LOCUS       sequence-472137-        7303 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to package AAV9:cTNT.iCre, which specifically transduce 
            cardiomyocytes and express Cre recombinase. Only works in vivo..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7303)
  AUTHORS   Lin Z, Zhou P, von Gise A, Gu F, Ma Q, Chen J, Guo H, van Gorp PR, 
            Wang DZ, Pu WT
  TITLE     Pi3kcb links Hippo-YAP and PI3K-AKT signaling pathways to promote 
            cardiomyocyte proliferation and survival.
  JOURNAL   Circ Res. 2015 Jan 2;116(1):35-45. doi: 
            10.1161/CIRCRESAHA.115.304457. Epub 2014 Sep 23.
  PUBMED    25249570
REFERENCE   2  (bases 1 to 7303)
  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: 
            "10.1161/CIRCRESAHA.115.304457"; journalName: "Circ Res"; date: 
            "2015-01-2- 2"; volume: "116"; issue: "1"; pages: "35-45"
COMMENT     Sequence Label: pAAV.cTNT.iCre
FEATURES             Location/Qualifiers
     source          1..7303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          772..904
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     887..906
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        949..967
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             979..999
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    join(2038..2613,2614..2616,2617..2625)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2205..2222)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2432..2451
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2662..4089
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     primer_bind     complement(4140..4159)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4205..4260
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4353..4482
                     /label=AAV2 ITR
     primer_bind     complement(4501..4518)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4501..4517)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4510..4532)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4659..5114
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4746..4765)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4956..4977
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5140..5244
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5245..5313,5314..6105)
                     /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     complement(5463..5482)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6276..6864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6765..6784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7152..7173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7188..7193,7194..7211,7212..7218)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7226..7242
                     /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     7231..7253
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7250..7266
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7250..7266
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctaccag ggtaatgggg
      181 atcctctaga actatagcta gaattcgccc ttacgggccc cccctcgagg tcgggataaa
      241 agcagtctgg gctttcacat gacagcatct ggggctgcgg cagagggtcg ggtccgaagc
      301 gctgccttat cagcgtcccc agccctggga ggtgacagct ggctggcttg tgtcagcccc
      361 tcgggcactc acgtatctcc gtccgacggg tttaaaatag caaaactctg aggccacaca
      421 atagcttggg cttatatggg ctcctgtggg ggaaggggga gcacggaggg ggccggggcc
      481 gctgctgcca aaatagcagc tcacaagtgt tgcattcctc tctgggcgcc gggcacattc
      541 ctgctggctc tgcccgcccc ggggtgggcg ccggggggac cttaaagcct ctgcccccca
      601 aggagccctt cccagacagc cgccggcacc caccgctccg tgggacgatc cccgaagctc
      661 tagagcttta ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac
      721 acaacagtct cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc
      781 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
      841 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
      901 acaggtgtcc actcccagtt caattacagc tcttaaggct agagtactta atacgactca
      961 ctataggcta gcatggtgcc caagaagaag aggaaagtct ccaacctgct gactgtgcac
     1021 caaaacctgc ctgccctccc tgtggatgcc acctctgatg aagtcaggaa gaacctgatg
     1081 gacatgttca gggacaggca ggccttctct gaacacacct ggaagatgct cctgtctgtg
     1141 tgcagatcct gggctgcctg gtgcaagctg aacaacagga aatggttccc tgctgaacct
     1201 gaggatgtga gggactacct cctgtacctg caagccagag gcctggctgt gaagaccatc
     1261 caacagcacc tgggccagct caacatgctg cacaggagat ctggcctgcc tcgcccttct
     1321 gactccaatg ctgtgtccct ggtgatgagg agaatcagaa aggagaatgt ggatgctggg
     1381 gagagagcca agcaggccct ggcctttgaa cgcactgact ttgaccaagt cagatccctg
     1441 atggagaact ctgacagatg ccaggacatc aggaacctgg ccttcctggg cattgcctac
     1501 aacaccctgc tgcgcattgc cgaaattgcc agaatcagag tgaaggacat ctcccgcacc
     1561 gatggtggga gaatgctgat ccacattggc aggaccaaga ccctggtgtc cacagctggt
     1621 gtggagaagg ccctgtccct gggggttacc aagctggtgg agagatggat ctctgtgtct
     1681 ggtgtggctg atgaccccaa caactacctg ttctgccggg tcagaaagaa tggtgtggct
     1741 gccccttctg ccacctccca actgtccacc cgggccctgg aagggatctt tgaggccacc
     1801 caccgcctga tctatggtgc caaggatgac tctgggcaga gatacctggc ctggtctggc
     1861 cactctgcca gagtgggtgc tgccagggac atggccaggg ctggtgtgtc catccctgaa
     1921 atcatgcagg ctggtggctg gaccaatgtg aacattgtga tgaactacat cagaaacctg
     1981 gactctgaga ctggggccat ggtgaggctg ctcgaggatg gggactgagc ggccgcgccc
     2041 ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     2101 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     2161 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     2221 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     2281 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     2341 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     2401 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     2461 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac
     2521 atgctttaca tgtgtttagt cgaggttaaa aaaaacgtct aggccccccg aaccacgggg
     2581 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccccatg gactagtatg
     2641 ggttgctgtt tctccaagac catggtgagc aagggagagg aggtcatcaa agagttcatg
     2701 cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag
     2761 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
     2821 cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac
     2881 gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag
     2941 tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc
     3001 ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac
     3061 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc
     3121 cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac
     3181 tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac
     3241 tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
     3301 cagtacgagc gctccgaggg ccgccaccac ctgttcctgg ggcatggcac cggcagcacc
     3361 ggcagcggca gctccggcac cgcctcctcc gaggacaaca acatggccgt catcaaagag
     3421 ttcatgcgct tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag
     3481 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     3541 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag
     3601 gcgtacgtga agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc
     3661 ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac
     3721 tcctccctgc aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc
     3781 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg
     3841 tacccccgcg acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc
     3901 ggccgctacc tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc
     3961 ggctactact acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc
     4021 gtggaacagt acgagcgctc cgagggccgc caccacctgt tcctgtacgg catggacgag
     4081 ctgtacaagg tcgacccggg cggcctcgag gacggggtga actacgcctg aggatccgat
     4141 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     4201 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     4261 ctcggaagca attcgttgat ctgaatttcg accacccata atacccatta ccctggtaga
     4321 taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac
     4381 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     4441 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agccttaatt aacctaattc
     4501 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4561 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     4621 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     4681 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4741 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4801 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4861 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4921 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4981 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     5041 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5101 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5161 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5221 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5281 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5341 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5401 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5461 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5521 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5581 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5641 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5701 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5761 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5821 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5881 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5941 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6001 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6061 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6121 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6181 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6241 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6301 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6361 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6421 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6481 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6541 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6601 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6661 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6721 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6781 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6841 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6901 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6961 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7021 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7081 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7141 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7201 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7261 tatgaccatg attacgccag atttaattaa ggccttaatt agg
//
