LOCUS       sequence-476857-        6336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for SMARCA2 expression .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6336)
  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  JOURNAL   Unpublished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 6336)
  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            "Unpublished"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: pAAVLINKv1-pCALM1-Cre-3' SMARCA2
FEATURES             Location/Qualifiers
     source          1..6336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1030..1159
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1221..1809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1301..1320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1980..2771,2772..2840))
                     /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     2603..2622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2841..2945)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3013..3031
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3069..3091)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3191..3210
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3227..3682)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3364..3385)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3576..3595
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3757..3897
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3768..3897
                     /label=AAV2 ITR
     primer_bind     4290..4310
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4375..4440)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(join(6188..6336,1..883))
                     /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"
ORIGIN
        1 ctccgacacg ggcactgtgt ccagaccagg ccaggtatct ctgaccagag tcatccttag
       61 cgccgtaaat caatcgatga gttgcttcaa aaatcccttc cagggcgcga gttgatagct
      121 ggctggtggc agatggcgcg gcaacaccat tttttctgac ccggcaaaac aggtagttat
      181 tcggatcatc agctacacca gagacggaaa tccatcgctc gaccagttta gttaccccca
      241 ggctaagtgc cttctctaca cctgcggtgc taaccagcgt tttcgttctg ccaatatgga
      301 ttaacattct cccaccgtca gtacgtgaga tatctttaac cctgatcctg gcaatttcgg
      361 ctatacgtaa cagggtgtta taagcaatcc ccagaaatgc cagattacgt atatcctggc
      421 agcgatcgct attttccatg agtgaacgaa cctggtcgaa atcagtgcgt tcgaacgcta
      481 gagcctgttt tgcacgttca ccggcatcaa cgttttcttt tcggatccgc cgcataacca
      541 gtgaaacagc attgctgtca cttggtcgtg gcagcccgga ccgacgatga agcatgttta
      601 gctggcccaa atgttgctgg atggttttta ctgccagacc gcgcgcctga agatatagaa
      661 gataatcgcg aacatcttca ggttctgcgg gaaaccattt ccggttattc aacttgcacc
      721 atgccgccca cgaccggcaa acggacagaa gcattttcca ggtatgctca gaaaacgcct
      781 ggcgatccct gaacatgtcc atcaggttct tgcgaacctc atcactcgtt gcatcgaccg
      841 gtaatgcagg caaattttgg tgtacggtca gtaaattgga catggtggcg tacgagtaac
      901 ggcacggcgt ccacgacact cccagcacca ctgccggagc gcgagtctgc gccccgcgat
      961 atatatatgg tgcgccgtat ccctccgccg cggcggagcc gcctcgcgca gagcgacgcg
     1021 tgcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     1081 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg cggcctcagt gagcgagcga
     1141 gcgcgcagct gcctgcagga catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1201 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1261 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1321 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1381 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1441 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1501 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1561 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1621 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1681 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1741 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1801 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1861 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1921 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1981 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2041 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2101 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2161 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2221 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2281 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2341 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2401 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2461 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2521 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2581 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2641 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2701 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2761 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2821 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2881 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2941 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     3001 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     3061 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     3121 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     3181 cttaactatg cggcatcaga gcagattgta ctgagagtgc accataaaat tgtaaacgtt
     3241 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3301 accgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagataga gttgagtgtt
     3361 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3421 aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc aagttttttg
     3481 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3541 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3601 gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3661 aatgcgccgc tacagggcgc gtactatggt tgctttgacg tatgcggtgt gaaataccgc
     3721 acagatgcgt aaggagaaaa taccgcatca ggcgcccctg caggcagctg cgcgctcgct
     3781 cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct
     3841 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
     3901 gccgctcggt ccgcacgtgc cacaccggtc tccagatctt taaaaaacct cccacatctc
     3961 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4021 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc tagatggtta
     4081 caaataaagc aatagcatca caaatttcac aaataaacac cacggaattg tcagtgccca
     4141 acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc gccgtggcaa
     4201 gaactaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat
     4261 gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa
     4321 taccagtcaa tctttcacaa attttgtaat ccagaggttg attatctcga gttacttgtc
     4381 atcgtcatcc ttgtagtcga tgtcatgatc tttataatca ccgtcatggt ctttgtagtc
     4441 tgtacactca tcatccgtcc cacttccttc tgactgttca cgttcatcct gctcctcatc
     4501 gctgtcaaaa tcgctcacta caggtttggc ttttcctcga tttggccttt tcttgccttt
     4561 ccctttgtcc cggcctttgt catctttttt attgagctta attttcacct tgactgattt
     4621 tgcctcggac tctgactctt cttcatcttc ctcttcctcc tcttcattgc tttcatcctc
     4681 actctcttcc tctttggcaa ttttctgccg ggcactctta aacactgact gtaagacgat
     4741 ggagtcttca tagatctggg atccctccag gttgaacgtc tgagcgttgt gacagagaag
     4801 catgacatcc ttctccaggt cgcctaggct ccggtactta tgattacgaa tcctttcctt
     4861 tatttttttg aaatccactg gcttcctaat taattcatag tattctggta attctttcct
     4921 tgaaggtaac tgaatgaaga cttcactgag ctgtcgccct gaactatctt tgtagtttat
     4981 cacagtatcg atgatagcgt tcatctgctt tgtcagtttg gggggatttg gtgacagttt
     5041 ctcagcggga gggcggcctc ttctcttctt agctttttcc acatcttctt ttgcaggatc
     5101 tttatccaca tttcttcgtc tttttcgctt cttaagccgt acttcctctt ccatttcctc
     5161 caaattgccg tcttcgatgg cccttagcca ctgcttctcc gtgagggcgt cactgtagtc
     5221 cacgtcacgg cgctggcggg accccctccc aaatattttc tcctcctctt cttcacaggt
     5281 gagcctttct acttcagcgt catccttaat gatccaggag ggcagctcat cctcctccat
     5341 taaacggggc ttccgtttcg ggttccgggc atcttccctc cgccggtcca tgtccatccg
     5401 cataaaaagg tcaaattctt cttctcgtcg agcaatcatt tggttcagag tctcatcgtc
     5461 cggtacttca tcttcttcct cattttcctc ctcatgctcc aagatggcct gcaggaatgc
     5521 cctccgctcg tggcttgaag acttttggtc aaacatgccc gcctggatca ctttctgatc
     5581 cacgttcagc ttgtattttg cggccgcgag gatcttttcc tccacgctgt tcacggtaca
     5641 gagcctcagt acccggacct cgttctgctg cccgatgcgg tgagctcggt cttgggcctg
     5701 cagatcctga tgaggattcc agtcgctgtc aaagatgacc actgtatcag ctgcctgaag
     5761 atttaagccc aggccaccag ctcttgtgct cagcaagaaa atgaaatact gggatccagg
     5821 ttcattgaat ttcttcagca aagcagcacg atcttcagac ttggtggtgc catcaaggcg
     5881 taggtaaagg aagttccgaa aagcaaaata atcctccatg atggtcatga gagatgtcat
     5941 ctggcagaaa agcagcactc ggtgattagt cgctctcaat tttggcagaa tacgatcaag
     6001 cagctcaaac ttccctgtgg agagaaaggc aaagtggatg tcagtaagac caataggtgc
     6061 ctatcagaaa cgcaagagtc ttctctgtct cgacaagccc agtttcataa attgctataa
     6121 agtatcctat acgaagttat actagttatt ggtctcctta aacctgtctt gtaaccttga
     6181 tacttaccta atcgccatct tccagcaggc gcaccattgc ccctgtttca ctatccaggt
     6241 tacggatata gttcattaca atatttacat tggtccagcc accagcttgc atgatctccg
     6301 gtattgaaac tccagcgcgg gccatatctc gcgcgg
//
