LOCUS       sequence-476886-        5630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for KANK1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5630)
  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  JOURNAL   Unpublished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 5630)
  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            "Unpublished"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' KANK1
FEATURES             Location/Qualifiers
     source          1..5630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(77..103)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             140..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(421..632)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     repeat_region   677..806
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(868..1456)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(948..967)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1627..2418,2419..2487))
                     /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     2250..2269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2488..2592)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2660..2678
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2716..2738)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2838..2857
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2874..3329)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3011..3032)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3223..3242
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3404..3544
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3415..3544
                     /label=AAV2 ITR
ORIGIN
        1 tgtcctggtc tccactgaga atatcacctg cttttcctga ggcactgccg ttaacctttg
       61 tggtgtgagc agatccagca tagtcaggta cgtcataagg gtacatggtg gccgtacggt
      121 aggcgccggt cacagcttgg atctgtaacg gcgcagaaca gaaaacgaaa caaagacgta
      181 gagttgagca agcagggtca ggcaaagcgt ggagagccgg ctgagtctag gtaggctcca
      241 agggagcgcc ggacaaaggc ccggtctcga cctgagcttt aaacttacct agacggcgga
      301 cgcagttcag gaggcaccac aggcgggagg cggcagaacg cgactcaacc ggcgtggatg
      361 gcggcctcag gtagggcggc gggcgcgtga aggagagatg cgagcccctc gaagcttcag
      421 ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta ctgcacttat
      481 atacggttct cccccaccct cgggaaaaag gcggagccag tacacgacat cactttccca
      541 gtttaccccg cgccaccttc tctaggcacc cgttcaattg ccgacccctc cccccaactt
      601 ctcggggact gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc gatcgcagat
      661 ccacgcgtgc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
      721 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag
      781 cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa
      841 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      901 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      961 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1021 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1081 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1141 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1201 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1261 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     1321 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1381 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1441 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1501 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1561 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1621 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1681 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1741 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1801 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1861 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1921 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1981 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     2041 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     2101 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     2161 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     2221 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2281 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     2341 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2401 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2461 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2521 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2581 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     2641 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     2701 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     2761 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     2821 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt
     2881 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2941 ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt
     3001 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3061 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag
     3121 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3181 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3241 agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3301 cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa
     3361 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc
     3421 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
     3481 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
     3541 tcctgcggcc gctcggtccg cacgtgccac accggtctcc ataacttaat aacttcgtat
     3601 aaagtatcct atacgaataa tagcttattg gtctccttaa acctgtcttg taaccttgat
     3661 acttacctgt tccaaatctg tgctagtgtc ctggtctgca gtacgaggca ctgccatgac
     3721 ggcagcttcc acctggctaa cagcctcagt gttcacgccc cgggaggcgc actcctttgg
     3781 agagcagacg gtcacgtcaa cagaacaatc tccacaaccg atagaccgca cttctttgag
     3841 attcaagttt gtcttgagga gtgtgaggtc gttcacagac tcctctgtgt tgctgcctgt
     3901 ttcgcagacg ctgacttcca cactcacact cttgtcacac atttccacag acctcccagc
     3961 acatcggtca tgcatttcca ccctctcctt aacaatccac caattcatag gcagctcagg
     4021 ccctacgact ttattcttac attcaggctg gcaggagatg cctacactgt ttgtttccac
     4081 ggaggtccca acacacgtgt ccaccaggtc catgtgactg ccgaccattt ggtctctggt
     4141 ctgcaccact gcctccacca ccttactgaa aacaagcggc tgggccatcg tggctttgtc
     4201 aacctttttc ctcgatccag cagcctgcag ctcttgtttc agtttagtca tctcccggtc
     4261 atgggtggtt tctctaagct gtacttctag gcgatagatc ttttccttca aggcttctat
     4321 ggtctgctgt tggagctcaa tttctttgtc agcttcagtc atcactccaa gcatggcctc
     4381 tgtcacgctg accccacaat ttctcatctc aacaagtgtc cctacagctg catccttaca
     4441 ggacctggag cctctgtggt acacgacgat gtcgttcatg ttctcctcgg cacccacagc
     4501 cacagaccgg cactctccat tctccctgag ctctgaggag gcctcacagc tgctgtcctg
     4561 gatcttctgc tccagggctt gcatgtctgc tgtaagttgc cggaactcct ttatcctctg
     4621 cgtgctctgt tctacggtct ccatttcttc ctcctcatag tcaatgtata attccccgcc
     4681 acttcttcgg gcccgggaga gctgttccag ctgggaggcg ttccccgcac tataggaccg
     4741 cttcctcaca ccacagacat tgatctggga tgcagccctt tggtttttca gctgtgagac
     4801 caactgcctt ttctcttctt gcaagacaga gatctttacc tggagcacag ggatggttcg
     4861 cacctgctcc tccagctcct tcaggcgttt cagagcaatg gccatctgct cgcggatgtg
     4921 ctgcaggtgc atggggctca cgttggtcac tggggtggag atccctgagc tcagggggct
     4981 gtggcggatg gagctcccca tggaggaagt ggtgggagca gctggggcat agctaccata
     5041 atccccatta ccttggtatc cattctgaag ctggtgcttg gcaggattgt ggtttccaga
     5101 acccacaaaa gaagggaggg agcttgtggt gcccatgcct ccaaaactgg ccagcctggg
     5161 ccttctgaac tcacccggtg tcatctgcat ggtggctctc tcctgttcca gtcttctccg
     5221 ggtctccatc agtgtcttgg tgacatgaag gttatgcttt gggagttgtg gtgagggagg
     5281 tggcagctgt cgattttctg ggatggtaag aaaagggagt gaggtctcca gagggggagg
     5341 tggctttgag attggagttg atgtaacttg acttctggct atgaggaagt tggggcactg
     5401 cttgttgtca tcactgttgg aggatgagag ggattcagtg gaagtccata taccttgctg
     5461 accagatgtg gtcctgggtt ctgggcatgg cacggacggc ttccgcctct tctggatgtt
     5521 cagtcttttg atggtatttc ccttctgtat gtcatccaca tatttgagga aatctaagtc
     5581 tagttgataa ccataggggg tctccacaaa gtaagggtct ttctgttcct
//
