LOCUS       sequence-476861-        6645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' AAVLINK plasmid for CHD2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6645)
  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  JOURNAL   Unpublished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 6645)
  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            "Unpublished"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' CHD2
FEATURES             Location/Qualifiers
     source          1..6645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2053..2118
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2183..2203)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2596..2736
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2596..2725
                     /label=AAV2 ITR
     rep_origin      2811..3266
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2898..2917)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3108..3129
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3283..3302)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3402..3424
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3462..3480)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3548..3652
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3653..3721,3722..4513)
                     /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(3871..3890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4684..5272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5173..5192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5334..5463
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             5610..6641
                     /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 gtatcaaggt tacaagacag gtttaaggag accaataact agtataactt cgtataggat
       61 actttatagc aatttatgaa actgggcttg tcgagacaga gaagactctt gcgtttctga
      121 taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg gaaaaggaga
      181 ggtccaacaa tcaagatatc cggagttcag gttaatgtga aatccattat ccaacatgaa
      241 gaggagtttg agatgctgca taaatctatc cctgtggacc ctgaagaaaa aaaaaaatac
      301 tgcttaacct gtcgtgtcaa agctgcacat tttgatgtag agtggggggt ggaagatgat
      361 tctcgcctgt tgctggggat ttatgaacat ggctatggaa actgggagtt aattaaaaca
      421 gacccagagc ttaaattaac tgacaaaatt ctgccggtgg agacagataa aaagcctcag
      481 gggaagcagc tacagacccg agcggattac ttgttgaagc tgctcagaaa gggtctggag
      541 aagaaggggg ctgtgacagg tggggaagag gccaaattaa agaagcggaa gcctcgggta
      601 aagaaggaaa acaaagtgcc caggctgaaa gaggagcatg gaattgagct ttcatctcct
      661 aggcattcag ataatccatc agaagaggga gaagtgaaag atgatggctt ggaaaaaagt
      721 ccaatgaaaa aaaaacagaa gaagaaagag aacaaggaga acaaggagaa acaaatgagt
      781 tctaggaaag acaaagaagg ggacaaggaa agaaagaagt caaaagataa gaaagagaag
      841 cctaaaagtg gtgatgccaa atcttcgagt aaatcaaagc gatctcaggg tcctgtccat
      901 attacagcag gaagtgaacc tgtccccatt ggagaggatg aggatgatga tctggaccag
      961 gagacattca gcatatgtaa ggagaggatg aggcccgtga aaaaggcact gaaacagctc
     1021 gacaaacctg acaaggggct caacgtgcaa gaacagctgg aacacacccg gaactgcctg
     1081 ctgaaaatcg gagaccggat agccgagtgc cttaaagcct actcagatca ggagcacatc
     1141 aaactctgga ggaggaacct atggattttt gtttccaagt ttacagaatt tgatgctcga
     1201 aaactgcata agttatacaa gatggctcat aagaaaaggt ctcaagaaga agaggagcaa
     1261 aagaagaaag acgacgtgac tgggggtaag aaaccatttc gtccagaggc ctcaggctcc
     1321 agccgggact ctctgatatc tcagtcccat acctcacaca accttcaccc tcagaagcct
     1381 catttgcctg cctcccatgg cccacagatg catggacacc caagagataa ctacaatcac
     1441 cccaacaaga gacacttcag taatgcagat cgaggagact ggcagaggga aagaaagttc
     1501 aactatggtg gtggcaacaa caatccacca tggggaagcg acaggcacca tcagtatgag
     1561 cagcactggt acaaggacca ccattatggg gaccggcgac atatggatgc ccaccgttcc
     1621 ggaagctatc gacccaacaa catgtccaga aagaggcctt atgaccagta cagcagtgac
     1681 cgagaccacc ggggacacag agattattat gacaggcacc atcatgactc caagcggagg
     1741 agatccgatg aatttaggcc tcaaaattac caccagcagg atttccgacg aatgtctgat
     1801 caccgccccg ctatgggcta ccatggccag ggaccctcag accattaccg ctctttccac
     1861 acagataaac tgggggaata taaacagcct ctacccccat tgcaccctgc agtctcagat
     1921 cctcgctcac ccccttctca gaaatctcct cacgattcca agtcacccct ggatcatagg
     1981 tctcctttgg agagatcact agaacagaaa aacaacccag attataactg gaatgttcgg
     2041 aaaacatgta cagactacaa agaccatgac ggtgattata aagatcatga catcgactac
     2101 aaggatgacg atgacaagta actcgagata atcaacctct ggattacaaa atttgtgaaa
     2161 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2221 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2281 cctggttagt tcttgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2341 gggctcggct gttgggcact gacaattccg tggtgtttat ttgtgaaatt tgtgatgcta
     2401 ttgctttatt tgtaaccatc tagctttatt tgtgaaattt gtgatgctat tgctttattt
     2461 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2521 caggttcagg gggagatgtg ggaggttttt taaagatctg gagaccggtg tggcacgtgc
     2581 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2641 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2701 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     2761 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     2821 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2881 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2941 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3001 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     3061 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3121 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     3181 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3241 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     3301 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3361 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3421 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3481 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3541 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3601 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3661 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3721 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3781 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3841 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3901 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3961 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4021 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4081 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4141 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4201 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4261 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4321 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4381 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4441 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4501 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4561 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4621 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4681 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4741 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4801 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4861 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4921 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4981 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5041 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5101 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5161 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5221 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5281 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag
     5341 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc
     5401 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     5461 cctgcggccg cacgcgtcgc tctgcgcgag gcggctccgc cgcggcggag ggatacggcg
     5521 caccatatat atatcgcggg gcgcagactc gcgctccggc agtggtgctg ggagtgtcgt
     5581 ggacgccgtg ccgttactcg tacgccacca tgtccaattt actgaccgta caccaaaatt
     5641 tgcctgcatt accggtcgat gcaacgagtg atgaggttcg caagaacctg atggacatgt
     5701 tcagggatcg ccaggcgttt tctgagcata cctggaaaat gcttctgtcc gtttgccggt
     5761 cgtgggcggc atggtgcaag ttgaataacc ggaaatggtt tcccgcagaa cctgaagatg
     5821 ttcgcgatta tcttctatat cttcaggcgc gcggtctggc agtaaaaacc atccagcaac
     5881 atttgggcca gctaaacatg cttcatcgtc ggtccgggct gccacgacca agtgacagca
     5941 atgctgtttc actggttatg cggcggatcc gaaaagaaaa cgttgatgcc ggtgaacgtg
     6001 caaaacaggc tctagcgttc gaacgcactg atttcgacca ggttcgttca ctcatggaaa
     6061 atagcgatcg ctgccaggat atacgtaatc tggcatttct ggggattgct tataacaccc
     6121 tgttacgtat agccgaaatt gccaggatca gggttaaaga tatctcacgt actgacggtg
     6181 ggagaatgtt aatccatatt ggcagaacga aaacgctggt tagcaccgca ggtgtagaga
     6241 aggcacttag cctgggggta actaaactgg tcgagcgatg gatttccgtc tctggtgtag
     6301 ctgatgatcc gaataactac ctgttttgcc gggtcagaaa aaatggtgtt gccgcgccat
     6361 ctgccaccag ccagctatca actcgcgccc tggaagggat ttttgaagca actcatcgat
     6421 tgatttacgg cgctaaggat gactctggtc agagatacct ggcctggtct ggacacagtg
     6481 cccgtgtcgg agccgcgcga gatatggccc gcgctggagt ttcaataccg gagatcatgc
     6541 aagctggtgg ctggaccaat gtaaatattg taatgaacta tatccgtaac ctggatagtg
     6601 aaacaggggc aatggtgcgc ctgctggaag atggcgatta ggtaa
//
