LOCUS       Exported                7302 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Used to package AAV9:cTNT.iCre, which specifically transduce 
            cardiomyocytes and express Cre recombinase. Only works in vivo..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.cTNT.iCre
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7302)
  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 7302)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7302
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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    2038..2624
                     /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             2661..4088
                     /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(4139..4158)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4204..4259
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4352..4481
                     /label=AAV2 ITR
     primer_bind     complement(4500..4517)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4500..4516)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4509..4531)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4658..5113
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4745..4764)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4955..4976
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5139..5243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5244..6104
                     /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(5462..5481)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6275..6863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6764..6783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7017..7034
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7151..7172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7187..7217
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7225..7241
                     /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     7230..7252
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7249..7265
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7249..7265
                     /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 aacatagtga 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 ctcggtgcac
     2521 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     2581 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccccatgg actagtatgg
     2641 gttgctgttt ctccaagacc atggtgagca agggagagga ggtcatcaaa gagttcatgc
     2701 gcttcaaggt gcgcatggag ggctccatga acggccacga gttcgagatc gagggcgagg
     2761 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc
     2821 ccctgccctt cgcctgggac atcctgtccc cccagttcat gtacggctcc aaggcgtacg
     2881 tgaagcaccc cgccgacatc cccgattaca agaagctgtc cttccccgag ggcttcaagt
     2941 gggagcgcgt gatgaacttc gaggacggcg gtctggtgac cgtgacccag gactcctccc
     3001 tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc ccccccgacg
     3061 gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc
     3121 gcgacggcgt gctgaagggc gagatccacc aggccctgaa gctgaaggac ggcggccact
     3181 acctggtgga gttcaagacc atctacatgg ccaagaagcc cgtgcaactg cccggctact
     3241 actacgtgga caccaagctg gacatcacct cccacaacga ggactacacc atcgtggaac
     3301 agtacgagcg ctccgagggc cgccaccacc tgttcctggg gcatggcacc ggcagcaccg
     3361 gcagcggcag ctccggcacc gcctcctccg aggacaacaa catggccgtc atcaaagagt
     3421 tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg
     3481 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     3541 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg
     3601 cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct
     3661 tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact
     3721 cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc
     3781 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt
     3841 acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg
     3901 gccgctacct ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg
     3961 gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg
     4021 tggaacagta cgagcgctcc gagggccgcc accacctgtt cctgtacggc atggacgagc
     4081 tgtacaaggt cgacccgggc ggcctcgagg acggggtgaa ctacgcctga ggatccgatc
     4141 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     4201 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     4261 tcggaagcaa ttcgttgatc tgaatttcga ccacccataa tacccattac cctggtagat
     4321 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     4381 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4441 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gccttaatta acctaattca
     4501 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4561 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4621 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     4681 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4741 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4801 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4861 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4921 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4981 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5041 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5101 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5161 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5221 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5281 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5341 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5401 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5461 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5521 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5581 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5641 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5701 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5761 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5821 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5881 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5941 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6001 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6061 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6121 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6181 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6241 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6301 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6361 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6421 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6481 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6541 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6601 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6661 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6721 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6781 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6841 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6901 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6961 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     7021 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     7081 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     7141 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     7201 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     7261 atgaccatga ttacgccaga tttaattaag gccttaatta gg
//
