LOCUS       sequence-476864-        6849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' AAVLINK plasmid for YTHDC2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6849)
  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  JOURNAL   Unpublished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 6849)
  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            "Unpublished"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' YTHDC2
FEATURES             Location/Qualifiers
     source          1..6849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2143..2208
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2285..2305)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2698..2838
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2698..2827
                     /label=AAV2 ITR
     rep_origin      2913..3368
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3000..3019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3210..3231
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3385..3404)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3504..3526
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3564..3582)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3650..3754
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3755..3823,3824..4615)
                     /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(3973..3992)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4786..5374
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5275..5294
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5436..5565
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             5712..6743
                     /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 agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc
       61 tccacagggc gatgtagacc tggaatttgt tttcgtctgt tcagtagact ccgattccag
      121 aatatgttgg aatttcagac tccggaactt ttgagaatgc cattacagga actttgctta
      181 cataccaagc tgttagcccc agttaattgt cccattgctg attttcttat gaaagctcct
      241 gaacctccac cagctttaat tgtaagaaat gctgtacaaa tgcttaagac aatagatgca
      301 atggatacat gggaagatct gactgaactt gggtatcatt tggctgactt gccagtagaa
      361 ccacatcttg gtaaaatggt cttgtgtgct gttgttttaa agtgtctgga ccccatcctt
      421 acaattgctt gcacactagc ttatcgagat ccttttgtac tacctactca ggcctctcaa
      481 aaacgtgcag ctatgctttg taggaaacgt tttactgcag gagctttcag tgaccatatg
      541 gcacttctca gagcattcca ggcctggcaa aaagcacgaa gtgatgggtg ggagcgagcc
      601 ttttgtgaaa agaattttct ttcacaggct actatggaaa taatcatagg catgagaaca
      661 cagttgcttg gtcaacttag agcatcaggt tttgttagag cacgaggtgg tggtgacatt
      721 cgggacgtta acacaaactc tgagaattgg gctgtcgtta aagctgcatt ggtggcaggc
      781 atgtatccta atttagtcca cgtggacaga gagaatctag tgttgacagg gccaaaggag
      841 aaaaaagtac gatttcatcc tgcttcagtt ctcagtcagc ctcaatataa aaagattcct
      901 ccagccaatg gtcaagctgc agcaattaag gcactgccca cagattggct tatttatgat
      961 gaaatgacca gagcccatag aatagctaat attagatgtt gttcagcagt gacgcctgtc
     1021 actatattgg tattctgtgg accagctaga ttggcaagta atgctcttca ggaaccttca
     1081 tcctttagag tggatggcat tcccaatgac agtagtgata gtgaaatgga ggacaaaact
     1141 acagctaatt tggcagcctt gaaacttgat gagtggctcc atttcacact ggagccagag
     1201 gcagctagtt tattgctgca gctcagacag aagtggcata gcttattttt acgccgaatg
     1261 agagctccat ctaaaccttg gtctcaagtt gatgaagcta ccataagagc aattatagct
     1321 gttttaagca ctgaagaaca gtctgcaggt ttacaacaac catctgggat tggccaaagg
     1381 ccaaggccta tgtcttcaga agagcttcct ttggcctcat cttggaggtc aaataatagt
     1441 aggaaaagtt cagcagatac tgaattttct gatgagtgta ctactgcaga aagagtactg
     1501 atgaaatctc catctccagc attacaccca cctcagaagt acaaagatag aggaatttta
     1561 catcctaaac gaggtactga ggaccgatca gatcagtctt ctctgaaatc tacagacagc
     1621 agtagttacc caagtccttg tgctagtcct tctcctccat cctcaggaaa gggctcaaaa
     1681 tctccttcgc caagaccaaa catgcctgtt cgatacttca taatgaagag tagcaatttg
     1741 agaaaccttg aaatttctca acagaagggt atctggtcta caactcctag taatgaacgg
     1801 aagctaaatc gagccttttg ggaaagcagc atagtttact tggtattttc tgttcaagga
     1861 tctggacatt tccagggatt ttctaggatg tcttctgaga ttggaaggga aaagagtcag
     1921 gactggggct ctgctggact aggaggagta tttaaggtgg agtggatacg aaaagaaagc
     1981 cttccctttc aatttgcaca ccatttactc aatccatgga atgacaacaa gaaagtgcag
     2041 ataagcaggg atgggcagga actagaacct caggttggtg aacagttgct ccagttatgg
     2101 gaacgtcttc ccttgggaga aaaaaacaca actgattgta cagactacaa agaccatgac
     2161 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta agaattctga
     2221 tgactcgaga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2281 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2341 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     2401 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2461 ctgacaattc cgtggtgttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2521 tctagcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2581 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     2641 tgggaggttt tttaaagatc tggagaccgg tgtggcacgt gcggaccgag cggccgcagg
     2701 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     2761 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     2821 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2881 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     2941 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     3001 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3061 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3121 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3181 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3241 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     3301 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3361 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3421 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3481 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3541 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3601 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3661 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3721 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3781 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3841 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3901 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3961 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4021 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4081 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4141 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4201 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4261 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4321 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4381 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4441 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4501 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4561 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4621 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4681 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4741 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4801 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4861 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4921 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4981 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5041 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5101 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5161 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5221 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5281 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5341 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5401 gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc
     5461 gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
     5521 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgtc
     5581 gctctgcgcg aggcggctcc gccgcggcgg agggatacgg cgcaccatat atatatcgcg
     5641 gggcgcagac tcgcgctccg gcagtggtgc tgggagtgtc gtggacgccg tgccgttact
     5701 cgtacgccac catgtccaat ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg
     5761 atgcaacgag tgatgaggtt cgcaagaacc tgatggacat gttcagggat cgccaggcgt
     5821 tttctgagca tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca
     5881 agttgaataa ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat tatcttctat
     5941 atcttcaggc gcgcggtctg gcagtaaaaa ccatccagca acatttgggc cagctaaaca
     6001 tgcttcatcg tcggtccggg ctgccacgac caagtgacag caatgctgtt tcactggtta
     6061 tgcggcggat ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt
     6121 tcgaacgcac tgatttcgac caggttcgtt cactcatgga aaatagcgat cgctgccagg
     6181 atatacgtaa tctggcattt ctggggattg cttataacac cctgttacgt atagccgaaa
     6241 ttgccaggat cagggttaaa gatatctcac gtactgacgg tgggagaatg ttaatccata
     6301 ttggcagaac gaaaacgctg gttagcaccg caggtgtaga gaaggcactt agcctggggg
     6361 taactaaact ggtcgagcga tggatttccg tctctggtgt agctgatgat ccgaataact
     6421 acctgttttg ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc agccagctat
     6481 caactcgcgc cctggaaggg atttttgaag caactcatcg attgatttac ggcgctaagg
     6541 atgactctgg tcagagatac ctggcctggt ctggacacag tgcccgtgtc ggagccgcgc
     6601 gagatatggc ccgcgctgga gtttcaatac cggagatcat gcaagctggt ggctggacca
     6661 atgtaaatat tgtaatgaac tatatccgta acctggatag tgaaacaggg gcaatggtgc
     6721 gcctgctgga agatggcgat taggtaagta tcaaggttac aagacaggtt taaggagacc
     6781 aataactagt ataacttcgt ataggatact ttatagcaat ttatgaaact gggcttgtcg
     6841 agacagaga
//
