LOCUS       sequence-489076-        6647 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for SMARCA4 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6647)
  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  JOURNAL   Unpublished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 6647)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAVLINK-EF1alpha-HA-5'-SMARCA4
FEATURES             Location/Qualifiers
     source          1..6647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2594..2620)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             2657..2925
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(2938..3149)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   3206..3335
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3397..3985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3477..3496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4156..4947,4948..5016))
                     /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     4779..4798
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5017..5121)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5189..5207
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5245..5267)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5367..5386
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5403..5858)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5540..5561)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5752..5771
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5933..6073
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5944..6073
                     /label=AAV2 ITR
ORIGIN
        1 tgatgtactc gtacgtcgtc agcaagacgt tgaacttccc actccggagc tgggggacaa
       61 aggcccgtct tgctgctggg gatcccttgt aagacacctt caccacggag ggggcccact
      121 tgtcaaactc gtacgcccag ttggacagcg ttgagagagg cacgatgatg aggaagggcc
      181 cattgatgcg tttgtgctcc atgaggtacg tgatgagcgc gatggtctgg atggtcttcc
      241 ccaggcccat ctcgtcggcc aggatgccgt tcaggttgtt gttgtacagg gacaccagcc
      301 actccaaacc tttgatctgg tactgtttga ggacaccatt gaccataagc gctgactgct
      361 tgtccactct ctcagtgaca gcatgggcca cggcatagta ggactgcagg ccacgtgcaa
      421 gggcctggga cacgccatat tcatcatcga catcttgctt ggcattctca atgatgtgcc
      481 gcgcgtccac ctcagagacg tcatcgctgt ctggatctgg aatcttcttc ttctcctcca
      541 cgggcagggt gggaggctgt gctgcctgcg gctgctcttc ctcctcctcc tcttcctctt
      601 cttctgagcc actttcttca ctatcagacc tcggagctac ttcatacccc gggttcatct
      661 cgagccaggc ctccagctgc ccggctttgg gggcatctgt gcctgtgagg atcttcccac
      721 tctccacgtg gatcaccttc accgggaggt cgctcatctg gctggtctcg tccagaggct
      781 cgccatccgg cccaatggca ggcgtctgtc cttctgcatt ttctgccttc ttctttttct
      841 ttttcttctt tttctccttg gcgacctggg cagccttgtg ctgccgcacc agctccgtga
      901 ggttagccac gtactcgtct gtctgctgca agaggtaggc caggcgcttg tccttcttct
      961 ggtcgatgag cttgcggtac ccctcctcat cttcagccat gagcctccgc atgcgctcct
     1021 tctcgatccg ctcgttctct ttcttctgct cccgctccgt gttggcatgg tacgtggcca
     1081 ctgccttggt cagcttctgg attttgcctg tgacggatct gtgatattcc ttgaaatcct
     1141 tggcatgctg gagaatgcta ttgaggtatt cctggtgctt ctgccggcgc ttgcgctcct
     1201 gctcgatctt ctgctgcttc tccagcttct cagtgatgcg ggcctcgcgc agggactggc
     1261 gcttgctgcg cttgtaggcc ttagcattga gggctgtctc cagcgctgtg tccctccgca
     1321 tgcacaccac cacctcctgg cgcagctgcc tctggaagtt cagcagcctg agggccttga
     1381 gctcaatggt cgctttggtt cgcaaatccc cggccaggga cccgggaagg ttttcaagtt
     1441 cctgaattcg gtgtgcgatg cgagcctgca gcctgtactc gcgctcctgc aggatctcca
     1501 cagggtcgag gccccgcggc ttctggatgg gggtgatgcg gctctgcttc tggtgcagtg
     1561 gcaccatggg cgcgggctgg gccggctgcc cgggggactg ggtctgcggt ggcatcacgg
     1621 gcgaggcggc gggtgggacg gcagggggcg cgggggaagg gcggcccgtt ggctgcgggg
     1681 gaatcagctt ctgaggggtg ctcgtggggg cagcagcatt cgccatgggt ccttcaggcc
     1741 agggcttggg aggccctcca ggaggctggc ctggcatccc ggggggcacg cccgagggtc
     1801 ctgggggagg catgttgggc cctcccatac catgaggcct gctgtaattt ggaggtgccg
     1861 ggccgggacc cgggccgggg ccagggccag ggccagggcc gggtcctgtt gcggacaccg
     1921 agggtggagg tagcgttggc atctgctgct gcatcccggg catcggccgc ttgccctgca
     1981 ccgccatctg caggtggtcg gggaggggct gccccctggc cagcatcttg taggccatga
     2041 tctgagctct gagctggtgc agctggttct ggttaaatgg ggttgggccc cggttctgct
     2101 gccccaaggc ctgggggtca gcaccatcca gcggggcacc tcctggcccg gaagacatct
     2161 ggggccccga agacgggcca ctggctggaa ctggactaga ggcatgctca gagccaccca
     2221 ggggcgaggg gtaaccttgg gagtgctggt ccatggggct gggcgggggc cccatcccag
     2281 catggccccc tgaccgcatc cccattcctt tcatctggtt gtagcgcggg tcgtccgaca
     2341 tgcccttctc atgcatggac tccatgggct tgtgcatctg gtgcatgttg tcctgagggt
     2401 accctccagg cccctgggtg gggatggggt gtcctgctga gggcggccct gggctgggcc
     2461 ccatcatgct gtgggcggag cccggcgagg gacccgggct agggcccagc atggctccag
     2521 gggaagggcc agggcccggg gaaggacctg gccgaggagt tccgcccagg ggtgggtctg
     2581 gagtggaaga tccagcatag tcaggtacgt cataagggta catggtggcc gtacggtagg
     2641 cgccggtcac agcttggatc tgtaacggcg cagaacagaa aacgaaacaa agacgtagag
     2701 ttgagcaagc agggtcaggc aaagcgtgga gagccggctg agtctaggta ggctccaagg
     2761 gagcgccgga caaaggcccg gtctcgacct gagctttaaa cttacctaga cggcggacgc
     2821 agttcaggag gcaccacagg cgggaggcgg cagaacgcga ctcaaccggc gtggatggcg
     2881 gcctcaggta gggcggcggg cgcgtgaagg agagatgcga gcccctcgaa gcttcagctg
     2941 tgttctggcg gcaaacccgt tgcgaaaaag aacgttcacg gcgactactg cacttatata
     3001 cggttctccc ccaccctcgg gaaaaaggcg gagccagtac acgacatcac tttcccagtt
     3061 taccccgcgc caccttctct aggcacccgt tcaattgccg acccctcccc ccaacttctc
     3121 ggggactgtg ggcgatgtgc gctctgccca ctgacgggca ccggagcgat cgcagatcct
     3181 ctagagtcga cacgcgtgcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3241 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc
     3301 ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa
     3361 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3421 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3481 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3541 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3601 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3661 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3721 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3781 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3841 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3901 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3961 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4021 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4081 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4141 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4201 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4261 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4321 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4381 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4441 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4501 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4561 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4621 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4681 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4741 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4801 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4861 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4921 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4981 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5041 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5101 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5161 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     5221 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5281 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5341 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     5401 taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     5461 attttttaac caatagaccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
     5521 gatagagttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     5581 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     5641 caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5701 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5761 agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5821 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct ttgacgtatg
     5881 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg
     5941 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     6001 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     6061 actaggggtt cctgcggccg ctcggtccgc acgtgccaca ccggtctcca taacttaata
     6121 acttcgtata aagtatccta tacgaataat agcttattgg tctccttaaa cctgtcttgt
     6181 aaccttgata cttaccttgg cctgcatgtg gcggtagagc actcgctgca gcgcagacat
     6241 gtcgcacttg atgacgtact ccaccttttc gggcaactgg gcctcgactt ccttcttgag
     6301 tcgtcggagc aagaagggcc gcagcacttt gtggagacgc cggatgatga gaatggtttc
     6361 ctcctcattc aggtccacct tttccccggt catggcaaag ggtgcgttaa accactgctc
     6421 gaaggtgctg cagctcttga agatggtggg cagcaggaag ttgagcagcg cccagagctc
     6481 gggaagcttg ttctgcagcg gtgtgcccgt cagcagcagg cggcggggtg ccacatagtg
     6541 cgtgttgagc acctgcgtca gcttgcagtg gtggttcttc atgcggtgac cttcgtccac
     6601 aatcatgtac ttccaacgga tcttggcgag gatgtgcttg tctttga
//
