LOCUS       sequence-491854-        5295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for ASTN2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5295)
  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  JOURNAL   Unpublished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 5295)
  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            "Unpublished"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'-ASTN2
FEATURES             Location/Qualifiers
     source          1..5295
                     /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        187..398
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             411..679
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      707..716
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   2558..2698
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2558..2687
                     /label=AAV2 ITR
     rep_origin      2773..3228
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2860..2879)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3070..3091
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3245..3264)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3364..3386
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3424..3442)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3510..3614
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3615..3683,3684..4475)
                     /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(3833..3852)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4646..5234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5135..5154
                     /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 gcgtgtcgac tctagaggat ctgcgatcgc tccggtgccc
      181 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
      241 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
      301 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
      361 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
      421 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
      481 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
      541 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
      601 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
      661 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac cgtacggcca ccatggccgc
      721 cgccggcgcc cggctcagcc ccggccccgg ctcggggctc cgggggcggc cgaggctctg
      781 cttccacccg gggccgccgc cactgctgcc gctgctgctg ctgttcctgc tcctgctgcc
      841 gccgccgccg ctgctggccg gcgccaccgc cgctgcctcg cgggagcccg acagcccgtg
      901 ccggctgaag accgtcacgg tgtccacact gcccgccctg cgggagagcg acatcggctg
      961 gagcggcgcc cgcgccgggg ccggggctgg gaccggggcc ggagccgccg ccgccgccgc
     1021 gtccccgggc tctcctggct ctgccggcac cgccgccgag tcgcgcctcc tgctctttgt
     1081 gcgtaacgag ctgccggggc gcatcgcggt gcaggacgac ctggacaaca ccgagctgcc
     1141 cttcttcacc ctggagatgt ctggcacagc ggcggacatc tcgctggtgc actggagaca
     1201 gcagtggctg gagaatggca ccttgtactt ccacgtctcc atgagcagct ccgggcagct
     1261 ggcccaagcc accgccccca ctctccagga gccctcggag attgttgagg agcagatgca
     1321 catcctccac atttctgtga tgggtggcct catcgcgctg ctgctgctgc tgctggtgtt
     1381 caccgtggcg ctgtacgccc agcgacgttg gcagaagcgt cgccgcatcc cccagaagag
     1441 cgcaagcaca gaagccactc atgagatcca ctacatccca tctgtgctgc tgggtcccca
     1501 ggcgcgggag agcttccgtt catcccggct gcaaacccac aattccgtca ttggcgtgcc
     1561 catccgggag actcccatcc tggatgacta tgactgtgag gaggatgagg agccacctag
     1621 gcgggccaac catgtctccc gcgaggacga gtttggcagc caggtgaccc acactctgga
     1681 cagtctggga catccagggg aagagaaggt ggactttgag aagaaagcag cagctgaggc
     1741 gactcaggag acagtggagt ccctgatgca gaagttcaag gagagtttcc gcgctaacac
     1801 gcccatcgag atcggtcagc tgcaaccacc cctgcgcagc acatcggcag ggaagaggaa
     1861 gcggaggagc aagtctcgag gaggaatcag ctttgggaga gccaagggga cgtcgggctc
     1921 agaggcagac gatgaaactc agctgacatt ctacacggag cagtaccgca gtcgccgccg
     1981 cagcaaaggt ttgctgaaaa gcccagtgaa caagacagcc ctgacactga ttgctgtgag
     2041 ttcctgcatc ctggccatgg tgtgtggcag ccagatgtct tgtccactca ctgtgaaggt
     2101 gactctgcat gtgcccgagc acttcatagc agatggaagc agcttcgtgg tgagtgaagg
     2161 gagctacctg gacatctccg actggttaaa cccagccaag ctttccctgt attaccagat
     2221 caatgccacc tccccatggg tgagggacct ctgtggacaa aggacgacag atgcctgtga
     2281 gcagctctgc gacccagaaa ccggagagtg cagctgtcat gaaggctatg cccctgaccc
     2341 tgttcacaga cacctgtgtg tgcgcagtga ctggggacag agtgaaggac cttggcccta
     2401 cacgacactt gagaggggct atgatctggt gacaggtaag tatcaaggtt acaagacagg
     2461 tttaaggaga ccaataagct attattcgta taggatactt tatacgaagt tattaagtta
     2521 tggagaccgg tgtggcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     2581 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     2641 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     2701 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     2761 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2821 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     2881 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2941 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     3001 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3061 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     3121 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     3181 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     3241 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3301 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3361 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3421 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3481 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3541 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3601 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3661 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3721 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3781 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3841 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3901 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3961 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4021 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4081 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4141 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4201 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4261 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4321 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4381 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4441 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4501 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4561 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4621 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4681 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4741 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4801 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4861 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4921 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4981 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5041 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5101 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5161 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5221 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5281 cttttgctca catgt
//
