LOCUS       sequence-479623-        6821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for KDM5A expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6821)
  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  JOURNAL   Unpublished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 6821)
  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            "Unpublished"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' KDM5A
FEATURES             Location/Qualifiers
     source          1..6821
                     /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             1037..1048
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     repeat_region   4084..4224
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4084..4213
                     /label=AAV2 ITR
     rep_origin      4299..4754
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4386..4405)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4596..4617
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4771..4790)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4890..4912
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4950..4968)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5036..5140
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5141..5209,5210..6001)
                     /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(5359..5378)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6172..6760
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6661..6680
                     /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 ctggatctgc gggcgtgggg ccggggggct acgcggcgga
      781 gttcgtgcca ccgccagagt gccccgtctt tgagccgagt tgggaggagt tcacagatcc
      841 gctcagcttt atcggccgca tccggccttt ggcggagaaa accggcatct gcaaaattcg
      901 gccgcccaag gactggcagc ctccatttgc ctgtgaagta aaaagctttc gtttcactcc
      961 aagagtccag cgcctgaatg aacttgaggc aatgaccaga gtgagattgg atttcttgga
     1021 tcaactagca aaattttggg aacttcaagg atctactctg aagatccctg tggtagagag
     1081 aaaaatcctg gatctgtatg ctttgagcaa gattgttgcc agcaaaggag gttttgaaat
     1141 ggtcaccaaa gagaagaaat ggtctaaagt gggtagtcgc ttgggatatc tgccaggaaa
     1201 aggaactggg tctcttttga agtcacatta tgaaagaatt ctctacccat atgagctttt
     1261 ccagtctggt gtgagcctta tgggtgtgca gatgcctaat ttagatctta aagaaaaagt
     1321 ggagcctgag gttctcagca ctgataccca aacttcccca gagccaggca caaggatgaa
     1381 cattctgccg aagagaacaa gacgtgtgaa gactcagtca gaatctggag atgtgagtag
     1441 aaacacggaa ctgaagaaac ttcagatttt tggggctggg cccaaggttg tgggcttggc
     1501 aatgggaaca aaagataaag aagatgaggt cacccgaaga cgaaaagtta ccaacaggtc
     1561 agacgcattt aacatgcaaa tgagacaacg gaaaggcact ctctctgtta actttgttga
     1621 tctctatgtt tgtatgtttt gtggtcgggg aaacaatgaa gataaattgc ttttgtgtga
     1681 tggatgtgat gacagctatc atacattttg tctaattcct ccactacctg atgtgcccaa
     1741 aggagactgg aggtgtccta aatgtgtcgc cgaggaatgt agcaaacctc gagaagcctt
     1801 tggatttgaa caagctgtac gagagtatac acttcagagc tttggagaga tggcagataa
     1861 ttttaagtct gattatttta atatgccagt ccatatggtt cccacagaac tagtagaaaa
     1921 ggaattttgg cggctggtaa gcagcattga agaagatgtt attgtggaat atggagcaga
     1981 tatctcctca aaagactttg gaagtggatt tccggtgaag gatgggcgga gaaagattct
     2041 gccagaagaa gaggaatatg cactttctgg ttggaatttg aataacatgc ctgtcctgga
     2101 acagtctgtt cttgcacata ttaatgtgga catctctggt atgaaagtgc cgtggctcta
     2161 tgtgggaatg tgcttctctt ctttttgctg gcacattgag gatcactgga gttattccat
     2221 caactacttg cactgggggg agccaaagac atggtatggt gtgccatctc atgctgcaga
     2281 gcaactggag gaggtgatga gagagctggc ccccgagtta tttgaatccc agcctgatct
     2341 tctgcatcag ttagttacca tcatgaaccc caacgtgcta atggagcatg gtgtgcctgt
     2401 gtacaggacc aatcagtgtg ctggcgagtt tgttgtgaca tttcctcgtg cctatcactc
     2461 tggatttaac cagggctaca actttgctga agctgtgaac ttctgtactg ctgactggtt
     2521 gcccattgga cgtcaatgtg taaatcatta ccgacgccta aggcgccact gtgtcttttc
     2581 acacgaggaa ctaattttca agatggcagc agatccagaa tgcttagatg tggggctggc
     2641 tgccatggtc tgcaaagaat tgactctcat gactgaagaa gaaacacgat taagagagtc
     2701 tgttgtacag atgggtgtcc tgatgtcaga agaagaagtg tttgaacttg ttcctgatga
     2761 tgagcggcag tgttcagcat gcagaaccac atgttttctc tctgctctca catgttcctg
     2821 taatcctgag cggcttgtat gtctctacca tccaactgat ctgtgcccct gccccatgca
     2881 gaagaaatgt cttagatatc gctacccatt agaagacctc ccttctctgc tatatggtgt
     2941 aaaagtcagg gcacagtcct atgacacttg ggtcagtcgt gttacagaag cattgtctgc
     3001 taacttcaac cacaaaaaag atttgattga attgcgagta atgctggaag atgctgagga
     3061 taggaaatac ccagagaatg atctctttcg aaaactcagg gatgctgtaa aagaagctga
     3121 gacctgtgct tctgtggctc agctgcttct gagcaaaaag cagaaacaca gacagagccc
     3181 agatagtggg aggactcgga ccaaactgac agtggaagaa ttgaaggcct ttgtccaaca
     3241 actttttagt cttccgtgtg tcatcagcca agctcggcaa gtaaagaatc tgctagatga
     3301 tgtggaagag tttcatgaac gtgctcagga ggccatgatg gatgaaaccc cagattcttc
     3361 caaactccag atgttgatag atatgggctc tagtctctat gtggaactcc ctgaattacc
     3421 acgactgaag caagagctac aacaggctcg gtggttggac gaagtaagac tgaccttatc
     3481 agatccgcaa caagtcactt tggatgtcat gaagaagctg atagactctg gggtagggtt
     3541 ggcaccccac catgctgtgg agaaagcaat ggctgaacta caggagctcc ttacagtctc
     3601 tgaacgatgg gaagaaaagg ctaaggtctg cctacaggca agaccgaggc acagtgtggc
     3661 aagtttagaa agcattgtga atgaagccaa gaacattcca gcctttctac ccaatgtgtt
     3721 gtccttgaaa gaagccttac aaaaggctcg agaatggacc gctaaagtgg aagctattca
     3781 gagtggcagc aattacgctt atttggagca gcttgagagc ttgtctgcga aaggacgccc
     3841 tattcctgtg cgtcttgaag cactgccgca agtggaatca caggtagcag cagcacgggc
     3901 atggagagaa cggactgggc ggacgtttct taagaagaat tctagccata cattgttaca
     3961 ggtaagtatc aaggttacaa gacaggttta aggagaccaa taagctatta ttcgtatagg
     4021 atactttata cgaagttatt aagttatgga gaccggtgtg gcacgtgcgg accgagcggc
     4081 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4141 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4201 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     4261 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4321 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     4381 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4441 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4501 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4561 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4621 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     4681 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4741 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4801 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4861 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4921 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4981 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5041 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5101 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5161 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5221 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5281 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5341 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5401 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5461 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5521 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5581 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5641 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5701 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5761 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5821 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5881 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5941 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6001 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6061 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6121 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6181 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6241 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6301 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6361 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6421 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6481 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6541 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6601 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6661 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6721 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6781 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
