LOCUS       sequence-491421-        6763 bp    DNA     circular SYN 03-NOV-2025
DEFINITION  3' AAVLINK plasmid for SMARCA4 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6763)
  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  JOURNAL   Unpublished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 6763)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 3, 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            "Unpublished"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'-SMARCA4-FLAG
FEATURES             Location/Qualifiers
     source          1..6763
                     /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             3483..3548
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3613..3633)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4026..4166
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4026..4155
                     /label=AAV2 ITR
     rep_origin      4241..4696
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4328..4347)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4538..4559
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4713..4732)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4832..4854
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4892..4910)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4978..5082
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5083..5151,5152..5943)
                     /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(5301..5320)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6114..6702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6603..6622
                     /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 gggcgtgctg ctgactgatg
     1501 gctccgagaa ggacaagaag ggcaaaggcg gcaccaagac cctgatgaac accatcatgc
     1561 agctgcggaa gatctgcaac cacccctaca tgttccagca catcgaggag tccttttccg
     1621 agcacttggg gttcactggc ggcattgtcc aagggctgga cctgtaccga gcctcgggta
     1681 aatttgagct tcttgataga attcttccca aactccgagc aaccaaccac aaagtgctgc
     1741 tgttctgcca aatgacctcc ctcatgacca tcatggaaga ttactttgcg tatcgcggct
     1801 ttaaatacct caggcttgat ggaaccacga aggcggagga ccggggcatg ctgctgaaaa
     1861 ccttcaacga gcccggctct gagtacttca tcttcctgct cagcacccgg gctggggggc
     1921 tcggcctgaa cctccagtcg gcagacactg tgatcatttt tgacagcgac tggaatcctc
     1981 accaggacct gcaagcgcag gaccgagccc accgcatcgg gcagcagaac gaggtgcgtg
     2041 tgctccgcct ctgcaccgtc aacagcgtgg aggagaagat cctagctgca gccaagtaca
     2101 agctcaacgt ggaccagaag gtgatccagg ccggcatgtt cgaccagaag tcctccagcc
     2161 atgagcggcg cgccttcctg caggccatcc tggagcacga ggagcaggat gagagcagac
     2221 actgcagcac gggcagcggc agtgccagct tcgcccacac tgcccctccg ccagcgggcg
     2281 tcaaccccga cttggaggag ccacctctaa aggaggaaga cgaggtgccc gacgacgaga
     2341 ccgtcaacca gatgatcgcc cggcacgagg aggagtttga tctgttcatg cgcatggacc
     2401 tggaccgcag gcgcgaggag gcccgcaacc ccaagcggaa gccgcgcctc atggaggagg
     2461 acgagctccc ctcgtggatc atcaaggacg acgcggaggt ggagcggctg acctgtgagg
     2521 aggaggagga gaagatgttc ggccgtggct cccgccaccg caaggaggtg gactacagcg
     2581 actcactgac ggagaagcag tggctcaaga aaattacagg aaaagatatc catgacacag
     2641 ccagcagtgt ggcacgtggg ctacaattcc agcgtggcct tcagttctgc acacgtgcgt
     2701 caaaggccat cgaggagggc acgctggagg agatcgaaga ggaggtccgg cagaagaaat
     2761 catcacggaa gcgcaagcga gacagcgacg ccggctcctc caccccgacc accagcaccc
     2821 gcagccgcga caaggacgac gagagcaaga agcagaagaa gcgcgggcgg ccgcctgccg
     2881 agaaactctc ccctaaccca cccaacctca ccaagaagat gaagaagatt gtggatgccg
     2941 tgatcaagta caaggacagc agcagtggac gtcagctcag cgaggtcttc atccagctgc
     3001 cctcgcgaaa ggagctgccc gagtactacg agctcatccg caagcccgtg gacttcaaga
     3061 agataaagga gcgcattcgc aaccacaagt accgcagcct caacgaccta gagaaggacg
     3121 tcatgctcct gtgccagaac gcacagacct tcaacctgga gggctccctg atctatgaag
     3181 actccatcgt cttgcagtcg gtcttcacca gcgtgcggca gaaaatcgag aaggaggatg
     3241 acagtgaagg cgaggagagt gaggaggagg aagagggcga ggaggaaggc tccgaatccg
     3301 aatctcggtc cgtcaaagtg aagatcaagc ttggccggaa ggagaaggca caggaccggc
     3361 tgaagggcgg ccggcggcgg ccgagccgag ggtcccgagc caagccggtc gtgagtgacg
     3421 atgacagtga ggaggaacaa gaggaggacc gctcaggaag tggcagcgaa gaagactgta
     3481 cagactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     3541 atgacaagta actcgagata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3601 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3661 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttagt
     3721 tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3781 gttgggcact gacaattccg tggtgtttat ttgtgaaatt tgtgatgcta ttgctttatt
     3841 tgtaaccatc tagctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3901 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     3961 gggagatgtg ggaggttttt taaagatctg gagaccggtg tggcacgtgc ggaccgagcg
     4021 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4081 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4141 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     4201 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     4261 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     4321 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4381 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4441 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4501 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4561 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     4621 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4681 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4741 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4801 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4861 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4921 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4981 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5041 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5101 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5161 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5221 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5281 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5341 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5401 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5461 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5521 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5581 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5641 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5701 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5761 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5821 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5881 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5941 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6001 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6061 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6121 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6181 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6241 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6301 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6361 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6421 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6481 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6541 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6601 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6661 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6721 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
