LOCUS       sequence-454205-        5661 bp    DNA     circular SYN 15-APR-2025
DEFINITION  cardiomyocytes-specific plasmid with Tnnt2 promotor, expressing the 
            coding sequence of luciferase and a HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5661)
  TITLE     Myocardial infarction creates a critical time window for AAV gene 
            therapy
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5661)
  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            "Unpublished"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-HA-Luciferase
FEATURES             Location/Qualifiers
     source          1..5661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     CDS             760..786
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             787..2436
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(850..868)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2498..2517)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2563..2618
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   2711..2840
                     /label=AAV2 ITR
     primer_bind     complement(2859..2876)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2859..2875)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2868..2890)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3017..3472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3104..3123)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3314..3335
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3498..3602
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3603..3671,3672..4463)
                     /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(3821..3840)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4634..5222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5123..5142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5510..5531
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5546..5551,5552..5569,5570..5576)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5584..5600
                     /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     5589..5611
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5608..5624
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5608..5624
                     /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 agaagcatgt acccatatga tgtgcccgac
      781 tatgctgaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat
      841 ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca
      901 attgctttta cagatgcaca tatcgaggtg gacatcactt acgctgagta cttcgaaatg
      961 tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc
     1021 gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga
     1081 gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgggc
     1141 atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg
     1201 caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg attctaaaac ggattaccag
     1261 ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac
     1321 gattttgtgc cagagtcctt cgatagggac aagacaattg cactgatcat gaactcctct
     1381 ggatctactg gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg cgtgagattc
     1441 tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt
     1501 gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga
     1561 tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttctgaggag ccttcaggat
     1621 tacaagattc aaagtgcgct gctggtgcca accctattct ccttcttcgc caaaagcact
     1681 ctgattgaca aatacgattt atctaattta cacgaaattg cttctggtgg cgctcccctc
     1741 tctaaggaag tcggggaagc ggttgccaag aggttccatc tgccaggtat caggcaagga
     1801 tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg
     1861 ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg
     1921 aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc
     1981 ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat
     2041 tctggagaca tagcttactg ggacgaagac gaacacttct tcatcgttga ccgcctgaag
     2101 tctctgatta agtacaaagg ctatcaggtg gctcccgctg aattggaatc catcttgctc
     2161 caacacccca acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa
     2221 cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg
     2281 gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg
     2341 gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc
     2401 ataaaggcca agaagggcgg aaagatcgcc gtgtaagtac ctctagagtc gacccgggcg
     2461 gcctcgagga cggggtgaac tacgcctgag gatccgatct ttttccctct gccaaaaatt
     2521 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     2581 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaagcaat tcgttgatct
     2641 gaatttcgac cacccataat acccattacc ctggtagata agtagcatgg cgggttaatc
     2701 attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     2761 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     2821 gtgagcgagc gagcgcgcag ccttaattaa cctaattcac tggccgtcgt tttacaacgt
     2881 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     2941 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3001 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3061 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3121 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3181 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3241 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3301 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3361 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3421 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     3481 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3541 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3601 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3661 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3721 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3781 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3841 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3901 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3961 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4021 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4081 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4141 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4201 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4261 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4321 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4381 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4441 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4501 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4561 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4621 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4681 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4741 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4801 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4861 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4921 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4981 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5041 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5101 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5161 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5221 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5281 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5341 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5401 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     5461 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     5521 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     5581 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccagat
     5641 ttaattaagg ccttaattag g
//
