LOCUS       sequence-491760-        7400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for CACNA1C expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7400)
  AUTHORS   Zhonghua Lu Lab
  TITLE     AAVLINK: A Potent DNA-Recombination Method for Large Cargo Delivery 
            in Gene Therapy
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7400)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAVLINK-EFS-HA-5'-CACNA1C
FEATURES             Location/Qualifiers
     source          1..7400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        145..356
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             371..397
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1229..1243)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     repeat_region   4663..4803
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4663..4792
                     /label=AAV2 ITR
     rep_origin      4878..5333
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4965..4984)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5175..5196
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5350..5369)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5469..5491
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5529..5547)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5615..5719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5720..5788,5789..6580)
                     /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(5938..5957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6751..7339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7240..7259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgggcag agcgcacatc gcccacagtc cccgagaagt
      181 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg
      241 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
      301 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagcgta
      361 cgccaccatg tacccttatg acgtacctga ctatgctgga tctgtcaatg agaatacgag
      421 gatgtacatt ccagaggaaa accaccaagg ttccaactat gggagcccac gccccgccca
      481 tgccaacatg aatgccaatg cggcagcggg gctggcccct gagcacatcc ccaccccggg
      541 ggctgccctg tcgtggcagg cggccatcga cgcagcccgg caggctaagc tgatgggcag
      601 cgctggcaat gcgaccatct ccacagtcag ctccacgcag cggaagcggc agcaatatgg
      661 gaaacccaag aagcagggca gcaccacggc cacacgcccg ccccgagccc tgctctgcct
      721 gaccctgaag aaccccatcc ggagggcctg catcagcatt gtcgaatgga aaccatttga
      781 aataattatt ttactgacta tttttgccaa ttgtgtggcc ttagcgatct atattccctt
      841 tccagaagat gattccaacg ccaccaattc caacctggaa cgagtggaat atctctttct
      901 cataattttt acggtggaag cgtttttaaa agtaatcgcc tatggactcc tctttcaccc
      961 caatgcctac ctccgcaacg gctggaacct actagatttt ataattgtgg ttgtggggct
     1021 ttttagtgca attttagaac aagcaaccaa agcagatggg gcaaacgctc tcggagggaa
     1081 aggggccgga tttgatgtga aggcgctgag ggccttccgc gtgctgcgcc ccctgcggct
     1141 ggtgtccgga gtcccaagtc tccaggtggt cctgaattcc atcatcaagg ccatggtccc
     1201 cctgctgcac atcgccctgc ttgtgctgtt tgtcatcatc atctacgcca tcatcggctt
     1261 ggagctcttc atggggaaga tgcacaagac ctgctacaac caggagggca tagcagatgt
     1321 tccagcagaa gatgaccctt ccccttgtgc gctggaaacg ggccacgggc ggcagtgcca
     1381 gaacggcacg gtgtgcaagc ccggctggga tggtcccaag cacggcatca ccaactttga
     1441 caactttgcc ttcgccatgc tcacggtgtt ccagtgcatc accatggagg gctggacgga
     1501 cgtgctgtac tgggtcaatg atgccgtagg aagggactgg ccctggatct attttgttac
     1561 actaatcatc atagggtcat tttttgtact taacttggtt ctcggtgtgc ttagcggaga
     1621 gttttccaaa gagagggaga aggccaaggc ccggggagat ttccagaagc tgcgggagaa
     1681 gcagcagcta gaagaggatc tcaaaggcta cctggattgg atcactcagg ccgaagacat
     1741 cgatcctgag aatgaggacg aaggcatgga tgaggagaag ccccgaaaca tgagcatgcc
     1801 caccagtgag accgagtccg tcaacaccga aaacgtggct ggaggtgaca tcgagggaga
     1861 aaactgtggg gccaggctgg cccaccggat ctccaagtca aagttcagcc gctactggcg
     1921 ccggtggaat cggttctgca gaaggaagtg ccgcgccgca gtcaagtcta atgtcttcta
     1981 ctggctggtg attttcctgg tgttcctcaa cacgctcacc attgcctctg agcactacaa
     2041 ccagcccaac tggctcacag aagtccaaga cacggcaaac aaggccctgc tggccctgtt
     2101 cacggcagag atgctcctga agatgtacag cctgggcctg caggcctact tcgtgtccct
     2161 cttcaaccgc tttgactgct tcgtcgtgtg tggcggcatc ctggagacca tcctggtgga
     2221 gaccaagatc atgtccccac tgggcatctc cgtgctcaga tgcgtccggc tgctgaggat
     2281 tttcaagatc acgaggtact ggaactcctt gagcaacctg gtggcatcct tgctgaactc
     2341 tgtgcgctcc atcgcctccc tgctccttct cctcttcctc ttcatcatca tcttctccct
     2401 cctggggatg cagctctttg gaggaaagtt caactttgat gagatgcaga cccggaggag
     2461 cacattcgat aacttccccc agtccctcct cactgtgttt cagatcctga ccggggagga
     2521 ctggaattcg gtgatgtatg atgggatcat ggcttatggc ggcccctctt ttccagggat
     2581 gttagtctgt atttacttca tcatcctctt catctgtgga aactatatcc tactgaatgt
     2641 gttcttggcc attgctgtgg acaacctggc tgatgctgag agcctcacat ctgcccaaaa
     2701 ggaggaggaa gaggagaagg agagaaagaa gctggccagg actgccagcc cagagaagaa
     2761 acaagagttg gtggagaagc cggcagtggg ggaatccaag gaggagaaga ttgagctgaa
     2821 atccatcacg gctgacggag agtctccacc cgccaccaag atcaacatgg atgacctcca
     2881 gcccaatgaa aatgaggata agagccccta ccccaaccca gaaactacag gagaagagga
     2941 tgaggaggag ccagagatgc ctgtcggccc tcgcccacga ccactctctg agcttcacct
     3001 taaggaaaag gcagtgccca tgccagaagc cagcgcgttt ttcatcttca gctctaacaa
     3061 caggtttcgc ctccagtgcc accgcattgt caatgacacg atcttcacca acctgatcct
     3121 cttcttcatt ctgctcagca gcatttccct ggctgctgag gacccggtcc agcacacctc
     3181 cttcaggaac catattctgt tttattttga tattgttttt accaccattt tcaccattga
     3241 aattgctctg aagatgactg cttatggggc tttcttgcac aagggttctt tctgccggaa
     3301 ctacttcaac atcctggacc tgctggtggt cagcgtgtcc ctcatctcct ttggcatcca
     3361 gtccagtgca atcaatgtcg tgaagatctt gcgagtcctg cgagtactca ggcccctgag
     3421 ggccatcaac agggccaagg ggctaaagca tgtggttcag tgtgtgtttg tcgccatccg
     3481 gaccatcggg aacatcgtga ttgtcaccac cctgctgcag ttcatgtttg cctgcattgg
     3541 ggtccagctc ttcaagggaa agctgtacac ctgttcagac agttccaagc agacagaggc
     3601 ggaatgcaag ggcaactaca tcacgtacaa agacggggag gttgaccacc ccatcatcca
     3661 accccgcagc tgggagaaca gcaagtttga ctttgacaat gttctggcag ccatgatggc
     3721 cctcttcacc gtctccacct tcgaagggtg gccagagctg ctgtaccgct ccatcgactc
     3781 ccacacggaa gacaagggcc ccatctacaa ctaccgtgtg gagatctcca tcttcttcat
     3841 catctacatc atcatcatcg ccttcttcat gatgaacatc ttcgtgggct tcgtcatcgt
     3901 cacctttcag gagcaggggg agcaggagta caagaactgt gagctggaca agaaccagcg
     3961 acagtgcgtg gaatacgccc tcaaggcccg gcccctgcgg aggtacatcc ccaagaacca
     4021 gcaccagtac aaagtgtggt acgtggtcaa ctccacctac ttcgagtacc tgatgttcgt
     4081 cctcatcctg ctcaacacca tctgcctggc catgcagcac tacggccaga gctgcctgtt
     4141 caaaatcgcc atgaacatcc tcaacatgct cttcactggc ctcttcaccg tggagatgat
     4201 cctgaagctc attgccttca aacccaagca ctatttctgt gatgcatgga atacatttga
     4261 cgccttgatt gttgtgggta gcattgttga tatagcaatc accgaggtaa acccagctga
     4321 acatacccaa tgctctccct ctatgaacgc agaggaaaac tcccgcatct ccatcacctt
     4381 cttccgcctg ttccgggtca tgcgtctggt gaagctgctg agccgtgggg agggcatccg
     4441 gacgctgctg tggaccttca tcaagtcctt ccaggccctg ccctatgtgg ccctcctgat
     4501 cgtgatgctg ttcttcatct acgcggtgat cgggatgcag gtaagtatca aggttacaag
     4561 acaggtttaa ggagaccaat aagctattat tcgtatagga tactttatac gaagttatta
     4621 agttatggag accggtgtgg cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     4681 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4741 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     4801 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     4861 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4921 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     4981 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5041 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     5101 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5161 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     5221 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     5281 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     5341 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     5401 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5461 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5521 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5581 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5641 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5701 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5761 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5821 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5881 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5941 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6001 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6061 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6121 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6181 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6241 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6301 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6361 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6421 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6481 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6541 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6601 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6661 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6721 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6781 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6841 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6901 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6961 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7021 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7081 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7141 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7201 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7261 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7321 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7381 ctggcctttt gctcacatgt
//
