LOCUS       sequence-491405-        6582 bp    DNA     circular SYN 03-NOV-2025
DEFINITION  5' AAVLINK plasmid for KDM6B expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6582)
  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  JOURNAL   Unpublished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 6582)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 3, 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            "Unpublished"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'-KDM6B
FEATURES             Location/Qualifiers
     source          1..6582
                     /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"
     CDS             complement(1128..1139)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1488..1517
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1491..1520
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1494..1523
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1497..1526
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1500..1529
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2992..3021
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2995..3024
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     rep_origin      4060..4515
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4147..4166)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4357..4378
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4532..4551)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4651..4673
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4711..4729)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4797..4901
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4902..4970,4971..5762)
                     /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(5120..5139)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5933..6521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6422..6441
                     /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 ggatctcatc gggcagtgga ccctccaggg gcccgcgctg cacgggaagc
      781 ctttgccctt gggggcctga gctgtgctgg ggcctggagc tcctgcccgc ctcatccccc
      841 tcctcgtagc gcatggctgc ctggaggcag atgctcagcc agcattgggc agcccccgct
      901 tcctgctccc ctaccccctt cacatggcag tagttctggg caccccagca aaccatatta
      961 tgctccaggg gcgcccactc caagacccct ccatgggaag ctggaatccc tgcatggctg
     1021 tgtgcaggca ttgctccggg agccagccca gccagggctt tgggaacagc ttgggcaact
     1081 gtacgagtca gagcacgata gtgaggaggc cacacgctgc taccacagcg cccttcgata
     1141 cggaggaagc ttcgctgagc tggggccccg cattggccga ctgcagcagg cccagctctg
     1201 gaactttcat actggctcct gccagcaccg agccaaggtc ctgcccccac tggagcaagt
     1261 gtggaacttg ctacaccttg agcacaaacg gaactatgga gccaagcggg gaggtccccc
     1321 ggtgaagcga gctgctgaac ccccagtggt gcagcctgtg cctcctgcag cactctcagg
     1381 cccctcaggg gaggagggcc tcagccctgg aggcaagcga aggagaggct gcaactctga
     1441 acagactggc cttcccccag ggctgccact gcctccacca ccattaccac caccaccacc
     1501 accaccacca ccaccaccac caccaccacc cctgcctggc ctggctacca gccccccatt
     1561 tcagctaacc aagccagggc tgtggagtac cctgcatgga gatgcctggg gcccagagcg
     1621 caagggttca gcacccccag agcgccagga gcagcggcac tcgctgcctc acccatatcc
     1681 atacccagct ccagcgtaca ccgcgcaccc ccctggccac cggctggtcc cggctgctcc
     1741 cccaggccca ggcccccgcc ccccaggagc agagagccat ggctgcctgc ctgccacccg
     1801 tccccccgga agtgacctta gagagagcag agttcagagg tcgcggatgg actccagcgt
     1861 ttcaccagca gcaaccaccg cctgcgtgcc ttacgcccct tcccgccccc ctggcctccc
     1921 cggcaccacc accagcagca gcagtagcag cagcagcaac actggtctcc ggggcgtgga
     1981 gccgaaccca ggcattcccg gcgctgacca ttaccaaact cccgcgctgg aggtctctca
     2041 ccatggccgc ctggggccct cggcacacag cagtcggaaa ccgttcttgg gggctcccgc
     2101 tgccactccc cacctatccc tgccacctgg accttcctca ccccctccac ccccctgtcc
     2161 ccgcctctta cgccccccac caccccctgc ctggttgaag ggtccggcct gccgggcagc
     2221 ccgagaggat ggagagatct tagaagagct cttctttggg actgagggac ccccccgccc
     2281 tgccccacca cccctccccc atcgcgaggg cttcttgggg cctccggcct cccgcttttc
     2341 tgtgggcact caggattctc acacccctcc cactccccca accccaacca ccagcagtag
     2401 caacagcaac agtggcagcc acagcagcag ccctgctggg cctgtgtcct ttcccccacc
     2461 accctatctg gccagaagta tagaccccct tccccggcct cccagcccag cacagaaccc
     2521 ccaggaccca cctcttgtac ccctgactct tgccctgcct ccagcccctc cttcctcctg
     2581 ccaccaaaat acctcaggaa gcttcaggcg cccggagagc ccccggccca gggtctcctt
     2641 cccaaagacc cccgaggtgg ggccggggcc acccccaggc cccctgagta aagcccccca
     2701 gcctgtgccg cccggggttg gggagctgcc tgcccgaggc cctcgactct ttgattttcc
     2761 ccccactccg ctggaggacc agtttgagga gccagccgaa ttcaagatcc tacctgatgg
     2821 gctggccaac atcatgaaga tgctggacga atccattcgc aaggaagagg aacagcaaca
     2881 acacgaagca ggcgtggccc cccaaccccc gctgaaggag ccctttgcat ctctgcagtc
     2941 tcctttcccc accgacacag cccccaccac tactgctcct gctgtcgccg tcaccaccac
     3001 caccaccacc accaccacca ccacggccac ccaggaagag gagaagaagc caccaccagc
     3061 cctaccacca ccaccgcctc tagccaagtt ccctccaccc tctcagccac agccaccacc
     3121 acccccaccc cccagcccgg ccagcctgct caaatccttg gcctccgtgc tggagggaca
     3181 aaagtactgt tatcggggga ctggagcagc tgtttccacc cggcctgggc ccttgcccac
     3241 cactcagtat tcccctggcc ccccatcagg tgctaccgcc ctgccgccca cctcagcggc
     3301 ccctagcgcc cagggctccc cacagccctc tgcttcctcg tcatctcagt tctctacctc
     3361 aggcgggccc tgggcccggg agcgcagggc gggcgaagag ccagtcccgg gccccatgac
     3421 ccccacccaa ccgcccccac ccctatctct gccccctgct cgctctgagt ctgaggtgct
     3481 agaagagatc agccgggctt gcgagaccct tgtggagcgg gtgggccgga gtgccactga
     3541 cccagccgac ccagtggaca cagcagagcc agcggacagt gggactgagc gactgctgcc
     3601 ccccgcacag gccaaggagg aggctggcgg ggtggcggca gtgtcaggca gctgtaagcg
     3661 gcgacagaag gagcatcaga aggagcatcg gcggcacagg cgggcctgta aggacagtgt
     3721 gggtcgtcgg ccccgtgagg gcaggtaagt atcaaggtta caagacaggt ttaaggagac
     3781 caataagcta ttattcgtat aggatacttt atacgaagtt attaagttat ggagaccggt
     3841 gtggcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     3901 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccggc ctcagtgagc
     3961 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     4021 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4081 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     4141 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4201 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4261 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4321 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4381 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4441 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4501 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4561 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4621 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4681 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4741 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4801 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4861 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4921 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4981 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5041 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5101 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5161 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5221 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5281 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5341 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5401 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5461 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5521 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5581 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5641 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5701 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5761 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5821 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5881 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5941 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6001 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6061 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6121 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6181 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6241 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6301 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6361 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6421 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6481 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6541 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
