LOCUS       sequence-497521-        6060 bp    DNA     circular SYN 15-JAN-2026
DEFINITION  5' AAVLINK plasmid for KIAA0232 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6060)
  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  JOURNAL   Unpublished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 6060)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 15, 2026 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            "Unpublished"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'-KIAA0232
FEATURES             Location/Qualifiers
     source          1..6060
                     /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"
     repeat_region   3323..3463
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3323..3452
                     /label=AAV2 ITR
     rep_origin      3538..3993
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3625..3644)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3835..3856
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4010..4029)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4129..4151
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4189..4207)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4275..4379
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4380..4448,4449..5240)
                     /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(4598..4617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5411..5999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5900..5919
                     /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 ggatcttacc ctatctgtac agttgttgtg gatggtttgc catctgaaag
      781 ctcctcaagt tcttatccag gccctgtgtc tgtttctgaa atgtctctgc ttcatgcttt
      841 gggtccagtg cagacctggc tgggacaaga gctcgagaaa tgtggcattg atgccatgat
      901 ttacactcgg tatgtcctca gtcttctgct gcatgacagc tatgactacg acctgcagga
      961 acaggagaat gacatcttcc tgggctggga aaaaggagct tataagaaat ggggaaagag
     1021 taagaaaaaa tgttcagatc taactctaga agaaatgaaa aaacaggctg ctgtccagtg
     1081 tcttcgatct gcttctgatg aaagctctgg tatcgagact ttagtggagg agctctgctc
     1141 cagactgaaa gaccttcaga gtaagcaaga agagaagatt cacaaaaagt tagaggggtc
     1201 tccctctcca gaggcagaat tatcccctcc agcaaaggat caagtggaaa tgtactatga
     1261 agcatttcca ccactttctg agaaaccagt ttgcctgcaa gaaatcatga ctgtgtggaa
     1321 caagtctaaa gtctgttctt actctagctc ttcttcatca tccacagccc caccagctag
     1381 cacagatact tcctctccta aggactgcaa cagtgaaagt gaagtcacca aggaaagaag
     1441 cagtgaagta cccaccactg tgcatgagaa aacccagagc aaaagcaaaa acgagaagga
     1501 aaacaaattt agtaatggca caattgaaga aaagcctgct ttgtacaaaa agcaaatccg
     1561 acataaacct gaaggaaaga ttcgccctcg ctcgtggtct tctggctcca gtgaagcagg
     1621 ctcaagttcc agtgggaatc agggagaatt aaaagcatcc atgaagtatg ttaaagtaag
     1681 acacaaggca cgagagattc gaaacaaaaa agggcggaat gggcaaagca ggctttcttt
     1741 gaagcacggt gaaaaggctg aaaggaacat tcatactgga agtagtagca gtagcagcag
     1801 tggttctgtc aaacagctgt gcaagcgggg taagagacct ttaaaagaaa tagggagaaa
     1861 agatcctggg agcactgaag gaaaagacct gtacatggag aatagaaagg acacagagta
     1921 taaagaggag cccttgtggt acaccgagcc aattgctgaa tattttgttc ctctgagcag
     1981 aaaaagtaaa ctagagacca cataccgaaa cagacaggat acaagtgatc tgacatcaga
     2041 ggcagtggaa gaattgtctg aatcagtgca tggtctttgt atcagcaaca ataatcttca
     2101 taaaacatac ctcgcagcag gtactttcat tgatggtcat tttgtagaaa tgcctgcagt
     2161 tataaatgag gatattgacc tcactgggac ctcattatgt tctctaccag aggacaataa
     2221 atacctggat gatattcatc tatcagaatt aacgcacttc tatgaagtgg atattgatca
     2281 atccatgttg gatcctggtg cctcagaaac aatgcaagga gaaagtcgga ttttgaatat
     2341 gattcgacag aaaagcaaag agaacacaga ttttgaggca gaatgttgca tagtgttaga
     2401 tggtatggag ttgcaagggg aacgtgcaat atggacagat tctaccagct ccgtaggtgc
     2461 tgagggctta ttcctgcagg accttggcaa tctggctcag ttttgggagt gctgttcatc
     2521 cagctccggt gatgctgatg gggagagttt tggaggagac tctccagtta gactctctcc
     2581 catcttagac agcacagtgc tcaattcaca cctgcttgct ggcaatcaag agctcttttc
     2641 agatattaat gaaggatctg gtataaactc ttgtttttca gtgtttgaag tgcaatgcag
     2701 taattctgtt ttaccatttt cttttgaaac actcaacttg ggaaatgaaa atacagattc
     2761 tagtgctaat atgcttggga aaacacagtc tagattgcta atatggacca aaaatagtgc
     2821 ctttgaagaa aatgaacact gttctaatct ttcaacaaga acttgtagtc catggtccca
     2881 ttcagaagaa acacgttcag acaatgaaac attaaatatt cagtttgaag aatccacaca
     2941 gtttaatgcc gaagatatta attatgtagt tcctagagtc tcgtcaaatt atgtagatga
     3001 agaacttcta gattttttgc aagatgaaac ttgccagcaa aacagtagaa ctttaggtga
     3061 gattcctaca ttagttttca aaaaaacatc taaactagaa tccgtctgtg gtattcagct
     3121 agaacaaaaa acagaaaaca aaaattttga aactacacaa gtatgtaatg aaagtccaca
     3181 tggagatggc tacagctcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     3241 aagctattat tcgtatagga tactttatac gaagttatta agttatggag accggtgtgg
     3301 cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3361 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3421 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     3481 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3541 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3601 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3661 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3721 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     3781 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3841 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     3901 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3961 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     4021 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4081 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4141 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4201 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4261 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4321 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4381 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4441 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4501 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4561 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4621 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4681 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4741 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4801 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4861 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4921 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4981 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5041 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5101 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5161 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5221 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5281 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5341 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5401 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5461 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5521 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5581 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5641 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5701 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5761 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5821 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5881 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5941 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6001 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
