LOCUS       sequence-491938-        6771 bp    DNA     circular SYN 05-NOV-2025
DEFINITION  3' AAVLINK plasmid for MYO5C expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6771)
  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  JOURNAL   Unpublished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 6771)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAVLINKv1-pCALM1-Cre-3'-MYO5C-FLAG
FEATURES             Location/Qualifiers
     source          1..6771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             277..1308
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     primer_bind     1464..1483
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3513..3578
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3643..3663)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     rep_origin      4249..4704
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4336..4355)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4546..4567
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4721..4740)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4840..4862
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4900..4918)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4986..5090
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5091..5159,5160..5951)
                     /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(5309..5328)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6122..6710
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6611..6630
                     /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 gcgtcgctct gcgcgaggcg gctccgccgc ggcggaggga
      181 tacggcgcac catatatata tcgcggggcg cagactcgcg ctccggcagt ggtgctggga
      241 gtgtcgtgga cgccgtgccg ttactcgtac gccaccatgt ccaatttact gaccgtacac
      301 caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg
      361 gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct tctgtccgtt
      421 tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc cgcagaacct
      481 gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt aaaaaccatc
      541 cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc acgaccaagt
      601 gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt
      661 gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc
      721 atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg gattgcttat
      781 aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat ctcacgtact
      841 gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag caccgcaggt
      901 gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat ttccgtctct
      961 ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa tggtgttgcc
     1021 gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt tgaagcaact
     1081 catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc ctggtctgga
     1141 cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc aataccggag
     1201 atcatgcaag ctggtggctg gaccaatgta aatattgtaa tgaactatat ccgtaacctg
     1261 gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattaggt aagtatcaag
     1321 gttacaagac aggtttaagg agaccaataa ctagtataac ttcgtatagg atactttata
     1381 gcaatttatg aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     1441 tattggtctt actgacatcc actttgcctt tctctccaca ggtcaagaca atctctgagt
     1501 tcgaaaaaga gatagaacta cttcaggcac agaagataga tgtggagaaa catgtgcagt
     1561 cacaaaaacg ggaaatgaga gaaaagatgt cagagatcac caaacaactt ctcgaaagct
     1621 atgacattga agatgtaaga agcaggctct ctgtggaaga tctggaacat ttaaatgagg
     1681 atggagaact ttggtttgct tatgaaggac taaagaaagc aacacgtgtt ttggagagcc
     1741 atttccagtc tcagaaggat tgctatgaaa aggagattga agctttgaac ttcaaagtgg
     1801 tgcatctcag tcaagaaatc aaccacctgc agaagttatt cagagaagaa aatgacatca
     1861 atgaaagcat ccgtcatgaa gttaccaggc taacatcaga gaacatgatg atcccagact
     1921 ttaaacagca aatttcagaa ttggagaaac agaagcaaga tcttgaaatc cgcctgaatg
     1981 aacaagctga gaaaatgaaa ggaaaactag aagaattgtc taatcagtta catcgcagtc
     2041 aagaggagga aggaacacaa agaaaggcct tggaagccca aaatgaaata cataccaaag
     2101 agaaggagaa gctgattgat aagattcaag aaatgcagga ggccagtgac cacttgaaga
     2161 aacaatttga aactgaaagt gaagtcaagt gtaacttccg gcaggaagca tcccggctca
     2221 ctttggaaaa cagggatctt gaagaagaat tagacatgaa agacagagtg attaaaaagc
     2281 tacaagatca agtcaagaca ctaagcaaga caattggaaa agccaatgat gtgcactcgt
     2341 cctcaggacc caaggagtac cttggaatgc tgcaatacaa gagagaagac gaggccaagc
     2401 tcattcagaa cctcattctt gacttgaagc cccgtggcgt ggtggtgaac atgatccccg
     2461 ggctgccggc tcatatcctg ttcatgtgtg tgcgctacgc agactctctg aatgatgcca
     2521 acatgctgaa gtccctcatg aacagcacca ttaatggcat caagcaggtg gttaaggaac
     2581 atttagaaga ctttgaaatg ctgtcctttt ggctttccaa cacttgtcat tttctcaatt
     2641 gcctgaagca gtacagcgga gaagaggaat tcatgaagca taatagtcca cagcagaata
     2701 agaattgctt gaacaatttt gacctttcag aatacagaca gattctcagt gatgtggcta
     2761 tacgaatata tcatcaattt attatcataa tggaaaagaa tatccaaccg ataatagttc
     2821 cgggaatgct ggagtatgag agcctgcagg gcatttccgg cctgaagccc acaggcttcc
     2881 ggaagcgctc ctctagcata gacgacacgg acggctacac catgacctcc gtcctgcaac
     2941 agctgagcta cttttacacc accatgtgcc agaacggcct ggaccccgag cttgtgaggc
     3001 aggcggtgaa gcagctcttc ttcttgatcg gggcggtcac gctgaacagc ctcttcctgc
     3061 gcaaggacat gtgctcctgc agaaaaggga tgcagatcag gtgcaatatc agctacttag
     3121 aagaatggct taaagataag aacttgcaga acagcttagc aaaggaaact ttggagcccc
     3181 tctctcaggc agcctggttg cttcaggtca agaagaccac agacagtgat gccaaggaga
     3241 tctacgaacg ctgcacctca ctgtctgctg tgcagatcat aaagatcctt aattcataca
     3301 cacctataga tgactttgag aagagagtga ctccatcctt tgttcgcaaa gtacaggctc
     3361 tcctaaatag ccgggaggat tcatcacagc tgatgttgga taccaaatat ctctttcaag
     3421 tcacatttcc ttttaccccc tctccacatg ctctggaaat gattcagatc cccagcagtt
     3481 tcaagctagg ctttctgaat aggttatgta cagactacaa agaccatgac ggtgattata
     3541 aagatcatga catcgactac aaggatgacg atgacaagta actcgagata atcaacctct
     3601 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3661 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3721 tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca tcgccgcctg
     3781 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgtttat
     3841 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatc tagctttatt tgtgaaattt
     3901 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     3961 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagatctg
     4021 gagaccggtg tggcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     4081 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccggcc
     4141 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4201 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4261 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4321 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4381 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4441 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4501 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4561 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     4621 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4681 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4741 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4801 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4861 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4921 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4981 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5041 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5101 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5161 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5221 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5281 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5341 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5401 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5461 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5521 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5581 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5641 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5701 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5761 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5821 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5881 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5941 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6001 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6061 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6121 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6181 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6241 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6301 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6361 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6421 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6481 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6541 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6601 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6661 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6721 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
