LOCUS       sequence-476879-        6629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' AAVLINK plasmid for KDM5C expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6629)
  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  JOURNAL   Unpublished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 6629)
  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            "Unpublished"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: pAAVLINKv1-pCALM1-Cre-3' KDM5C
FEATURES             Location/Qualifiers
     source          1..6629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(417..1208,1209..1277))
                     /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     1040..1059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1278..1382)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1450..1468
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1506..1528)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1628..1647
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1664..2119)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1801..1822)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2013..2032
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2194..2334
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2205..2334
                     /label=AAV2 ITR
     primer_bind     2727..2747
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2824..2889)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(4918..5949)
                     /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"
     repeat_region   6096..6225
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(join(6287..6629,1..246))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6367..6386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
       61 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
      121 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
      181 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
      241 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
      301 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
      361 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
      421 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
      481 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
      541 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
      601 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
      661 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
      721 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
      781 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
      841 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
      901 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
      961 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     1021 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     1081 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     1141 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     1201 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     1261 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     1321 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     1381 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     1441 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     1501 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     1561 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     1621 aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat
     1681 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc
     1741 gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt
     1801 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     1861 accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg
     1921 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     1981 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2041 aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2101 gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca
     2161 gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc
     2221 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     2281 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
     2341 gctcggtccg cacgtgccac accggtctcc agatctttaa aaaacctccc acatctcccc
     2401 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2461 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagctag atggttacaa
     2521 ataaagcaat agcatcacaa atttcacaaa taaacaccac ggaattgtca gtgcccaaca
     2581 gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaagaa
     2641 ctaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
     2701 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
     2761 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt atctcgagtc atcagaattc
     2821 ttacttgtca tcgtcatcct tgtagtcgat gtcatgatct ttataatcac cgtcatggtc
     2881 tttgtagtct gtacacaact gttgctgagg cggctgctgt gggcagggca gatgcagccg
     2941 gggagtcaga gtggagaaag gggccgaggg gcctgaagtg gtcccttccg ccggttccaa
     3001 gccattctgg ttctcctggg tgctggggct gccagtggtg gggatgggtg caggggggcc
     3061 ctcaccccca gtctcctcct ccagctcttc ctcctcctgg acctcctcag cctctggccc
     3121 tgagctccgt accctctttg gctctagctc ctctcgggct gggtcatcgc cctccccacc
     3181 ccgatccacc ttccgccgcc gccgcctctc cagggcccgg ccccgagccc gactcccgtg
     3241 acgctctgcc ttctccagct ccagaagtgt gcggatcctc tccaggtctg ggggctgtcc
     3301 agcctgcagc agctgccata tgctgtggtt ctcatccagg gtcacctcga gcaggtcccc
     3361 ctccatcatg agctcctcca agggggcccg ggttgcctca ggcagttcca acacagggcc
     3421 agtcaactgt ggcaacagcg aggacagcag ctccagatct ctcttacctg agccctcctc
     3481 cggggctacc ttctcaggac tggtcacact gtctccattc tccagtaagc cctggacctt
     3541 aggcatatcc ttgccactgc cctctctgag ggggtcagaa gcaggggctg cagggtagtt
     3601 aggaggctcc tcaggtctag gttcagcctg tagccgttgg cggagctcag ccagccgtcc
     3661 caaaagagca gtcacatctt cagaggccag agcctgcctg gcgcggcctt gccagctgat
     3721 ggccctctct gtgaggcact gcagggcctc gccctcgggc agccgcacag gcagtctctg
     3781 cagggctacc agcagtgcca ggatggtctc caggcgcggg cgccttgagc gcatacacag
     3841 tggacacagg aatttggtgt cccattccca ccaggccagc agtggggatg aggtgggatt
     3901 gggcctcgga gagctgagga ggcgaggcac tgacacacac cgcccatgga accagtcctg
     3961 acacaggtca cactgcagag ctcccgcccc agccagcacc tgcccacaca cacagataga
     4021 ggttgtagag gaggccgtgc tcgatgatgc cagtggactg ggcttggccg aattggtgcg
     4081 acgcagctgc aggataccct ccttctcctt ctgttccccc tccttgaagg ccacgatcac
     4141 agagcctggg tccctgaggt cctgcgcaga cagccccagc agctctgtgt cagatttgta
     4201 caaccccagc tccttctcca tccaccggct gcgcttggtg ctgtctgagc cggcatctgc
     4261 acatgggcag agaacctcca gcagcgtgta gcaagaattt ttcttgagga aggtcttgga
     4321 ggccttctcc ctccaggagt gcgctgtcag tacctgtagc tctagctgtc tcagctcctc
     4381 cagccccaca ggtaggtccc ggcccacagc tactaggccc tccaagtcat ccaggcaggg
     4441 gtagtggtca ccattttgga tctcatcaac atcagcaatc caggcccggg ccttagcaag
     4501 agcctccttg agagcctgga tgttgggcag gtgaacaggg atgttttccg cttcacggat
     4561 tatggcctca agtgtggctg gtggatgctt ctgcctggcc tccaggcaga ggtgggcttt
     4621 ctcctcccag cgttcagcaa tggtcagcag ttcctgcagc tcggcctggg ctttatccac
     4681 agcagggcta ggggctacac tggcacccgc gaccaacagt cctcgcatga cagccaaggt
     4741 gcccctgtgg agagaaaggc aaagtggatg tcagtaagac caataggtgc ctatcagaaa
     4801 cgcaagagtc ttctctgtct cgacaagccc agtttcataa attgctataa agtatcctat
     4861 acgaagttat actagttatt ggtctcctta aacctgtctt gtaaccttga tacttaccta
     4921 atcgccatct tccagcaggc gcaccattgc ccctgtttca ctatccaggt tacggatata
     4981 gttcattaca atatttacat tggtccagcc accagcttgc atgatctccg gtattgaaac
     5041 tccagcgcgg gccatatctc gcgcggctcc gacacgggca ctgtgtccag accaggccag
     5101 gtatctctga ccagagtcat ccttagcgcc gtaaatcaat cgatgagttg cttcaaaaat
     5161 cccttccagg gcgcgagttg atagctggct ggtggcagat ggcgcggcaa caccattttt
     5221 tctgacccgg caaaacaggt agttattcgg atcatcagct acaccagaga cggaaatcca
     5281 tcgctcgacc agtttagtta cccccaggct aagtgccttc tctacacctg cggtgctaac
     5341 cagcgttttc gttctgccaa tatggattaa cattctccca ccgtcagtac gtgagatatc
     5401 tttaaccctg atcctggcaa tttcggctat acgtaacagg gtgttataag caatccccag
     5461 aaatgccaga ttacgtatat cctggcagcg atcgctattt tccatgagtg aacgaacctg
     5521 gtcgaaatca gtgcgttcga acgctagagc ctgttttgca cgttcaccgg catcaacgtt
     5581 ttcttttcgg atccgccgca taaccagtga aacagcattg ctgtcacttg gtcgtggcag
     5641 cccggaccga cgatgaagca tgtttagctg gcccaaatgt tgctggatgg tttttactgc
     5701 cagaccgcgc gcctgaagat atagaagata atcgcgaaca tcttcaggtt ctgcgggaaa
     5761 ccatttccgg ttattcaact tgcaccatgc cgcccacgac cggcaaacgg acagaagcat
     5821 tttccaggta tgctcagaaa acgcctggcg atccctgaac atgtccatca ggttcttgcg
     5881 aacctcatca ctcgttgcat cgaccggtaa tgcaggcaaa ttttggtgta cggtcagtaa
     5941 attggacatg gtggcgtacg agtaacggca cggcgtccac gacactccca gcaccactgc
     6001 cggagcgcga gtctgcgccc cgcgatatat atatggtgcg ccgtatccct ccgccgcggc
     6061 ggagccgcct cgcgcagagc gacgcgtgcg gccgcaggaa cccctagtga tggagttggc
     6121 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     6181 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag
     6241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6601 ccaacccggt aagacacgac ttatcgcca
//
