LOCUS       sequence-491869-        6967 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  3' AAVLINK plasmid for ATP10A expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6967)
  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  JOURNAL   Unpublished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 6967)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 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            "Unpublished"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'-ATP10A-FLAG
FEATURES             Location/Qualifiers
     source          1..6967
                     /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"
     CDS             3687..3752
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3817..3837)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4230..4370
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4230..4359
                     /label=AAV2 ITR
     rep_origin      4445..4900
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4532..4551)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4742..4763
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4917..4936)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5036..5058
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5096..5114)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5182..5286
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5287..5355,5356..6147)
                     /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(5505..5524)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6318..6906
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6807..6826
                     /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 gggggccgac tcagtggtca
     1501 tggatctcct gcagccctgc tcttcagttg acgccagagg gaggcatcaa aaaaagattc
     1561 ggagcaaaac tcagaattac ctcaacgtgt atgcggcgga aggcctgcgc accttgtgca
     1621 tcgccaagag agttctgagt aaagaagagt atgcctgctg gttgcaaagc cacctagaag
     1681 ccgaatcctc cctggaaaac agcgaggagc tcctcttcca gtctgccatt cgcctggaga
     1741 ccaacctgca cttgttaggt gccactggga ttgaagaccg cctgcaggac ggagtccctg
     1801 aaactatttc taaattgcgt caagcgggcc tgcagatttg ggttctcact ggtgacaaac
     1861 aagaaacagc tgtcaacatt gcatatgcct gcaaactgct ggaccacgac gaggaggtca
     1921 tcaccctgaa tgccacctcc caggaggcgt gtgcagccct gctagaccag tgcctatgct
     1981 acgtgcagtc cagaggcctc cagagagccc ctgagaagac caagggcaaa gtgagcatga
     2041 ggttctcctc tctctgccca ccctccacgt ccactgcctc tggccgcaga cccagcctcg
     2101 tgatcgatgg gagaagcctg gcctacgctc tcgagaaaaa cctggaggac aaattcctct
     2161 tccttgccaa gcagtgccgc tccgtcctct gctgtcggtc gacgcctctg cagaagagca
     2221 tggtggtgaa gctggtgcgg agcaagctca aggccatgac cctggccata ggtgatggag
     2281 ccaatgatgt cagcatgatc caggtggcag atgtgggtgt gggaatctcc ggccaggagg
     2341 gtatgcaggc agtgatggcc agcgactttg cagtgccgaa attccgatac ctggagaggc
     2401 tcttgattct tcacgggcat tggtgctact cccgacttgc caacatggtg ctgtacttct
     2461 tctacaaaaa cacaatgttc gtgggcctcc tgttttggtt ccagtttttc tgtggcttct
     2521 ctgcatctac catgattgac cagtggtatc taatcttctt taatctgctc ttctcgtcac
     2581 ttcccccgct cgtgactggg gtgctggaca gggatgtgcc agccaatgtg ctgctgacca
     2641 acccgcagct ctacaagagt ggccagaaca tggaggaata ccggccacga acgttctggt
     2701 ttaacatggc cgacgccgcc ttccagagcc tggtttgctt ttccattcct tacctggcct
     2761 actatgactc gaacgtggac ctgtttacct gggggacccc tattgtgaca atcgcgctgc
     2821 tcactttcct gctccacctg ggcattgaaa ccaaaacctg gacctggctc aactggataa
     2881 cgtgtggctt cagtgtcctt ttgtttttca ccgtggcttt gatttacaat gcgtcttgtg
     2941 ccacgtgcta tcctccgtcc aacccttact ggactatgca agccttactg ggtgacccag
     3001 tgttttactt gacttgcctg atgacgcctg tcgctgcact gctgcccaga ttgtttttca
     3061 gatccctcca ggggagggtt ttccccacac aacttcagct ggcacgtcag ttgaccagga
     3121 agtcccccag gagatgcagt gctcccaaag agacctttgc tcagggacgc ctcccgaagg
     3181 actcgggaac cgagcactca tcagggagga cagtcaagac ctctgtgccc ctgtcccagc
     3241 cttcttggca cacacagcag ccggtctgct ccctggaggc cagcggggag cccagcacag
     3301 tggacatgag catgccagtg agggagcaca ccctgctgga ggggctgagc gcaccggccc
     3361 ccatgtcctc tgcgccaggg gaggctgtcc tgaggagtcc aggagggtgt cctgaggagt
     3421 ccaaggtgag agctgccagc accggcaggg tgacccccct gtcttccctc ttcagcctgc
     3481 ctaccttcag cttactcaac tggatttcct cctggtcgct ggtcagcagg ctggggagtg
     3541 tcttacagtt ctcccggacg gagcagcttg cagatggaca agcgggacgt ggacttcctg
     3601 tccagcccca ctcaggccga tcaggacttc aagggccaga ccacagacta cttataggag
     3661 catcttcaag gcggtcacag tgtacagact acaaagacca tgacggtgat tataaagatc
     3721 atgacatcga ctacaaggat gacgatgaca agtaactcga gataatcaac ctctggatta
     3781 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3841 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3901 ctccttgtat aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc
     3961 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt ttatttgtga
     4021 aatttgtgat gctattgctt tatttgtaac catctagctt tatttgtgaa atttgtgatg
     4081 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     4141 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaaga tctggagacc
     4201 ggtgtggcac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc
     4261 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     4321 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     4381 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     4441 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4501 accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc
     4561 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4621 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     4681 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4741 agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat
     4801 ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa
     4861 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     4921 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     4981 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     5041 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     5101 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5161 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5221 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5281 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5341 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5401 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5461 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5521 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5581 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5641 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5701 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5761 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5821 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5881 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5941 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6001 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6061 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6121 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6181 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6241 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6301 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6361 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6421 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6481 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6541 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6601 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6661 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6721 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6781 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6841 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6901 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6961 cacatgt
//
