LOCUS       sequence-476889-        5814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for YTHDC2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5814)
  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  JOURNAL   Unpublished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 5814)
  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            "Unpublished"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' YTHDC2
FEATURES             Location/Qualifiers
     source          1..5814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(169..195)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             232..500
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(513..724)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   769..898
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(960..1548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1040..1059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1719..2510,2511..2579))
                     /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     2342..2361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2580..2684)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2752..2770
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2808..2830)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2930..2949
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2966..3421)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3103..3124)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3315..3334
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3496..3636
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3507..3636
                     /label=AAV2 ITR
ORIGIN
        1 gcgctccagc gcgatattga ctgcgatctt cacctcctca tcaatgcgaa tgtccttcag
       61 ccccttggcc cggccgccgc ccccagggcc acaaggcgag gggccgccgc ctccgccacc
      121 gccaggagcc ggctgccgcg gggagacgct gctcggcctg gaagatccag catagtcagg
      181 tacgtcataa gggtacatgg tggccgtacg gtaggcgccg gtcacagctt ggatctgtaa
      241 cggcgcagaa cagaaaacga aacaaagacg tagagttgag caagcagggt caggcaaagc
      301 gtggagagcc ggctgagtct aggtaggctc caagggagcg ccggacaaag gcccggtctc
      361 gacctgagct ttaaacttac ctagacggcg gacgcagttc aggaggcacc acaggcggga
      421 ggcggcagaa cgcgactcaa ccggcgtgga tggcggcctc aggtagggcg gcgggcgcgt
      481 gaaggagaga tgcgagcccc tcgaagcttc agctgtgttc tggcggcaaa cccgttgcga
      541 aaaagaacgt tcacggcgac tactgcactt atatacggtt ctcccccacc ctcgggaaaa
      601 aggcggagcc agtacacgac atcactttcc cagtttaccc cgcgccacct tctctaggca
      661 cccgttcaat tgccgacccc tccccccaac ttctcgggga ctgtgggcga tgtgcgctct
      721 gcccactgac gggcaccgga gcgatcgcag atccacgcgt gcggccgcag gaacccctag
      781 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
      841 aggtcgcccg acgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggac
      901 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      961 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1021 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1081 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1141 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1201 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1261 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1321 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1381 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1441 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     1501 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1561 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     1621 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     1681 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     1741 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     1801 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     1861 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     1921 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     1981 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2041 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2101 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2161 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     2221 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     2281 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     2341 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     2401 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     2461 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     2521 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     2581 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     2641 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     2701 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     2761 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     2821 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     2881 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     2941 cagattgtac tgagagtgca ccataaaatt gtaaacgtta atattttgtt aaaattcgcg
     3001 ttaaattttt gttaaatcag ctcatttttt aaccaataga ccgaaatcgg caaaatccct
     3061 tataaatcaa aagaatagcc cgagatagag ttgagtgttg ttccagtttg gaacaagagt
     3121 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3181 ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca
     3241 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3301 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctaaggcgct ggcaagtgta
     3361 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3421 tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     3481 accgcatcag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
     3541 caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
     3601 gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcggtc cgcacgtgcc
     3661 acaccggtct ccataactta ataacttcgt ataaagtatc ctatacgaat aatagcttat
     3721 tggtctcctt aaacctgtct tgtaaccttg atacttacct gccctgcctt tccgctgtat
     3781 ggcactagct ttggaaatcc ataccatttt taacattgta acaaaattca gagcatcaaa
     3841 ggatttctct ttcaccttac cagaatcaat aacaaagaca acatcattga ctgtgatgct
     3901 ggtttcagca atattggtgg aaagaattat ttttcgaaca cctgcaggtg ggttttttaa
     3961 tactttcttt tgatcggatg tttgcatatt tgaatgaagc ataaagactt ggtatctatg
     4021 tgtactgtca gcaaaccgct tgtcatcaaa caggatgcga tctctcagtc caacaatttc
     4081 gtcatatcca ggcagaaaaa ttagtactgc accagcatca caactatggc agatattgta
     4141 tagaagatgc atgatcaaat ccaagtctac tttttcatca tcgaaactat gatgataagc
     4201 ttccaggagc tctctgtctt cagcactgag gtcacttcca tttgtttgaa ccagagaact
     4261 ttcatctaga tttccaaatt ccagtgtagc actgtaagat tctagaagat ccacaatttc
     4321 agtctgccca aagtgtttag cccaatccaa tgccatccag ccatttgatg ctttactatg
     4381 gacattggct cccatactga ttaactgttc tacttgactt gcaaagccac gtcctgcagc
     4441 aaccatcaga gctgttgcac tggtttcact atgtctgtaa tcaacactaa cattttcagt
     4501 taaaatgaga tgaaagacct gagcaaaggc atcaatatct ttatgtagcc atatatcaga
     4561 aaggcaagca tccatttcct ttattaacca tggttcaagg caattcacat ctttttcagt
     4621 cagctgactg aagacagcat caccaccatc atccagtaag tcatactcat cagtcacatt
     4681 tagaacagtt ctctgcctct gagattcagg cttgaaacta ttttcttgag ctgagtacca
     4741 ttctgtaagt gtggtttgtt gtttctcttc ttgctgtttt tcctttttat attttaacat
     4801 ttctttgttt gtatatccag ttgttcttaa aatatcttcc agaaacattt cttttacttc
     4861 aaatggtctt ccctgtatat atatcactgg acaacttcca aaatatctta taaagagatt
     4921 tacatccaag gcagcactag aaagaattag tttcaaagtt gggtgctttt gcaacaaatc
     4981 tcttaacttt gtaagtaaaa aatcactaaa tcgatccctt tcatgcactt catccacgat
     5041 aacatgtgtc acagtcgaca acgtactatc tcctgccatc aatgtacgaa gcaatacccc
     5101 attagtacaa aatgtcagaa gtgtctttgg agaaaccctg ctttctaatc ggatctgata
     5161 accaattgtt tgaccaatcc tttcccgtct ctctgcggca actctttcag ccacagcgat
     5221 agctgccaat cgtcttggtt gagtacaaaa tatacggcag gggataccat ttttaaagca
     5281 atcatctaaa aggaactgag gaatctgtgt ggtctttcca gacccagttt ctcctacaat
     5341 caaaactact ttattttcct taattatttt aacaatttct tcctgtttct caaacactgg
     5401 taaagactgc ctaaaagaat caaattcgga ttctcctctt ttcactggaa tctgaggtat
     5461 gccattgttg agtcgcccac ttgtcttgct catttcccgg ttttcagctt caactgcaaa
     5521 cacatttcct ctttctgttt taggcagaag ttctgtacgc tctttattgg tgacaggaaa
     5581 tctttgaatt aggctcctaa cagcatgttt tgtattatga gtcaaattac aggtcatcat
     5641 tgcatgagct gtttctgatc catctttctt cttcacagtt aggtatctat ttgctccctt
     5701 tcctttactt ttagagacca aaccaagaga ctgactgagt cgatgaataa aggctctttc
     5761 agtactggtc aaagaagaag gaaattccat ttctctctgg tccccgtatc ggaa
//
