gaataactac ctgttttgcc gggtcagaaa aaatggtgtt
     3061 gccgcgccat ctgccaccag ccagctatca actcgcgccc tggaagggat ttttgaagca
     3121 actcatcgat tgatttacgg cgctaaggat gactctggtc agagatacct ggcctggtct
     3181 ggacacagtg cccgtgtcgg agccgcgcga gatatggccc gcgctggagt ttcaataccg
     3241 gagatcatgc aagctggtgg ctggaccaat gtaaatattg tcatgaacta tatccgtaac
     3301 ctggatagtg aaacaggggc aatggtgcgc ctgctggaag atggcgatgg accggtggaa
     3361 caaaaactta tttctgaaga agatctgtga tagcggccgc actcctcagg tgcaggctgc
     3421 ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct
     3481 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     3541 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     3601 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     3661 tttggcaaca tatgccatat gctggctgcc atgaacaaag gtggctataa agaggtcatc
     3721 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     3781 gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt
     3841 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     3901 tccctcttct cttatgaaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
     3961 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4021 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4081 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     4141 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4201 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4261 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4321 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4381 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4441 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     4501 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4561 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4621 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4681 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4741 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4801 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4861 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     4921 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4981 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5041 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5101 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5161 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5221 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5281 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5341 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5401 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5461 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5521 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5581 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5641 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5701 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5761 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5821 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5881 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5941 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6001 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6061 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6121 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6181 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6241 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6301 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6361 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6421 gttccgcgca catttccccg aaaagtgcca cctg
//
