LOCUS       sequence-476868-        6634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' AAVLINK plasmid for KDM5A expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6634)
  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  JOURNAL   Unpublished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 6634)
  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            "Unpublished"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' KDM5A
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..1032)
                     /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   1179..1308
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1370..1958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1450..1469)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2129..2920,2921..2989))
                     /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     2752..2771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2990..3094)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3162..3180
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3218..3240)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3340..3359
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3376..3831)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3513..3534)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3725..3744
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3906..4046
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3917..4046
                     /label=AAV2 ITR
     primer_bind     4439..4459
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4536..4601)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6460..6479)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 ctaatcgcca tcttccagca ggcgcaccat tgcccctgtt tcactatcca ggttacggat
       61 atagttcatt acaatattta cattggtcca gccaccagct tgcatgatct ccggtattga
      121 aactccagcg cgggccatat ctcgcgcggc tccgacacgg gcactgtgtc cagaccaggc
      181 caggtatctc tgaccagagt catccttagc gccgtaaatc aatcgatgag ttgcttcaaa
      241 aatcccttcc agggcgcgag ttgatagctg gctggtggca gatggcgcgg caacaccatt
      301 ttttctgacc cggcaaaaca ggtagttatt cggatcatca gctacaccag agacggaaat
      361 ccatcgctcg accagtttag ttacccccag gctaagtgcc ttctctacac ctgcggtgct
      421 aaccagcgtt ttcgttctgc caatatggat taacattctc ccaccgtcag tacgtgagat
      481 atctttaacc ctgatcctgg caatttcggc tatacgtaac agggtgttat aagcaatccc
      541 cagaaatgcc agattacgta tatcctggca gcgatcgcta ttttccatga gtgaacgaac
      601 ctggtcgaaa tcagtgcgtt cgaacgctag agcctgtttt gcacgttcac cggcatcaac
      661 gttttctttt cggatccgcc gcataaccag tgaaacagca ttgctgtcac ttggtcgtgg
      721 cagcccggac cgacgatgaa gcatgtttag ctggcccaaa tgttgctgga tggtttttac
      781 tgccagaccg cgcgcctgaa gatatagaag ataatcgcga acatcttcag gttctgcggg
      841 aaaccatttc cggttattca acttgcacca tgccgcccac gaccggcaaa cggacagaag
      901 cattttccag gtatgctcag aaaacgcctg gcgatccctg aacatgtcca tcaggttctt
      961 gcgaacctca tcactcgttg catcgaccgg taatgcaggc aaattttggt gtacggtcag
     1021 taaattggac atggtggcgt acgagtaacg gcacggcgtc cacgacactc ccagcaccac
     1081 tgccggagcg cgagtctgcg ccccgcgata tatatatggt gcgccgtatc cctccgccgc
     1141 ggcggagccg cctcgcgcag agcgacgcgt gcggccgcag gaacccctag tgatggagtt
     1201 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     1261 acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac atgtgagcaa
     1321 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1381 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1441 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1501 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1561 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1621 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1681 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1741 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1801 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1861 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1921 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1981 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2041 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2101 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2161 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2221 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2281 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2341 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2401 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2461 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2521 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2581 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2641 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2701 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2761 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2821 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2881 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2941 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3001 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3061 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3121 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     3181 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     3241 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     3301 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     3361 tgagagtgca ccataaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
     3421 gttaaatcag ctcatttttt aaccaataga ccgaaatcgg caaaatccct tataaatcaa
     3481 aagaatagcc cgagatagag ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3541 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3601 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3661 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3721 aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct ggcaagtgta gcggtcacgc
     3781 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt
     3841 gctttgacgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     3901 gcgcccctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
     3961 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
     4021 ggccaactcc atcactaggg gttcctgcgg ccgctcggtc cgcacgtgcc acaccggtct
     4081 ccagatcttt aaaaaacctc ccacatctcc ccctgaacct gaaacataaa atgaatgcaa
     4141 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4201 caaatttcac aaataaagct agatggttac aaataaagca atagcatcac aaatttcaca
     4261 aataaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc agcgggcaag
     4321 gcaggcggcg atgagttccg ccgtggcaag aactaaccag gatttataca aggaggagaa
     4381 aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca
     4441 tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc
     4501 cagaggttga ttatctcgag tcatcagaat tcttacttgt catcgtcatc cttgtagtcg
     4561 atgtcatgat ctttataatc accgtcatgg tctttgtagt ctgtacaact ggtctcttta
     4621 agatcctcca ttggtagttt gtagctcatt atgaaggaag gaggtggtgc tggacctggg
     4681 ctaactggcc cctgcttctt tgcacagttt atacagatgt aatcttcatt ttcagccatt
     4741 tctggagata cacccacaca aacttgatga aaccactcat cacagccacc atcacattgt
     4801 acccagtcta ccttgtcctt gcagggcctt tggcagttct gtgctgcgca cacagcattc
     4861 tcatcatcag actcctctgc tcctgaccag tcatactttg aggggatgtc cagcaccttt
     4921 ttctctcttt tcttttcagt gctctctttc acaagttcaa ctttggctgc agcagccttc
     4981 tccttctttt tctttctctc ttcttctttt gctagtttct tggccagttt attcagctcc
     5041 tttgatttgt ctgcacctaa ttttaatttc ttctttctag gtttgtccat tttctttaac
     5101 tccttggact tctgttttcc ttctccaaaa agttgctcta ccttttctag cttccgtttc
     5161 cgtttcttct ctgaagagtc ctttcctttc acttttagtg gtttctcttc catgctgtca
     5221 tcctccatga tatgcaagaa tctgtcttca gagggtgggt gtgtggcctg caaaatccgc
     5281 catatgtgtt gagtctcgtc cagagatact tccaggagat ctccaaccat cataagttct
     5341 tccagttgtg ccttagctcc aggtgacaac tccagcactg gaggttccaa acttcggggc
     5401 accaaagggc tcttccgagg ttgtttcctt ggggtgctag aaccttgaga acaactctta
     5461 gaagcagaag aataagcatg ctctgcacaa aatgaaggtg ctgtccacat gtgggtcacc
     5521 acctcctcgg atttgatggg aatctcttca tcacaaaaaa gattgggttc aagactacta
     5581 gaggacttca cactggctgt gtccttcatg tcgtagccat atgtctctcg aatgtcttca
     5641 tcagagtctg tttcttcatc atcatagtcc attgtttgtc gaggagaaga tgacacactg
     5701 ctcaccaccc ggttaaaagc agactgctgg aaactaggta agtgtccctg taagtctgga
     5761 ttggcagctg ctttttggag ttctgcactg atgatctttt cagttttttc tcgagccgcc
     5821 tgttccacca tacgctggct caacacagat agtttggcca gggcagagga tagttcatct
     5881 gtggctagag cctgccgcgc tctatcttgc caactcatag cacgttctgt caaacactgc
     5941 agggcctctc cttcaggcaa ccgtacgggc aacttctgaa gggataccag gagtgacaga
     6001 atagtctcta gcctgggcct tcgagaccgc atacaaagag ggcaaaggaa ttttacttct
     6061 ttagcttgcc agctggatcc ttttttttgg gaacttgatt taggaagagg aacacagctg
     6121 ttatggaacc agtctttgca gagctcacac tgtagcataa acccactggc tgtcttgcgg
     6181 caaatgcaaa attttacttc ttctatgcgg tccaccattg tcatcttggc taggttggct
     6241 gctctgagag aatgcatggc ttcaatctct ttttgctccc gttctttgaa aactgccacc
     6301 accatggctg tatctctggt ttcctccaat ccttcctcca gatcactcag aggctccagg
     6361 tccagatcct tttctttttc tttttctatt agttctttta cttttttcct cctatttttg
     6421 ccactcccat atacaccaat gtcggtccgg gggctcagca cctgtggaga gaaaggcaaa
     6481 gtggatgtca gtaagaccaa taggtgccta tcagaaacgc aagagtcttc tctgtctcga
     6541 caagcccagt ttcataaatt gctataaagt atcctatacg aagttatact agttattggt
     6601 ctccttaaac ctgtcttgta accttgatac ttac
//
