LOCUS       sequence-458120-        7177 bp    DNA     circular SYN 15-APR-2025
DEFINITION  cardiomyocytes-specific plasmid with Tnnt2 promotor, expressing the 
            coding sequence of mouse Atp2a2 and a HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7177)
  TITLE     Myocardial infarction creates a critical time window for AAV gene 
            therapy
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7177)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-Tnnt2 -Atp2a2-HA
FEATURES             Location/Qualifiers
     source          1..7177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     CDS             3908..3934
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4014..4033)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4079..4134
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4227..4356
                     /label=AAV2 ITR
     primer_bind     complement(4375..4392)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4375..4391)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4384..4406)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4533..4988
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4620..4639)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4830..4851
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5014..5118
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5119..5187,5188..5979)
                     /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(5337..5356)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6150..6738
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6639..6658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7026..7047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7062..7067,7068..7085,7086..7092)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7100..7116
                     /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     7105..7127
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7124..7140
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7124..7140
                     /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 actatagctc agtccattag gagccagtag cctggaagat gtctttaccc
      241 ccagcatcag ttcaagtgga gcagcacata actcttgccc tctgccttcc aagattctgg
      301 tgctgagact tatggagtgt cttggaggtt gccttctgcc ccccaaccct gctcccagct
      361 ggccctccca ggcctgggtt gctggcctct gctttatcag gattctcaag agggacagct
      421 ggtttatgtt gcatgactgt tccctgcata tctgctctgg ttttaaatag cttatctgag
      481 cagctggagg accacatggg cttatatggc gtggggtaca tgttcctgta gccttgtccc
      541 tggcacctgc caaaatagca gccaacaccc cccaccccca ccgccatccc cctgccccac
      601 ccgtcccctg tcgcacattc ctccctccgc agggctggct caccaggccc cagcccacat
      661 gcctgcttaa agccctctcc atcctctgcc tcacccagtc cccgctgaga ctgagcagac
      721 gcctccagga tctgtcggca gagctcctcg agaagcggcc gcactactcc gcatggagaa
      781 cgctcacaca aagaccgtgg aggaggtgct gggccacttc ggggtcaacg agagcacggg
      841 gctgagcttg gagcaggtca agaagctcaa ggagagatgg ggctccaacg aattgccggc
      901 tgaagaagga aaaaccttgc tggaacttgt gattgagcag tttgaagact tactagttag
      961 aattttactg ctggcagcat gtatatcttt cgttttggct tggttcgagg aaggggaaga
     1021 aacgattaca gcctttgtag agccgtttgt aattctgctt atcttggtag ccaatgcaat
     1081 cgtgggtgtg tggcaggaaa gaaatgctga aaatgcaata gaagctctta aggaatatga
     1141 gcctgaaatg ggcaaagtgt atcgacagga cagaaagagt gtgcaacgaa ttaaagctaa
     1201 agacatagtt cctggtgata tagtggaaat tgctgttggt gacaaagttc ctgctgatat
     1261 tagattgaca tccatcaagt ctacaactct aagagtcgac cagtcaattc ttacaggtga
     1321 atctgtctcc gtcatcaagc atactgaccc tgtccctgac ccccgagctg ttaatcaaga
     1381 caaaaagaac atgctctttt ctggtacaaa cattgctgct gggaaagcta tgggagtggt
     1441 ggtggcaact ggagttaata ctgagatcgg caagatccgg gatgaaatgg tggcaacaga
     1501 acaggagaga acacccctac agcagaagct agacgagttt ggggagcagc tttccaaagt
     1561 tatctccctc atttgcattg cagtctggat catcaacatt gggcatttca atgacccagt
     1621 tcatggtggc tcctggatca ggggtgccat ctactacttt aagattgccg tggccctggc
     1681 tgttgccgca atccctgagg gtctgcctgc tgtcatcacc acctgcttag ctcttggaac
     1741 tcgtaggatg gcaaagaaaa atgctatcgt tcgaagtctg ccttctgtgg agacccttgg
     1801 ttgtacttct gttatctgct cagataagac aggcacactt accacaaacc agatgtccgt
     1861 gtgcaggatg ttcattctgg acaaagtaga aggtgacact tgttccctta atgagttcag
     1921 cataactgga tccacatatg caccaattgg agaagtgcaa aaggatgata agccagtgaa
     1981 gtgccatcag tatgacgggc ttgtagagtt agccaccatc tgtgctctgt gtaatgactc
     2041 tgctttggat tataatgagg caaagggtgt gtatgagaaa gttggagaag ctaccgagac
     2101 tgctctcacg tgcctggtgg agaagatgaa tgtatttgat actgagctga aggggctttc
     2161 taaaatagag cgtgcaaacg cctgcaactc ggtcataaag cagctgatga agaaggagtt
     2221 cactctggag ttttcacggg atagaaaatc aatgtccgtc tattgtaccc caaacaagcc
     2281 aagccggaca tccatgagca agatgtttgt gaagggggct ccagaaggtg tcatcgatag
     2341 gtgcacccac atccgagttg gaagtaccaa ggtccccatg actcctggtg tcaaacagaa
     2401 gattatgtct gtcattcggg agtggggcag tggcagcgac acgctacggt gcctggctct
     2461 ggccactcat gacaacccac tgaagagaga ggagatgcac ctggaagact ctgctaactt
     2521 catcaaatac gagaccaacc tgactttcgt cggctgtgtg ggcatgctgg atcctcccag
     2581 gattgaagta gcctcttctg tgaagctgtg ccggcaagca ggcatccggg tcatcatgat
     2641 cactggagac aacaaaggca ccgctgtggc catctgtcgc cgcattggca tctttgggca
     2701 ggatgaggat gtgacatcaa aggcttttac agggcgagag tttgatgaat taagcccttc
     2761 agcccagaga gatgcctgct taaatgcccg ctgttttgct cgagttgaac cttcccacaa
     2821 gtctaagatt gttgagttcc ttcagtcctt tgatgagatc acagctatga ctggtgatgg
     2881 tgtgaatgat gctcctgctc tgaagaaatc tgaaatcggg attgccatgg gctcagggac
     2941 tgcagtggct aagactgctt ctgagatggt cctggcagat gacaacttct ccaccattgt
     3001 ggctgctgtt gaggaggggc gagccatcta caacaacatg aagcagttca tccgctacct
     3061 catctcatcc aacgtggggg aagtggtctg tatcttcctg acggcagccc ttgggtttcc
     3121 tgaggcttta attcctgtcc agttactctg ggtcaatctg gtgacagatg gtctgcctgc
     3181 cactgcgctg gggttcaatc ctccagacct ggacatcatg aacaaacccc cccggaaccc
     3241 aaaagaacca ctgatcagcg ggtggctctt tttccgttac ctggctattg gctgttatgt
     3301 tggcgctgcc accgtgggtg ctgctgcatg gtggttcatc gctgctgacg gcggtccaag
     3361 agtctccttc taccagctga gtcatttcct acagtgtaag gaggacaacc cagacttcga
     3421 tggagtggat tgtgcaatct ttgagtcccc atatccaatg acaatggcac tttctgttct
     3481 agtaaccata gagatgtgta atgccctcaa cagcttgtct gaaaaccagt ctttgctgag
     3541 gatgcccccc tgggagaata tctggctcgt gggctccatc tgcttgtcca tgtcacttca
     3601 cttcttgatc ctctacgtgg aacctttgcc gctcattttc cagatcacac cgctgaatct
     3661 gacccagtgg ctgatggtgc tgaaaatctc cttgcctgtg atcctcatgg atgagacgct
     3721 caagtttgtg gcccgaaact acctggaaca acccggtaaa gagtgtgtgc agcctgccac
     3781 caaatcttcc tgctccctgt cggcatgcac cgatggcatt tcctggccgt ttgtgctgct
     3841 cattatgccc ctggtggtct gggtctacag cacagacact aactttagcg atatgttctg
     3901 gtctggttac ccatacgatg ttcctgacta tgcgtgataa taagctcgcg tggtacctct
     3961 agagtcgacc cgggcggcct cgaggacggg gtgaactacg cctgaggatc cgatcttttt
     4021 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     4081 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     4141 agcaattcgt tgatctgaat ttcgaccacc cataataccc attaccctgg tagataagta
     4201 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     4261 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4321 tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc
     4381 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4441 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4501 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     4561 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4621 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4681 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4741 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4801 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4861 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4921 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     4981 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5041 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5101 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5161 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5221 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5281 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5341 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5401 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5461 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5521 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5581 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5641 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5701 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5761 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5821 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5881 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5941 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6001 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6061 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6121 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6181 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6241 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6301 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6361 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6421 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6481 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6541 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6601 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6661 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6721 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6781 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6841 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6901 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     6961 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     7021 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     7081 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     7141 catgattacg ccagatttaa ttaaggcctt aattagg
//
