LOCUS       sequence-491932-        6422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for CHD1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6422)
  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  JOURNAL   Unpublished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 6422)
  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            "Unpublished"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: pAAVLINK-EF1alpha-HA-5'-CHD1
FEATURES             Location/Qualifiers
     source          1..6422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..386
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             399..667
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             704..730
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      3900..4355
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3987..4006)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4197..4218
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4372..4391)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4491..4513
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4551..4569)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4637..4741
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4742..4810,4811..5602)
                     /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(4960..4979)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5773..6361
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6262..6281
                     /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 gcgtggatct gcgatcgctc cggtgcccgt cagtgggcag
      181 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
      241 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      301 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      361 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
      421 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
      481 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
      541 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
      601 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
      661 acagatccaa gctgtgaccg gcgcctaccg tacggccacc atgtaccctt atgacgtacc
      721 tgactatgct ggatctaatg gacacagtga tgaagaaagt gttagaaaca gtagtggaga
      781 atcaagccag tcggatgatg attctgggtc agcttcaggc tctggatctg gttcgagttc
      841 tggaagcagt agtgatggaa gcagtagcca gtcaggtagc agtgactctg actccggatc
      901 tgaatcaggc agtcagtcag agtctgagtc agacacttcc cgagaaaaca aagttcaagc
      961 aaaaccaccg aaagttgatg gagctgagtt ttggaaatct agtcctagta ttctggccgt
     1021 tcagagatct gcaatcctca agaagcagca acagcagcag cagcaacaac aacatcaagc
     1081 ctcatctaat agcggatcag aagaggattc ctctagcagt gaagattccg atgactcatc
     1141 aagtgaggtc aaaaggaaaa agcataaaga tgaagattgg caaatgtctg ggtcaggatc
     1201 tccatctcag tctggttcag attcagaatc tgaagaagag agagagaaaa gcagttgtga
     1261 tgaaacagaa tctgattatg agccaaaaaa caaagtcaaa agcagaaaac ctcaaaatag
     1321 atctaagtca aaaaatggaa agaagattct tggacaaaaa aagagacaga ttgattcatc
     1381 tgaggaggat gatgatgaag aagattatga taatgataaa agaagttctc gtcgccaagc
     1441 aactgttaat gttagctata aggaggatga agaaatgaaa acagattctg atgacctact
     1501 ggaagtctgt ggagaggatg ttcctcaacc tgaggaagag gaatttgaaa ccatagaaag
     1561 atttatggat tgtcggattg ggagaaaagg agctactggt gctactacaa ccatctatgc
     1621 agttgaagca gatggtgacc caaatgcagg ctttgaaaaa aacaaagaac caggagagat
     1681 tcagtattta attaaatgga aaggatggtc ccatatccac aacacttggg agacagaaga
     1741 aaccctcaag cagcagaatg ttagaggaat gaaaaaattg gataattata agaaaaaaga
     1801 tcaggaaaca aaaagatggt tgaaaaatgc ctctccagaa gatgtggaat attataattg
     1861 ccagcaagaa cttacagatg atctacataa acagtatcaa atagtggaac gtataattgc
     1921 tcattccaat caaaagtcag cagctggtta tcctgattat tactgcaaat ggcagggcct
     1981 tccatactca gagtgcagct gggaagatgg agctctcatt tccaaaaagt ttcaagcatg
     2041 cattgatgag tattttagca ggaaccaatc aaaaaccact ccttttaaag attgcaaagt
     2101 attaaaacaa aggccaaggt ttgtagccct gaagaagcag ccatcctata ttggaggaca
     2161 tgagggctta gaattaagag attatcaact gaatggttta aattggcttg ctcattcttg
     2221 gtgcaaagga aatagttgca tactcgctga tgaaatgggc cttggaaaaa caatacagac
     2281 gatctcattt ctgaattatt tgtttcatga acatcaatta tatggacctt ttttattggt
     2341 agtaccgctc tccactctta cttcctggca aagggaaatt cagacttggg cttctcaaat
     2401 gaatgctgtg gtttatttag gtgacattaa cagcagaaac atgataagaa ctcatgaatg
     2461 gacgcatcat cagaccaaac ggttaaaatt taatatattg ttaacaactt atgaaatttt
     2521 attaaaagat aaggcattcc ttggaggtct aaattgggca tttataggtg ttgatgaagc
     2581 acaccgatta aagaatgatg actcccttct gtataaaact ttaatagatt ttaaatccaa
     2641 tcatcgtctc cttatcactg gaactcctct acagaattcc ctcaaagagc tctggtcttt
     2701 gctacatttc attatgccag aaaagttttc ttcctgggaa gattttgaag aagaacatgg
     2761 caaagggaga gaatatggtt atgcaagcct tcacaaggag cttgagccat ttctgttacg
     2821 ccgagttaag aaagatgtgg aaaaatctct tcctgccaag gttgagcaga ttttaagaat
     2881 ggaaatgagt gctttacaga aacaatatta caaatggatt ttaactagga attacaaagc
     2941 cctcagcaaa ggttccaagg gcagtacctc aggctttttg aacattatga tggagctaaa
     3001 gaaatgttgt aaccattgct acctcattaa accaccagat aataatgaat tctataataa
     3061 acaggaggcc ttacaacact taattcgtag tagcggaaaa ttgattcttc ttgacaagct
     3121 attaattcgc ctaagagaac gaggcaatcg agttcttatt ttttcacaaa tggtgcggat
     3181 gttagatata cttgcagaat atttgaaata tcgtcaattc ccctttcaaa gattagatgg
     3241 atcaataaaa ggagaactga ggaaacaagc tctagatcat tttaatgctg agggatcaga
     3301 ggatttttgc tttttgctgt ccacaagagc tggaggtcta gggattaatt tagcctctgc
     3361 tgacactgtt gttatatttg attccgattg gaatccacag aatgatcttc aggcacaggc
     3421 tagagcccat cgaattgggc aaaagaaaca ggtgaatatt tatcgtctag ttacaaaggg
     3481 atcagttgaa gaagatattc ttgaaagggc gaaaaagaag atggttttag atcatcttgt
     3541 aattcaaaga atggacacaa ctgggaagac agtactacat acaggtaagt atcaaggtta
     3601 caagacaggt ttaaggagac caataagcta ttattcgtat aggatacttt atacgaagtt
     3661 attaagttat ggagaccggt gtggcacgtg cggaccgagc ggccgcagga acccctagtg
     3721 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     3781 gtcgcccggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     3841 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     3901 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3961 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4021 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4081 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4141 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4201 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     4261 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4321 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     4381 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4441 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4501 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4561 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4621 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4681 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4741 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4801 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4861 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4921 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4981 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5041 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5101 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5161 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5221 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5281 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5341 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5401 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5461 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5521 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5581 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5641 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5701 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5761 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5821 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5881 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5941 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6001 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6061 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6121 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6181 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6241 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6301 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6361 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6421 gt
//
