LOCUS       sequence-491430-        6755 bp    DNA     circular SYN 03-NOV-2025
DEFINITION  5' AAVLINK plasmid for SHANK2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6755)
  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  JOURNAL   Unpublished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 6755)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 3, 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            "Unpublished"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'-SHANK2
FEATURES             Location/Qualifiers
     source          1..6755
                     /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"
     CDS             716..742
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     repeat_region   4018..4158
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4018..4147
                     /label=AAV2 ITR
     rep_origin      4233..4688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4320..4339)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4530..4551
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4705..4724)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4824..4846
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4884..4902)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4970..5074
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5075..5143,5144..5935)
                     /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(5293..5312)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6106..6694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6595..6614
                     /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 ccatgtaccc
      721 ttatgacgta cctgactatg ctggatctat gccgcgcagc ccaacatcca gcgaggacga
      781 gatggcccag agcttctccg actactccgt ggggtcggag tcagacagct ccaaagaaga
      841 gaccatctat gacacgatcc gggccactgc ggagaagccg ggcggtgcca ggacggagga
      901 gagccagggc aacacgctgg tgatccgcgt ggtcatccat gacctgcagc agacgaaatg
      961 cattcgattt aacccggatg ccacagtgtg ggttgcaaag cagcggatcc tgtgtacatt
     1021 aacccagagt ttgaaagatg tcctgaacta cggcctgttc cagccggcca gcaatgggcg
     1081 tgacggcaag ttcctggatg aggagcggct cctgcgcgag tacccacagc ccgtgggtga
     1141 gggcgttcct tccctggagt ttcgatacaa gaagcgggtg tataaacaag ccagtctcga
     1201 tgagaaacag ttggccaagc tccacacgaa gaccaatctg aagaaatgca tggatcacat
     1261 tcagcatcgc ttggtggaga agatcaccaa gatgctggac cgaggcctgg atcccaattt
     1321 ccacgacccg gagaccggag agacccccct gaccttagcc gctcagctgg acgactctgt
     1381 ggaggtcatc aaagctctca aaaatggtgg agctcacctg gacttccgtg ccaaagatgg
     1441 gatgaccgcc ctacacaaag ctgcccgagc gaggaaccaa gttgccctga agaccctttt
     1501 agagcttggt gcatccccag attataaaga cagttacggc ctcaccccgc tgtatcacac
     1561 agccatcgtc ggaggtgatc cctactgctg cgagcttctc ctgcacgaac acgccactgt
     1621 gtgctgcaaa gatgagaacg gctggcacga gatccaccag gcctgcaggt acgggcacgt
     1681 gcagcacctg gagcacctgc tgttctacgg ggcagacatg agtgcccaga atgcctcggg
     1741 gaacacggcc ttgcacatct gcgccctcta caaccaggac agctgtgcaa gagtgcttct
     1801 gtttcgaggc ggaaataagg agttaaaaaa ctacaacagc cagactccat ttcaggtggc
     1861 cataatagca ggcaactttg agctggcaga atacatcaag aaccacaagg aaacagacat
     1921 tgtgcccttc cgagaggccc cggcgtactc caaccgccgg cggcggcccc ccaacacgct
     1981 ggccgccccc cgggtcctgc tgcgctccaa cagtgacaac aacctcaatg ccagcgctcc
     2041 cgactgggcc gtctgctcca cggccacctc gcaccgcagc ctgtcacccc agctgctgca
     2101 gcagatgccc agcaagcccg agggggccgc gaagaccatt gggagctacg tgcccgggcc
     2161 ccgcagccgg tccccatcgc tcaacaggct gggcggcgca ggcgaggacg gcaagaggcc
     2221 gcagcctctc tggcatgtcg ggtcgccttt tgctcttggt gccaacaagg actcactctc
     2281 ggccttcgag tacccggggc ccaagcggaa gctctacagt gccgtgcccg ggaggctctt
     2341 cgtcgctgtc aagccatacc aaccccaagt ggacggcgag atcccccttc accgcggtga
     2401 cagggtcaaa gttctgagca tcggtgaagg gggcttctgg gaaggcagcg cccgcggcca
     2461 catcggatgg tttccggcgg agtgcgtgga ggaggtccag tgtaagccca gggacagcca
     2521 ggcagaaacg cgcgctgacc gcagcaagaa gcttttcagg cactacaccg tgggctccta
     2581 tgacagcttc gacacttcca gtgactgcat tattgaggag aagacggtgg tcctgcagaa
     2641 aaaagacaat gagggctttg gattcgtgct tcgaggggcc aaagctgaca cacccattga
     2701 agaattcaca ccaacaccgg ctttcccagc cctacagtac ctggagtccg tggatgaagg
     2761 tggggtggcg tggcaagccg gactaaggac cggggacttc ttgattgagg ttaacaatga
     2821 gaatgttgtc aaagtcggcc acaggcaggt ggtgaacatg atccggcagg gagggaatca
     2881 cctggtcctt aaggtggtca cggtgaccag gaatctggac cccgacgaca ccgccaggaa
     2941 gaaagctccc ccgcctccaa agcgggcacc gaccacagcc ctcaccctgc gctccaagtc
     3001 catgacctcg gagctggagg agctcgtgga taaagcctcg gtccggaaga agaaggataa
     3061 acccgaggag atagtcccgg cctccaagcc ctcccgcgct gctgagaaca tggctgtgga
     3121 accgagggtg gcgaccatca agcagcggcc cagcagccgg tgcttcccgg cgggctcaga
     3181 catgaactct gtgtacgaac gccaaggaat cgccgtgatg acgcccactg ttcctgggag
     3241 cccaaaagcc ccgtttctgg gcatccctcg aggtacgatg cgaaggcaga aatcaataga
     3301 cagcagaatc tttctatcag gaataacaga ggaagagcgg cagtttctgg ctcctccaat
     3361 gctgaagttc accagaagcc tgtccatgcc ggacacctct gaggacatcc cccctccacc
     3421 gcagtctgtg cccccgtccc caccaccacc ttccccaacc acttacaact gccccaagtc
     3481 cccaactcca agagtctacg ggacgattaa gcctgcgttc aatcagaatt ctgccgccaa
     3541 ggtgtccccc gccaccaggt ccgacaccgt ggccaccatg atgagggaga aggggatgta
     3601 cttcaggaga gagctggacc gctactcctt ggactctgaa gacctctaca gtcggaatgc
     3661 cggcccgcaa gccaacttcc gcaacaagag aggccagatg ccagaaaacc catactcaga
     3721 ggtggggaag atcgccagca aagccgtcta cgtccccgcc aagcccgcca ggcggaaggg
     3781 gatgctggtg aagcagtcca acgtggagga cagccccgag aagacgtgct ccatccctat
     3841 cccgaccatc atcgtgaagg agccgtccac cagcagcagc ggcaagagca gccaggtaag
     3901 tatcaaggtt acaagacagg tttaaggaga ccaataagct attattcgta taggatactt
     3961 tatacgaagt tattaagtta tggagaccgg tgtggcacgt gcggaccgag cggccgcagg
     4021 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     4081 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     4141 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4201 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     4261 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     4321 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4381 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4441 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4501 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4561 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     4621 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     4681 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4741 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4801 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4861 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4921 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4981 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5041 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5101 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5161 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5221 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5281 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5341 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5401 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5461 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5521 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5581 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5641 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5701 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5761 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5821 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5881 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5941 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6001 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6061 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6121 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6181 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6241 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6301 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6361 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6421 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6481 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6541 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6601 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6661 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6721 gttcctggcc ttttgctggc cttttgctca catgt
//
