LOCUS       sequence-476891-        6424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for KDM5C expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6424)
  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  JOURNAL   Unpublished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 6424)
  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            "Unpublished"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' KDM5C
FEATURES             Location/Qualifiers
     source          1..6424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(474..500)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             537..805
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(818..1029)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   1074..1203
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1265..1853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1345..1364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2024..2815,2816..2884))
                     /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     2647..2666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2885..2989)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3057..3075
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3113..3135)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3235..3254
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3271..3726)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3408..3429)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3620..3639
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3801..3941
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3812..3941
                     /label=AAV2 ITR
     protein_bind    5884..5895
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
ORIGIN
        1 tagtgggagc gtagcaagga gccaatattt ttgcctggtg gatagttgag gcgctgggct
       61 acccgagccc accgacggtc cttgcagata gcttcataac caccttcctc caccacaatt
      121 ttgctgagac tgtagaggtc caagatccgc cgttctacat tgggaatctt taaggaggag
      181 ccctggattt cccagaattt ggcaatctgg tccaagtagt tcagtttcac tctcgtctgg
      241 gcctctagct cattcagcct ctggattcgg ggggtaaacc tgaagttgtc cacttccaca
      301 gcaaagggtg gctgccagtc cgcgggtggg cggatcttgc aaatgcccga tttctctgcg
      361 atgggcctga ttttcgcgat gtagccaaga gggtctcgga actcggccca gctaggctcg
      421 aacaccgggc actccggtgg cggtaggaaa tcgtcggacc ccggctcaga tccagcatag
      481 tcaggtacgt cataagggta catggtggcc gtacggtagg cgccggtcac agcttggatc
      541 tgtaacggcg cagaacagaa aacgaaacaa agacgtagag ttgagcaagc agggtcaggc
      601 aaagcgtgga gagccggctg agtctaggta ggctccaagg gagcgccgga caaaggcccg
      661 gtctcgacct gagctttaaa cttacctaga cggcggacgc agttcaggag gcaccacagg
      721 cgggaggcgg cagaacgcga ctcaaccggc gtggatggcg gcctcaggta gggcggcggg
      781 cgcgtgaagg agagatgcga gcccctcgaa gcttcagctg tgttctggcg gcaaacccgt
      841 tgcgaaaaag aacgttcacg gcgactactg cacttatata cggttctccc ccaccctcgg
      901 gaaaaaggcg gagccagtac acgacatcac tttcccagtt taccccgcgc caccttctct
      961 aggcacccgt tcaattgccg acccctcccc ccaacttctc ggggactgtg ggcgatgtgc
     1021 gctctgccca ctgacgggca ccggagcgat cgcagatcca cgcgtgcggc cgcaggaacc
     1081 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     1141 accaaaggtc gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     1201 aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1261 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1321 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1381 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1441 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1501 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1561 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1621 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1681 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1741 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1801 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1861 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1921 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1981 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2041 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2101 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2161 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2221 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     2281 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     2341 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     2401 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     2461 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     2521 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     2581 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     2641 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     2701 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     2761 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     2821 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     2881 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     2941 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3001 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     3061 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     3121 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     3181 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     3241 cagagcagat tgtactgaga gtgcaccata aaattgtaaa cgttaatatt ttgttaaaat
     3301 tcgcgttaaa tttttgttaa atcagctcat tttttaacca atagaccgaa atcggcaaaa
     3361 tcccttataa atcaaaagaa tagcccgaga tagagttgag tgttgttcca gtttggaaca
     3421 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3481 gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttggggtcg aggtgccgta
     3541 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3601 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag gcgctggcaa
     3661 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3721 gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat gcgtaaggag
     3781 aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     3841 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     3901 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgct cggtccgcac
     3961 gtgccacacc ggtctccata acttaataac ttcgtataaa gtatcctata cgaataatag
     4021 cttattggtc tccttaaacc tgtcttgtaa ccttgatact taccttcggg ctgagggggc
     4081 cagtgtgcgt ttcacctcat ccagccatcg cgcctgttcc acctgccgct ggagctgctg
     4141 ggcctcaggc acctccaccc ccagctgccg ccccctctcc aacagggact gcagtagccc
     4201 tggactggag ggcagtgagg ccagggcctc acgagcctca gcctggtagg cctccacctg
     4261 ttccagaaca cccttgacat ccccaatctg gtgcatggcg caaggcaggt tgttcatctg
     4321 gtccagaaag gcccggagct cagtcagggt catctgtaga ccagccaccc tgtgggggcc
     4381 agcttcctgg ccgctgacca gtcccagagc tcgggacacg caagcctctg cctcactcag
     4441 gcagttcttt agttgctgca gcagctcact attaggaaac ctccgctcac gggcttcaga
     4501 ctctagtgcc ctcagttctt caaggctgcg cttccgccca tcctccacct ccagggccac
     4561 tcgcactttg ttggcccagg tgtcaaagga ctcagcccga accttcagct tatgcagcat
     4621 ggcaggaagc tcatccaagg tataccgata ccgcaggtac tgccggctac tggagcactt
     4681 gcagagatca ttgatgtggg aaaggcagac aaggccgtct gggcagtcgt agcaggccag
     4741 ggctgacagg aaacacgtag tcttgcactt gatacactgg cgctcatcat ctgggagcag
     4801 ctcgaaagcc tctcgctcag cctctgtgat acccttctcc agcagggcct ttcgtagacg
     4861 ccgctcttct tgcaccatga tgaacatctc cttatgcaca gctgccgcca ggttcaggtc
     4921 tagcttctct gggcaggcag ccatcttgca gataagctcc tcatgggaga agacgcagta
     4981 tctccggagc cggcggtagt gctcaatgca ctggcgccca gcaggcaacc agtcagcagt
     5041 gcaaaagttg acagcctcgg caaagttgta gccttggttg aagccgctgt ggtaagcacg
     5101 ggggaaggtg atgacaaact ctcctgcaca ctggtttgtg cggacaactg gcacaccatg
     5161 ggacatgagg gtgttgggat tcatgagggt gacaagttgg tgcaggaggt caggctggct
     5221 atcaaatagt tcaggtgtca gcttcttcat cacttcttcc aaatgttctg ctgcaagtga
     5281 gggcacccca taccaggtct tcggctcacc ccagtggagg tagttaatgg agtaactcca
     5341 gtgatcctca atatgccagc aaaaggctga gaagaccatg cccacgtaga gccagggcac
     5401 cttcatgcca gagatatctg cattgatgtg gcacagtaca gactgttcca acaccggcat
     5461 cacatttagg ttccaaccac tggtagcata ctcctcctct tcgggggtta ggtgccgttt
     5521 actgtcactg acagggaaac cgctgccaaa ttctttggaa tggatgtcag ctccatactc
     5581 aacagtcaca tcttcctcaa tgctatttac cagcctccag aactccttct ccacaagttc
     5641 tgtgggcacc atatgcacgg gcatgttgaa gtagtcagct ttaaaggagt cggccatctc
     5701 gccaaagctc tgcagagtgt attcccgggt agcctgctca aagccaaagg cttctggggg
     5761 ccgcttacac tccgccatga cacactttgg gcaccgccag acacccttgg ggatctcagg
     5821 cagaggaggc agcaggcaga agatgtggta gttgtcatca cagccatcac acagcaggag
     5881 cttgtcatcc tcatcccctc gagaacacat ccggcagaca tatgactcaa taaactgggc
     5941 attgctgtgg ttcctccgta gcctcatggt catcttggtg cagggttctg ggctgtgact
     6001 cagctcctcc ttgctctcca ggaaggtctt aggcgatgtt gactccacct tcacatcccc
     6061 acctaactcc tccttcacca ctactgtggg gggacactca ggcccctcct tatctttctt
     6121 ccgcagagtc ttgtctttgg ccatgaggcc caggcccatc atcttggggc ctgccccata
     6181 gatctgtagc tttttcagct ctggattctt ctcaatgtct tcctctgtgg gttccggatc
     6241 aggctgcagt ctcttggccc gccggccata gctgttgaac ttggaaggct gcacagactg
     6301 tcgtaggggg atgctgtggg gtttgtattc cttgtccttc tcctcattat caaatggacg
     6361 tgtgttacac tgcacaaggt tggctccaga ctggtacatt tcatagggat aaacaatgcg
     6421 ttcg
//
