LOCUS       sequence-491796-        5770 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  5' AAVLINK plasmid for YEATS2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5770)
  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  JOURNAL   Unpublished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 5770)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 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            "Unpublished"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-HA-5'-YEATS2
FEATURES             Location/Qualifiers
     source          1..5770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..386
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             399..667
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             704..730
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      3248..3703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3335..3354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3545..3566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3720..3739)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3839..3861
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3899..3917)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3985..4089
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4090..4158,4159..4950)
                     /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(4308..4327)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5121..5709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5610..5629
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtggatct gcgatcgctc cggtgcccgt cagtgggcag
      181 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
      241 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      301 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      361 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
      421 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
      481 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
      541 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
      601 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
      661 acagatccaa gctgtgaccg gcgcctaccg tacggccacc atgtaccctt atgacgtacc
      721 tgactatgct ggatcttctg gaatcaagcg aaccatcaaa gaaaccgacc ctgattacga
      781 ggatgtatct gtggcccttc caaataagcg gcataaagca attgagaatt cagctcgaga
      841 tgctgctgtg cagaagattg agactattat caaagaacag tttgctcttg aaatgaagaa
      901 taaggaacat gaaattgaag tcattgacca gcgactgatt gaagcaagaa ggatgatgga
      961 taaactgcgt gcctgcattg tagcaaacta ctatgcttct gcaggtcttc taaaagtttc
     1021 tgagggatca aagacatgtg atacaatggt ttttaatcat cctgctatca agaaattttt
     1081 ggaatcacca tctaggtcat catctcctgc caatcagaga gcagaaacac catcagccaa
     1141 tcattcagaa agtgattctt tatctcagca caatgacttc ttatctgaca aagataataa
     1201 cagcaatatg gatatagagg aaagactctc aaacaacatg gagcagagac caagccgaaa
     1261 tactggaagg gatacttcta gaattactgg ctcccataaa acagaacagc ggaatgctga
     1321 tctcacagat gagacttcac gactttttgt aaagaaaaca atagtagtgg gcaatgtgtc
     1381 caagtatata cctccggata agagggaaga aaatgaccag tcaactcata agtggatggt
     1441 atatgtccga gggtcccgta gagaacccag cattaatcat tttgtcaaga aggtttggtt
     1501 cttccttcat cctagctata aaccaaatga ccttgtggaa gttagagagc ctccttttca
     1561 cctgaccaga agaggctggg gtgagtttcc cgtcagagtt caagttcatt ttaaggacag
     1621 ccagaacaag cggatagata tcatacataa tctgaagctg gatagaactt atactggctt
     1681 gcagactctt ggagcagaga cggtagtgga tgttgaactc catcgccatt ctctcggaga
     1741 agactgtatc tatcctcagt cctcggagtc tgacatctct gatgcccctc catctttgcc
     1801 tttgaccatt ccagccccag tgaaagcttc ttcaccaata aagcagtcac atgagccagt
     1861 acccgatacc tctgtggaga aaggattccc agctagcact gaagctgaac gacacactcc
     1921 gttttatgct ttgccatctt cattggaaag aacacccacc aaaatgacta catcccagaa
     1981 agttaccttt tgttcccatg gcaattcagc tttccagcca atagcatcaa gctgcaaaat
     2041 tgttccacaa agtcaggttc ctaatcctga gtcacctgga aaatccttcc agcccatcac
     2101 catgagctgc aagattgtgt caggttcccc aatatcaact ccaagcccat caccattgcc
     2161 tcgaaccccg acttccactc cagtccacgt gaagcaaggc actgccggct ctgttattaa
     2221 taatccttat gttatcatgg acaagcagcc ggggcaggtg attggagcca ccactcccag
     2281 tacaggaagt cctacaaaca agatctccac ggcttctcag gtctcccaag gaacaggttc
     2341 ccctgttcct aaaattcatg gaagtagttt tgtaacatct actgtcaagc aggaggattc
     2401 tttgtttgca tctatgccac ctctttgccc aattgggagt caccctaagg ttcaaagccc
     2461 caaacctata acaggaggac ttggagcttt cacaaaagtg atcatcaaac aggaacctgg
     2521 tgaagcccct cacgtgcccg caacaggagc tgccagccag tcaccactcc cgcagtatgt
     2581 gactgtgaaa gggggtcaca tgatagctgt gtcccctcaa aaacaggtca taactcctgg
     2641 agaagggatt gcccagtcag caaaggttca gccctccaag gttgtcgggg taccagttgg
     2701 gtctgcttta ccttcaacag tgaagcaggc tgtggcgatc agtggtggcc agatcctggt
     2761 agccaaggcc agctcttctg tctccaaagc agttgggcca aagcaagttg taacccaagg
     2821 agttgccaaa gcaattgtga gtggaggtgg aggaaccatt gttgctcagc cagtgcagac
     2881 cttaaccaag gcccaggtta ctgccgctgg tcctcagaag agtggatccc aggtaagtat
     2941 caaggttaca agacaggttt aaggagacca ataagctatt attcgtatag gatactttat
     3001 acgaagttat taagttatgg agaccggtgt ggcacgtgcg gaccgagcgg ccgcaggaac
     3061 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     3121 gaccaaaggt cgcccggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     3181 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3241 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3301 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     3361 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3421 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3481 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3541 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     3601 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     3661 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3721 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3781 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3841 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3901 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3961 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4021 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4081 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4141 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4201 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4261 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4321 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4381 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4441 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4501 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4561 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4621 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4681 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4741 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4801 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4861 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4921 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4981 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5041 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5101 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5161 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5221 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5281 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5341 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5401 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5461 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5521 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5581 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5641 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5701 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5761 gctcacatgt
//
