LOCUS       sequence-429377-        7027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cardiomyocytes-specific plasmid, expressing the coding sequence of 
            mouse Nexn and GFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7027)
  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 7027)
  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: pAAV-cTNT-NEXN-GFP
FEATURES             Location/Qualifiers
     source          1..7027
                     /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             join(3071..3073,3074..3076,3077..3790)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3071..3073,3074..3076,3077..3787)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3116..3137)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3377..3396)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3724..3745
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3864..3883)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3929..3984
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4077..4206
                     /label=AAV2 ITR
     primer_bind     complement(4225..4242)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4225..4241)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4234..4256)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4383..4838
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4470..4489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4680..4701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4864..4968
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4969..5037,5038..5829)
                     /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(5187..5206)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6000..6588
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6489..6508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6876..6897
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6912..6917,6918..6935,6936..6942)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6950..6966
                     /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     6955..6977
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6974..6990
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6974..6990
                     /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 gcggccgcac tactccgcgg actactacta gtatgaatga
     1021 cgtttcgcaa aaggcagaga ttctgctttc ctcatgtaaa cctgtcccta agtcctatgt
     1081 gccaaagctt ggcaagggag atgtaaagga caagtttgaa gccatgcaga gggccaggga
     1141 agaaagaaat caaaggcgat ctagagacga aaagcaaaga agaaaagaac aatatattag
     1201 agagagagaa tggaacagga gaaagcagga gattaaagaa atgcttgctt ctgatgatga
     1261 ggaagaaagt tctcctaaaa tagaaaaggc ttatgtccca aagctaacag ggacagttaa
     1321 aggtaaattc gatgaaatgg agaaacacag acaggaagaa caaagaaaga gaacagaaga
     1381 agaaagaaaa cgcagaattg agcaggatct gttagaaaag aggaaaatac agcgcgaatt
     1441 agcaaaaaga gcggagcaga ttgaggacat aaacaatacg ggaaccgagt ccgcatcaga
     1501 ggaaggagat gactcactac ttataacagt ggtacctgcc aaatcataca aaacacctgg
     1561 aaaaacaaag gatcctgagg acctggacag agaagaagga aatgggagaa ccaaccatga
     1621 agaagataaa atgagatatg aagaagagtg ccgagttctc aaagaagcaa aatgtctttc
     1681 attagtcatg gatgatgaaa cagaggccaa aaaagaatca cattttcctg gaaaattgaa
     1741 atcaactttt gaagaactgg aacgacaaag acaagaaaat cgcaagaagc aagcggaaga
     1801 ggaagcgaga cgacgcttag aagaagagag acgttccttt gaggaggcga gaaggcacat
     1861 ggtaaatgag gaggatgaaa accaagatag agaaacagtt tttaaagagt accgcccagg
     1921 aaaactcaaa ctcagttttg aggaaataga aaggcaaagg agagaagacg aaaaacgaaa
     1981 agcagaggag gaggcccgaa ggagaataga ggaagagaag gcggcatttg ctgaagcaag
     2041 gagaagcatg gtattagatg atgattcccc agagatttat aaaacagtgt ctcaagaatc
     2101 tcttactcct ggaaaactag aaattaattt tgaacaatta ttaagacaaa aaatggaaga
     2161 agaaagacga cgaacagagg aagaacgcag gcataaacta gaaatggaga aacaggaatt
     2221 tgaacaactg agacaagaaa tgggaaagga agaagaagaa aatgaaagct ttgggttgag
     2281 tagagaatat gaagaattaa ttaaattaaa aaggagtggc tcaattcaag ctaaaaatct
     2341 aaaaagcaag tttgaaaaaa ttggacaatt gtctgaaaaa gaagtacaga aaaagataga
     2401 agaagagcga gcaaaaaggc gagcaattga ccttgaaatc aaagagcgag aagcagaaaa
     2461 cttccatgag gacgatgatg ttgatgtaag gcctgcaaag aagagtgagt ctccttttac
     2521 tcataaggtg aacatgaaag ccaggtttga acaaatggca aaggcaagag aagaggaaga
     2581 acaaagaaga attgaagaac aaaagttact acgaatgcag tttgaacaga aagaaatcga
     2641 tgcggcatta cagaagaaaa gagaagatga ggaggaagaa gaaggtagca ttgttaatgg
     2701 ctccactact gaagatgaag agcaaaccag atcaggagct ccatggttca aaaagccttt
     2761 aagaaatacc tcagttgtag acagtgagcc agtcagattt actgttaaag taacaggaga
     2821 acccaagccg gaaattacat ggtggtttga aggagaaata ctgcaggatg gagaagacta
     2881 tcagtacatc gaaagaggtg aaacttactg cctgtattta ccggaaacct tcccagaaga
     2941 tggaggagag tacatgtgta aggcagtcaa caataaaggc tcagcagcga gcacctgcat
     3001 tcttaccatt gaaatggatg actacggtgg cggtggctcg ggcggtggtg ggtcgggcgg
     3061 tggtgggtcg atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     3121 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     3181 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3241 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     3301 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     3361 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     3421 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     3481 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     3541 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     3601 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     3661 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3721 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3781 gtacaagtaa taagctcgcg tggtacctct agagtcgacc cgggcggcct cgaggacggg
     3841 gtgaactacg cctgaggatc cgatcttttt ccctctgcca aaaattatgg ggacatcatg
     3901 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     3961 tgttggaatt ttttgtgtct ctcactcgga agcaattcgt tgatctgaat ttcgaccacc
     4021 cataataccc attaccctgg tagataagta gcatggcggg ttaatcatta actacaagga
     4081 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4141 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4201 gcgcagcctt aattaaccta attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     4261 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     4321 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4381 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4441 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4501 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4561 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4621 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4681 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4741 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4801 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     4861 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4921 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4981 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5041 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5101 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5161 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5221 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5281 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5341 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5401 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5461 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5521 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5581 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5641 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5701 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5761 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5821 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5881 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5941 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6001 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6061 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6121 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6181 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6241 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6301 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6361 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6421 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6481 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6541 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6601 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6661 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6721 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6781 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     6841 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     6901 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     6961 taacaatttc acacaggaaa cagctatgac catgattacg ccagatttaa ttaaggcctt
     7021 aattagg
//
