LOCUS       sequence-479622-        5702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for KDM6A expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5702)
  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  JOURNAL   Unpublished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 5702)
  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            "Unpublished"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-5' KDM6A
FEATURES             Location/Qualifiers
     source          1..5702
                     /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        187..398
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             411..679
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             716..742
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     repeat_region   2965..3105
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2965..3094
                     /label=AAV2 ITR
     rep_origin      3180..3635
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3267..3286)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3477..3498
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3652..3671)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3771..3793
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3831..3849)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3917..4021
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4022..4090,4091..4882)
                     /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(4240..4259)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5053..5641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5542..5561
                     /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 gcgtgtcgac tctagaggat ctgcgatcgc tccggtgccc
      181 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      241 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      301 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      361 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
      421 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
      481 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
      541 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
      601 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
      661 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac cgtacggcca ccatgtaccc
      721 ttatgacgta cctgactatg ctggatctaa atcctgcgga gtgtcgctcg ctaccgccgc
      781 cgctgccgcc gccgctttcg gtgatgagga aaagaaaatg gcggcgggaa aagcgagcgg
      841 cgagagcgag gaggcgtccc ccagcctgac agccgaggag agggaggcgc tcggcggact
      901 ggacagccgc ctctttgggt tcgtgagatt tcatgaagat ggcgccagga cgaaggccct
      961 actgggcaag gctgttcgct gctatgaatc tctaatctta aaagctgaag gaaaagtgga
     1021 gtctgatttc ttttgtcaat taggtcactt caacctctta ttggaagatt atccaaaagc
     1081 attatctgca taccagaggt actacagttt acagtctgac tactggaaga atgctgcctt
     1141 tttatatggt cttggtttgg tctacttcca ttataatgca tttcagtggg caattaaagc
     1201 atttcaggag gtgctttatg ttgatcccag cttttgtcga gccaaggaaa ttcatttacg
     1261 acttgggctt atgttcaaag tgaacacaga ctatgagtct agtttaaagc attttcagtt
     1321 agctttggtt gactgtaatc cctgcacttt gtccaatgct gaaattcaat ttcacattgc
     1381 ccacttatat gaaacccaga ggaaatatca ttctgcaaaa gaagcttatg aacaactttt
     1441 gcagacagag aatctttctg cacaagtaaa agcaactgtc ttacaacagt taggttggat
     1501 gcatcacact gtagatctcc tgggagataa agccaccaag gaaagctatg ctattcagta
     1561 tctccaaaag tccttggaag cagatcctaa ttctggccag tcctggtatt tcctcggaag
     1621 gtgctattca agtattggga aagttcagga tgcctttata tcttacaggc agtctattga
     1681 taaatcagaa gcaagtgcag atacatggtg ttcaataggt gtgctatatc agcagcaaaa
     1741 tcagcccatg gatgctttac aggcctatat ttgtgctgta caattggacc atggccatgc
     1801 tgcagcctgg atggacctag gcactctcta tgaatcctgc aaccagcctc aggatgccat
     1861 taaatgctac ttaaatgcaa ctagaagcaa aagttgtagt aatacctctg cacttgcagc
     1921 acgaattaag tatttacagg catgtaaacc tcatcatcca aatactgaac ctgtattagg
     1981 cctcagtcaa acaccaattt cacagcaatc cttgccacta cacatgattc cttctagcca
     2041 agtagatgac ctgtccagtc ctgccaagag gaaaagaaca tctagtccaa caaagaatac
     2101 ttctgacaat tggagtggtg gacatgctgt gtcacatcct ccagtacagc aacaagctca
     2161 ttcatggtgt ttgacaccac agaaattaca gcatttggaa cagctccgcg caaatagaaa
     2221 taatttaaat ccagcacaga aactgatgct ggaacagctg gaaagtcagt ttgtcttaat
     2281 gcaacaacac caaatgagac caacaggagt tgcacaggta cgatctactg gaattcctaa
     2341 tgggccaaca gctgactcat cactgcctac aaactcagtc tctggccagc agccacagct
     2401 tgctctgacc agagtgccta gcgtctctca gcctggagtc cgtcctgcct gccctgggca
     2461 gcctttggcc aatggaccct tttctgcagg ccatgttccc tgtagcacat caagaacgct
     2521 gggaagtaca gacactattt tgataggcaa taatcatata acaggaagtg gaagtaatgg
     2581 aaacgtgcct tacctgcagc gaaacgcact cactctacct cataaccgca caaacctgac
     2641 cagcagcgca gaggagccgt ggaaaaacca actatctaac tccactcagg ggcttcacaa
     2701 aggtcagagt tcacattcgg caggtcctaa tggtgaacga cctctctctt ccactgggcc
     2761 ttcccagcat ctccaggcag ctggctctgg tattcagaat cagaacggac atcccaccct
     2821 gcctagcaat tcagtaacac aggtaagtat caaggttaca agacaggttt aaggagacca
     2881 ataagctatt attcgtatag gatactttat acgaagttat taagttatgg agaccggtgt
     2941 ggcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     3001 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3061 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     3121 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     3181 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3241 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3301 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3361 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     3421 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3481 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     3541 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3601 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     3661 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     3721 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3781 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     3841 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     3901 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3961 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4021 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4081 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4141 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4201 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4261 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4321 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4381 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     4441 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     4501 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4561 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     4621 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4681 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4741 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4801 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     4861 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     4921 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4981 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5041 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5101 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5161 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5221 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5281 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5341 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5401 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5461 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5521 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5581 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5641 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5701 gt
//
