LOCUS       sequence-479507-        7057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for KDM5B expression .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7057)
  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  JOURNAL   Unpublished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 7057)
  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            "Unpublished"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-EF1alpha-5' KDM5B
FEATURES             Location/Qualifiers
     source          1..7057
                     /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"
     CDS             1076..1087
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     repeat_region   4320..4460
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4320..4449
                     /label=AAV2 ITR
     rep_origin      4535..4990
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4622..4641)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4832..4853
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5007..5026)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5126..5148
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5186..5204)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5272..5376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5377..5445,5446..6237)
                     /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(5595..5614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6408..6996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6897..6916
                     /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 ctggatctga ggcggccacc acactgcacc caggcccgcg
      781 cccggcgctg cccctcgggg gcccgggccc gctgggcgag ttcctgcctc cacccgagtg
      841 cccggtcttc gaacccagct gggaagagtt cgcggacccc ttcgctttca tccacaagat
      901 ccggcccata gccgagcaga ctggcatctg taaggtgcgg ccgccgccgg attggcagcc
      961 accatttgca tgtgatgttg ataaacttca ttttacgcca cgtatccaga gactgaatga
     1021 attggaggcc caaactcgtg taaaattgaa tttcttggac cagattgcaa agtactggga
     1081 gttacaggga agtactctga aaattccaca tgtggagagg aagatcttgg acttatttca
     1141 gcttaataag ttagttgcag aagaaggtgg atttgcagtt gtttgcaagg atagaaaatg
     1201 gaccaaaatt gctaccaaga tggggtttgc tcctggcaaa gcagtgggct cacatatcag
     1261 agggcattat gaacgaattc tcaaccccta caacttattc ctgtccggag acagcctaag
     1321 gtgtttgcag aagccaaacc tgaccacaga cactaaggac aaggagtaca aaccccatga
     1381 tattccccag aggcagtctg tgcagccttc ggaaacgtgc cccccagccc gacgagcaaa
     1441 acgcatgaga gcagaggcca tgaatattaa aatagaaccc gaggagacaa cagaagccag
     1501 aactcataat ctgagacgtc gaatgggttg tccaactcca aaatgtgaaa atgagaaaga
     1561 aatgaagagt agcatcaagc aagaacctat tgagaggaaa gattatattg tagaaaatga
     1621 gaaggaaaag cccaagagtc gatctaaaaa agccaccaat gctgtggacc tgtatgtctg
     1681 tcttttatgt ggcagtggca atgatgaaga ccggctactg ttgtgtgatg gctgtgatga
     1741 cagttaccat accttttgct tgatcccacc tctccatgat gttcccaagg gagactggag
     1801 gtgtcctaag tgtttggctc aggaatgtag taagccacaa gaagcatttg gctttgaaca
     1861 agcagccagg gactataccc tccgtacttt tggggaaatg gcagatgcgt tcaaatctga
     1921 ttacttcaac atgccagtcc atatggtccc cacagagctt gttgagaaag aattttggag
     1981 actagtaagc actattgagg aggatgtcac agtggaatat ggagctgaca ttgcctcaaa
     2041 ggaatttggc agtggctttc ctgtccgaga tgggaaaatc aaactctcac ctgaggaaga
     2101 ggagtatctt gatagtggct ggaatttgaa caacatgcca gtgatggagc agtctgtcct
     2161 tgcacatatt actgctgata tatgtggcat gaaacttcct tggttgtatg tgggaatgtg
     2221 cttttcttca ttctgttggc acattgaaga ccactggagc tattcaatta actacttgca
     2281 ctggggtgag ccaaaaacct ggtatggagt cccagggtat gctgctgagc agctagaaaa
     2341 tgtaatgaag aaactagctc cagaactctt tgtgtcccag ccggatctcc tccatcagct
     2401 tgtgaccatc atgaacccca ataccctgat gactcatgaa gtgcctgttt accgaactaa
     2461 tcagtgtgct ggggagtttg tgattacatt tccaagagcc taccacagtg gttttaacca
     2521 gggttttaat tttgctgagg ctgttaactt ctgcactgtt gattggctgc cattaggccg
     2581 acagtgtgtg gagcattatc gcttgcttca tcgatattgt gtgttttccc atgatgagat
     2641 gatctgcaag atggcttcca aggctgatgt attagatgtt gtagtggctt caactgttca
     2701 gaaagacatg gccattatga ttgaggatga gaaagcttta agagaaactg tccgtaaatt
     2761 gggagtgatt gattcggaaa gaatggattt tgagctgttg ccagatgatg aacgtcagtg
     2821 tgtaaaatgc aaaactacat gcttcatgtc tgccatctcc tgttcttgta aacctggcct
     2881 tcttgtttgc ctgcatcatg taaaagaatt gtgttcctgt cctccttata aatataaatt
     2941 gcggtatagg tacacgctgg atgatctcta ccctatgatg aatgcattga agcttcgagc
     3001 agaatcttac aacgaatggg ccttgaatgt gaatgaagct ttggaggcaa agatcaacaa
     3061 gaagaaaagc cttgtcagct tcaaggcttt aattgaagaa tctgaaatga agaaattccc
     3121 agacaatgat cttttgcgac accttcgcct agtcacacag gatgcagaga agtgtgcctc
     3181 tgttgcgcag cagttgctta atggcaaaag gcaaactaga tatcgatctg gtggagggaa
     3241 atcccaaaat cagttgacag tgaatgagct ccggcagttt gtaacacagc tgtatgctct
     3301 tccatgtgtc ctcagtcaga caccattact aaaggatctc ttgaatcgtg tagaagattt
     3361 tcaacagcat agtcagaaac tactctctga ggaaacgcct agtgctgcgg agctgcagga
     3421 cttgctagat gtcagctttg aatttgatgt tgaacttcca cagcttgctg agatgcgtat
     3481 ccgtttggaa caagcccgtt ggctagaaga ggtgcagcaa gcttgcctag accccagctc
     3541 ccttacttta gatgatatga gacgtctcat agacctaggg gtagggctgg ccccgtattc
     3601 agcagtggag aaagctatgg cccggctgca ggaactgctc acagtgtcag agcactggga
     3661 cgacaaagcc aagagtctcc tcaaggccag gccacgacat tcattgaata gccttgctac
     3721 ggcagtaaag gaaatcgaag agatccctgc atatctgccc aatggtgcgg ctctgaaaga
     3781 ctcagtgcag agagccagag actggcttca ggatgtagag ggcctgcagg ctggaggacg
     3841 tgtgccagtg ttagacacac tcatagaact tgttacacga ggccgatcta tcccggtaca
     3901 tctgaattct ttgccaagac tggaaaccct agtagctgag gttcaggctt ggaaagaatg
     3961 tgctgttaat acattcttga ctgagaattc tccatattct ctcttagagg tgctgtgtcc
     4021 tcgatgtgat attggccttt tgggattgaa aaggaagcag agaaagttaa aggagccctt
     4081 gccaaatgga aagaaaaaaa gcaccaaatt agagagtctg agtgacctgg agagagcttt
     4141 aactgaaagc aaggagactg cttcagctat ggcaactctt ggggaagctc gcctaaggta
     4201 agtatcaagg ttacaagaca ggtttaagga gaccaataag ctattattcg tataggatac
     4261 tttatacgaa gttattaagt tatggagacc ggtgtggcac gtgcggaccg agcggccgca
     4321 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4381 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4441 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4501 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     4561 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     4621 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4681 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4741 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4801 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4861 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     4921 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4981 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5041 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5101 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5161 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5221 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5281 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5341 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5401 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5461 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5521 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5581 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5641 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5701 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5761 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5821 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5881 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5941 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6001 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6061 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6121 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6181 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6241 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6301 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6361 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6421 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6481 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6541 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6601 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6661 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6721 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6781 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6841 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6901 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6961 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7021 cggttcctgg ccttttgctg gccttttgct cacatgt
//
