LOCUS       sequence-7767-ex        7369 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 7369)
  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 7369)
  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: pNrl-Cre
FEATURES             Location/Qualifiers
     source          1..7369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      3229..3238
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3235..4263
                     /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             4273..4302
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4393..4412)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4458..4513
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4512..4531)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4872..4888)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4872..4888)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4885..4907)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4896..4912
                     /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(4920..4926,4927..4944,4945..4950))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4965..4986
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5045..5240
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5091..5226
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5246..5380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5283..5302)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5337..5356
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5448..5465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5619..6207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5699..5718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6378..7169,7170..7238))
                     /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     7001..7020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7239..7343)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacttac atgtacatac ttcataatgg tctagaaggg cttattcaaa tttggcgttt
       61 actcagagtc ctcggggctc ttatccggga tgtggtgatg atgtgaggag aggcaactct
      121 caaactagga ccatcactcg gggcttgtgt atcagtagtg aggtccagca atggaaggga
      181 aatcccggga aaaaagtcta tgtgatccct aggagcaggt ctgagctctt tgtgcagaaa
      241 atttctggat acctggagta tgattctcag ggaccagggg tctgggtcag agaagggtgg
      301 ctgtctcaca tcatggggta cattcagcta tagaaagacc agcatgaacc aggcagtggt
      361 acctggcctg aaccaggact ggctcagctt gcttgagctt atttggcccg actcccttca
      421 ctccttagca gcttagctct cccacctctc tcctgagtcc tccagtaaac aagaggatca
      481 ggacagatat gacagctgcc tggtggggca agtgtctggt caggtcaaca gcaggcatga
      541 agggggctac tacactaaat gctgccaacc tttttccttt agggaagttc taggatctct
      601 ttcccggata tccactgtaa aacctcattc cactggcatg tatccctatt cctttcagca
      661 tccctattgc ccttttgtgg atggttcctg gccttacatt ttccgcttct tttaagaaaa
      721 taagcaacag gtcttggtga agagtaacat ttccattcga atagtgactc tatttcataa
      781 accaaacgaa acgaaacgaa acgaaacgaa acgaaacaaa acaaaacaaa acaaaacacc
      841 aagagacctg ggcatgggag agttgtgata tctgacctaa gccttctaat gtatgtagag
      901 gaagcttgga ttccaaatgt cctccaaagc tgggcatggt ggtgcgtgcc tttacgccct
      961 gtactaggaa ggcacaagca ggtagatcta tgggttcaag gtcaatctgg tctacaaaga
     1021 gagttctagg acggctaggg ctacacagag aaaccctgtt ttgaaaacca aaccaaacca
     1081 aaccaatgtc ctccaaaggc atttgagttc aaaacttgtc attattggaa ggtggtgaaa
     1141 ttttcaagag gtggggccct gtgggaagtc ttctggtctt tgggttttcc ctgaaggaag
     1201 caaaggccta acccctccct ggtcatcggt tggtaccctt gagggaagca ggagtcctat
     1261 ccacttccct tcccttttct tttttttttt ttttgtccca gtgataaggt gagtagcttt
     1321 cagttgtgtc ctcgccataa cacttgactc actacaggca atacatgatg ggaccaactg
     1381 ctcactggct aggctctcca aggcagcaaa ccaaagcaaa gcgctttccg attaagtcaa
     1441 ttacttcagg tatgtattgg agcaattatg ttagtgttag tcggtttttg ttattgtgac
     1501 aaagtgcccg agaaagatca aaggagggga gccttctttg ggctaatggc ttcagtttca
     1561 gctgatggtt gtgggtgtcc tattgctttg gtccaaaatc taacatcacg atggaaaatg
     1621 tatagtgaag aagtaaagag agaaaagaga gaggaagagg gggaagtgga ggtgggggag
     1681 aaggagggat ggagggacac agacaaaatg caccaatgaa ctaaggagaa gatacaccat
     1741 tcaaacgcat gttttttcta gttaggccaa ttcatactga cagccaattc agtatggatt
     1801 cttcagctgg ttagttatgg ataagtttaa cctctaataa tatctcagtg aaaactagct
     1861 tttaaaatat accggaacca tagcagtgat agaaaactaa tgtagggtgg acacacacac
     1921 acacacacac acacaaacac acactttgag actgggttga aggtaggcat ctgtggacca
     1981 gtgggtctca aacattagtg tgtttcagag ctgttggaag ggtttattaa acagcagctg
     2041 tctataacct acttactgag tttctggttc tgtgagtttg aggtaaagca aaagaatttg
     2101 tttcttcaag ttcccaaggg gtgatttctt aagactgtgg aagccattgt tccaacaggc
     2161 agggaatacc ctttaacttc tggagtttct tttatcacag cacctattta aacagcttca
     2221 ggcaggttta aacatctcct tggctaagtc ctactatccc tgcttcggcc cctactaggg
     2281 ggtgacagct ctaagaggcg ttaggcgatt aggctgaaaa tgtaggtcac accccagccg
     2341 tctgggaatg agcgagagag gagaagaggg actgagacac cgcacgggga aaaggtgaga
     2401 ggaagctgag ggctcatgag ggcattcaca gaagtgtggt ggtgattgga tttcagcttc
     2461 ttttagtctc cagatgaaag accagagttg ggatctccgc tttttctatc tcttctctcc
     2521 agctgggctg cttgtcttcg ggatgggcta ttaagagcga gtttcttcat ggaagctcat
     2581 ggcttatctt tctaatacag tgactctgtg aggaataaga cgagtcaggg tgaaggtagg
     2641 gcaggggctg aaatgtgagg atgcttgaaa agaagcagca gccccaacaa gctggctgag
     2701 gatgggctgg gaacgcgtat atgtctcctt gacacggagt cttaaaagag gttttgggta
     2761 gtatgtctgg actcttacca ctctaagctt tgagtgattg ggctgagaat tgtgtttcca
     2821 tggaggggtg gtgtttgggg tcaacatggc tctctatatt ggaaaccaag ctcacaacat
     2881 aagcaaggtc tgatatagga agcttctctg gggaagatgc cgtttgtttt tttccttttt
     2941 ggagaactga tagcacctgt ctttcactgg cttctgagtc cctgccccag gccaatgaga
     3001 tatgcaaata agcccttcat cttattggat gaggctcgag gaccacagat gatctcaggg
     3061 aaccagtcct ttaagagtgt cctctgggct ctgtgcccac agcccccttg ttctgaatac
     3121 agggacgaca ccagcccctg ctctatggag tatttagtct ccagggaagc tgtgcctttc
     3181 tggttctgac agtgactacg tcatctctgc cattacgaat tctgagccgc caccatggcc
     3241 aatttactga ccgtacacca aaatttgcct gcattaccgg tcgatgcaac gagtgatgag
     3301 gttcgcaaga acctgatgga catgttcagg gatcgccagg cgttttctga gcatacctgg
     3361 aaaatgcttc tgtccgtttg ccggtcgtgg gcggcatggt gcaagttgaa taaccggaaa
     3421 tggtttcccg cagaacctga agatgttcgc gattatcttc tatatcttca ggcgcgcggt
     3481 ctggcagtaa aaactatcca gcaacatttg ggccagctaa acatgcttca tcgtcggtcc
     3541 gggctgccac gaccaagtga cagcaatgct gtttcactgg ttatgcggcg gatccgaaaa
     3601 gaaaacgttg atgccggtga acgtgcaaaa caggctctag cgttcgaacg cactgatttc
     3661 gaccaggttc gttcactcat ggaaaatagc gatcgctgcc aggatatacg taatctggca
     3721 tttctgggga ttgcttataa caccctgtta cgtatagccg aaattgccag gatcagggtt
     3781 aaagatatct cacgtactga cggtgggaga atgttaatcc atattggcag aacgaaaacg
     3841 ctggttagca ccgcaggtgt agagaaggca cttagcctgg gggtaactaa actggtcgag
     3901 cgatggattt ccgtctctgg tgtagctgat gatccgaata actacctgtt ttgccgggtc
     3961 agaaaaaatg gtgttgccgc gccatctgcc accagccagc tatcaactcg cgccctggaa
     4021 gggatttttg aagcaactca tcgattgatt tacggcgcta aggatgactc tggtcagaga
     4081 tacctggcct ggtctggaca cagtgcccgt gtcggagccg cgcgagatat ggcccgcgct
     4141 ggagtttcaa taccggagat catgcaagct ggtggctgga ccaatgtaaa tattgtcatg
     4201 aactatatcc gtaacctgga tagtgaaaca ggggcaatgg tgcgcctgct ggaagatggc
     4261 gatggaccgg tggaacaaaa acttatttct gaagaagatc tgtgatagcg gccgcactcc
     4321 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     4381 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     4441 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     4501 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     4561 gtatttggtt tagagtttgg caacatatgc catatgctgg ctgccatgaa caaaggtggc
     4621 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     4681 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     4741 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     4801 tcccagtcat agctgtccct cttctcttat gaagatccct cgacctgcag cccaagcttg
     4861 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4921 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4981 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     5041 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5101 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5161 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5221 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     5281 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5341 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     5401 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5461 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5521 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5581 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5641 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5701 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5761 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5821 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5881 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5941 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6001 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6061 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6121 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6181 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6241 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6301 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6361 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6421 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6481 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6541 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6601 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6661 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6721 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6781 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6841 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6901 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6961 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7021 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7081 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7141 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7201 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7261 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7321 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
