LOCUS       sequence-429367-        7673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cardiomyocytes-specific plasmid, expressing the coding sequence of 
            mouse Actn2, biotin ligase BioID2, and a HA tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7673)
  AUTHORS   Guo CT, Jardin BD, Lin JS, Ambroise RL, Wang Z, Yang LZ, Mazumdar N,
            Lu FJ, Ma Q, Cao YP, Liu CZ, Li KL, Liu XJ, Lan F, Zhao MM, Xiao H, 
            Dong ED, Pu WT, Guo YX
  TITLE     In vivo proximity proteomics uncovers palmdelphin (PALMD) as a 
            Z-disc-associated mitigator of isoproterenol-induced cardiac injury.
  JOURNAL   Acta Pharmacol Sin. 2024 Jul 23. doi: 10.1038/s41401-024-01348-y.
  PUBMED    39043970
REFERENCE   2  (bases 1 to 7673)
  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"; journalName: "Acta 
            Pharmacol Sin. 2024 Jul 23. doi: 10.1038/s41401-024-01348-y."
COMMENT     Sequence Label: pENAAV-cTNT-actn2-BioID2-HA
FEATURES             Location/Qualifiers
     source          1..7673
                     /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             3722..4417
                     /codon_start=1
                     /product="promiscuous R40G mutant of a biotin protein 
                     ligase from Aquifex aeolicus (Kim et al., 2016)"
                     /label=BioID2
                     /translation="FKNLIWLKEVDSTQERLKEWNVSYGTALVADRQTKGRGGLGRKWL
                     SQEGGLYFSFLLNPKEFENLLQLPLVLGLSVSEALEEITEIPFSLKWPNDVYFQEKKVS
                     GVLCELSKDKLIVGIGINVNQREIPEEIKDRATTLYEITGKDWDRKEVLLKVLKRISEN
                     LKKFKEKSFKEFKGKIESKMLYLGEEVKLLGEGKITGKLVGLSEKGGALILTEEGIKEI
                     LSGEFSLRRS"
     CDS             4418..4444
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4510..4529)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4575..4630
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4723..4852
                     /label=AAV2 ITR
     primer_bind     complement(4871..4888)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4871..4887)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4880..4902)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5029..5484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5116..5135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5326..5347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5510..5614
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5615..5683,5684..6475)
                     /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(5833..5852)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6646..7234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7135..7154
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7522..7543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7558..7563,7564..7581,7582..7588)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7596..7612
                     /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     7601..7623
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7620..7636
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7620..7636
                     /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 gcctcgagaa gcggccgcat gaatcagata gagccaggag tacaatataa
     1021 ttacgtatac gacgaagatg agtatatgat ccaggaggag gagtgggatc gagatctact
     1081 actagatcca gcctgggaga agcagcagag gaagacgttc actgcctggt gcaactcaca
     1141 cctgcggaaa gctggcaccc agatcgagaa catcgaggag gatttcagga acggcctcaa
     1201 gctcatgcta cttctagaag tcatctcagg ggaaaggctg cccaaacctg accggggaaa
     1261 aatgcggttc cacaagattg cgaatgtcaa caaggctctg gattacatag ccagcaaggg
     1321 agtgaagctg gtatctattg gagctgaaga gattgttgac ggcaatgtga agatgacact
     1381 aggtatgata tggacaataa tcctccgctt cgccattcag gacatctcag tggaagaaac
     1441 atctgccaaa gaagggctgc tgctatggtg tcagaggaag acagctccat acagaaatgt
     1501 gaatatccag aacttccaca ccagctggaa agatggcctt ggactctgtg ccctcatcca
     1561 cagacaccgg cccgacctca ttgactactc aaagcttaac aaggatgacc ccataggaaa
     1621 cattaacctg gccatggaaa tagcagagaa gcacttggat atccctaaga tgttggatgc
     1681 tgaagacatt gtcaacactc ccaaacccga tgaaagagcc atcatgactt acgtttcctg
     1741 cttctaccat gctttcgcgg gagcggagca ggccgagaca gcagctaaca gaatatgtaa
     1801 ggttcttgct gtgaatcaag agaatgagag gctgatggaa gaatatgaga ggctagcgag
     1861 tgagctgttg gagtggattc ggcgcacaat cccctggctg gagaacagga cgcctgagaa
     1921 gaccatgcag gccatgcaga agaagctgga ggacttccgg gattaccgtc gtaagcataa
     1981 gccccccaag gtgcaggaga agtgccagct ggagatcaac ttcaacacgc tgcagaccaa
     2041 gcttcggatc agcaaccggc ccgccttcat gccctccgag ggcaagatgg tgtcggacat
     2101 tgctggtgcc tggcagaggt tggagcaggc tgagaagggc tatgaggagt ggctattgaa
     2161 tgagattcga agactggagc gcttggagca cttggctgag aagttcaggc agaaggcctc
     2221 cacgcacgag acatgggctt atggcaaaga acagatcttg ctgcagaagg actacgagtc
     2281 tgcgtccctg acggaagttc gagccctact tcggaagcac gaggcctttg agagcgacct
     2341 ggctgcacac caggaccgcg tggagcagat cgctgccatt gcgcaggagc tcaatgaact
     2401 ggactatcac gatgctgtga acgtcaatga tcggtgccag aaaatttgcg accagtggga
     2461 tagattgggg acgctcaccc agaagaggag ggaggccttg gagagaactg agaagttgct
     2521 ggaaacaatc gaccaactgc acctggagtt tgccaagaga gccgcgccct tcaacaactg
     2581 gatggaggga gccatggagg acctgcagga catgttcatt gtccacagca tcgaggagat
     2641 ccagagtctg atcactgcac atgagcagtt caaggccaca ctgcccgagg ccgacgggga
     2701 gcggcagtcc atcctggcca tccagaacga ggtggagaag gtgatccaga gctacagcat
     2761 ccggatcagc tccagcaacc cctacagcac tgtcaccatg gatgagctgc gcaacaagtg
     2821 ggacaaggtg aagcagctgg tgccggtccg ggatcagtcc ctgcaggagg agctggctcg
     2881 ccaacatgct aatgagcgtc tgcggcgcca gtttgctgct caggccaatg ccattgggcc
     2941 ctggatccag aacaagatgg aggagatcgc tcggagctcc atccagatca ccggggccct
     3001 ggaggaccag atgaaccagc tgaagcagta cgaacacaac atcatcaact acaagaacaa
     3061 catcgacaag ctggagggcg accaccagct catccaggag gccctggtct tcgacaacaa
     3121 gcacacgaac tacaccatgg agcatatccg tgtcggatgg gagttgctgc tgacaactat
     3181 cgccagaacc atcaatgagg tggagaccca gatcctgaca agagatgcca agggcatcac
     3241 ccaggagcag atgaatgagt tcagagcctc cttcaaccac tttgacagga ggaagaacgg
     3301 cctgatggat catgaggatt tcagagcctg cctcatttcc atgggctatg acttgggtga
     3361 agctgaattt gcccgcatta tgactttggt tgaccccaac ggacaaggca cggtcacctt
     3421 ccagtccttc attgacttca tgactagaga gaccgcagac accgacacgg cagagcaggt
     3481 catcgcctcc ttccggattc tggcttctga taagccttac atcttggcag aggagcttcg
     3541 tcgagagctg cctccggatc aggcccagta ctgcatcaag agaatgcccc catactcagg
     3601 cccgggcagt gtccccgggg ccctggacta cactgccttc tcctctgccc tctacgggga
     3661 aagcgacctc actagtggga gcggaggagg gggttcaggc ggaggcggta gcgcccgccg
     3721 gttcaaaaat cttatctggt tgaaggaggt agactctact caagaacggc tgaaggaatg
     3781 gaacgtttcc tatggcaccg cacttgtagc cgataggcaa accaaaggca ggggagggtt
     3841 gggccgaaaa tggctctccc aggaaggggg gctttatttc agcttcttgt tgaacccaaa
     3901 ggagtttgag aacctcctgc aactcccatt ggtgttgggg ctctctgtca gtgaagccct
     3961 tgaagagatc accgagatac cattttcact gaaatggccc aatgatgtgt attttcagga
     4021 gaagaaagta tcaggagtgc tctgcgaact ttcaaaagat aaactgatag ttggcatcgg
     4081 gatcaacgtt aatcagaggg agatcccaga ggaaatcaag gaccgggcca caacccttta
     4141 cgagataact ggcaaggatt gggataggaa agaagtcttg ttgaaggtac tgaagcgcat
     4201 cagcgagaac ctgaagaaat ttaaagagaa gtccttcaag gaattcaagg ggaagatcga
     4261 aagcaagatg ctttatctcg gggaagaagt taaactgctc ggggagggaa aaataaccgg
     4321 gaaactggtc ggcttgagcg aaaaaggggg tgctcttatc ctgaccgaag agggaattaa
     4381 agagatactt tcaggcgagt ttagtttgcg ccgcagttac ccatatgatg tgcccgacta
     4441 tgcttagggt acctctagag tcgacccggg cggcctcgag gacggggtga actacgcctg
     4501 aggatccgat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     4561 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     4621 gtgtctctca ctcggaagca attcgttgat ctgaatttcg accacccata atacccatta
     4681 ccctggtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     4741 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4801 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agccttaatt
     4861 aacctaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     4921 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     4981 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     5041 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5101 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5161 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5221 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     5281 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5341 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     5401 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5461 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5521 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5581 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5641 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5701 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5761 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5821 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5881 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5941 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6001 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6061 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6121 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6181 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6241 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6301 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6361 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6421 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6481 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6541 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6601 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6661 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6721 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6781 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6841 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6901 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6961 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7021 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7081 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     7141 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     7201 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7261 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     7321 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     7381 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     7441 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     7501 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     7561 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     7621 aggaaacagc tatgaccatg attacgccag atttaattaa ggccttaatt agg
//
