LOCUS       sequence-476874-        5948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' AAVLINK plasmid for KDM5B expression .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5948)
  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  JOURNAL   Unpublished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 5948)
  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            "Unpublished"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' KDM5B
FEATURES             Location/Qualifiers
     source          1..5948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1576..1641
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1706..1726)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2119..2259
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2119..2248
                     /label=AAV2 ITR
     rep_origin      2334..2789
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2421..2440)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2631..2652
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2806..2825)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2925..2947
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2985..3003)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3071..3175
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3176..3244,3245..4036)
                     /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(3394..3413)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4207..4795
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4696..4715
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4857..4986
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             join(5133..5948,1..216)
                     /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"
ORIGIN
        1 tacggcgcta aggatgactc tggtcagaga tacctggcct ggtctggaca cagtgcccgt
       61 gtcggagccg cgcgagatat ggcccgcgct ggagtttcaa taccggagat catgcaagct
      121 ggtggctgga ccaatgtaaa tattgtaatg aactatatcc gtaacctgga tagtgaaaca
      181 ggggcaatgg tgcgcctgct ggaagatggc gattaggtaa gtatcaaggt tacaagacag
      241 gtttaaggag accaataact agtataactt cgtataggat actttatagc aatttatgaa
      301 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      361 tgacatccac tttgcctttc tctccacagg gaaatggaag ccttgcagtc tctcagactc
      421 gccaatgaag ggaaattgct gtcgcctctc caagatgtgg atataaaaat ctgcctatgt
      481 cagaaggccc cagctgcccc tatgattcaa tgtgaactct gcagggatgc tttccacacc
      541 agttgtgtgg cggtacccag tatttcacag ggcctgcgaa tctggctttg tccccattgt
      601 cggaggtcag agaaacctcc attagagaaa attctgcccc tgctcgcctc ccttcagcgt
      661 atccgagttc gccttcctga gggagatgca cttcgatata tgattgaaag aaccgtgaac
      721 tggcagcaca gagcccagca actgctttcg tcagggaatc ttaaatttgt gcaagatcga
      781 gtgggctcag gactgttata tagcagatgg caagcctcag caggacaggt gtcagacaca
      841 aacaaggtat ctcaacctcc tggcacaaca tcattttctt tgcctgatga ctgggacaac
      901 agaacctcat atttgcactc ccccttctca actggacgaa gttgtatccc cctccatggt
      961 gttagtccag aagtgaatga actattgatg gaagcccagc tgctccaggt atcccttcct
     1021 gaaattcagg aactttacca gactttactt gcaaagccaa gccctgctca gcagactgac
     1081 cgaagctcac cagtgagacc cagcagtgag aagaatgact gttgccgagg gaagcgagat
     1141 ggaattaaca gtcttgagag aaaactgaag agacgcctgg aaagagaggg cctctccagt
     1201 gagcggtggg aacgagttaa gaaaatgcgg acccccaaaa agaagaaaat caaactgagc
     1261 caccccaagg acatgaacaa tttcaagtta gagagagagc gtagctatga attagttcgt
     1321 tctgctgaaa ctcattccct gccctcagac acatcctatt ccgaacagga agactctgag
     1381 gatgaagatg ccatctgccc agctgtgagc tgcctgcagc cagaaggaga tgaggtggac
     1441 tgggtccagt gtgatggcag ctgcaatcag tggtttcatc aggtctgtgt tggtgtctcc
     1501 ccagagatgg cagagaaaga agactacatc tgtgtgcgct gtactgtgaa ggacgcacca
     1561 agccgaaagt gtacagacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     1621 tacaaggatg acgatgacaa gtaactcgag ataatcaacc tctggattac aaaatttgtg
     1681 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1741 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1801 aatcctggtt agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     1861 caggggctcg gctgttgggc actgacaatt ccgtggtgtt tatttgtgaa atttgtgatg
     1921 ctattgcttt atttgtaacc atctagcttt atttgtgaaa tttgtgatgc tattgcttta
     1981 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2041 tttcaggttc agggggagat gtgggaggtt ttttaaagat ctggagaccg gtgtggcacg
     2101 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     2161 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     2221 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     2281 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     2341 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2401 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2461 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2521 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     2581 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2641 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     2701 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2761 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     2821 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     2881 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     2941 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3001 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     3061 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3121 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3181 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3241 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3301 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3361 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3421 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3481 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3541 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3601 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3661 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3721 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3781 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3841 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3901 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3961 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4021 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     4081 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4141 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4201 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4261 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4321 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4381 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4441 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4501 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4561 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4621 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4681 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4741 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4801 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg
     4861 caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc
     4921 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
     4981 gttcctgcgg ccgcacgcgt cgctctgcgc gaggcggctc cgccgcggcg gagggatacg
     5041 gcgcaccata tatatatcgc ggggcgcaga ctcgcgctcc ggcagtggtg ctgggagtgt
     5101 cgtggacgcc gtgccgttac tcgtacgcca ccatgtccaa tttactgacc gtacaccaaa
     5161 atttgcctgc attaccggtc gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca
     5221 tgttcaggga tcgccaggcg ttttctgagc atacctggaa aatgcttctg tccgtttgcc
     5281 ggtcgtgggc ggcatggtgc aagttgaata accggaaatg gtttcccgca gaacctgaag
     5341 atgttcgcga ttatcttcta tatcttcagg cgcgcggtct ggcagtaaaa accatccagc
     5401 aacatttggg ccagctaaac atgcttcatc gtcggtccgg gctgccacga ccaagtgaca
     5461 gcaatgctgt ttcactggtt atgcggcgga tccgaaaaga aaacgttgat gccggtgaac
     5521 gtgcaaaaca ggctctagcg ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg
     5581 aaaatagcga tcgctgccag gatatacgta atctggcatt tctggggatt gcttataaca
     5641 ccctgttacg tatagccgaa attgccagga tcagggttaa agatatctca cgtactgacg
     5701 gtgggagaat gttaatccat attggcagaa cgaaaacgct ggttagcacc gcaggtgtag
     5761 agaaggcact tagcctgggg gtaactaaac tggtcgagcg atggatttcc gtctctggtg
     5821 tagctgatga tccgaataac tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc
     5881 catctgccac cagccagcta tcaactcgcg ccctggaagg gatttttgaa gcaactcatc
     5941 gattgatt
//
