LOCUS pAAV.cTNT.iCre 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
//
