LOCUS       sequence-479515-        7044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for ITSN1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7044)
  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  JOURNAL   Unpublished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 7044)
  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            "Unpublished"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' ITSN1
FEATURES             Location/Qualifiers
     source          1..7044
                     /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   4318..4447
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      4522..4977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4609..4628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4819..4840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4994..5013)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5113..5135
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5173..5191)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5259..5363
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5364..5432,5433..6224)
                     /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(5582..5601)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6395..6983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6884..6903
                     /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 ggatctgctc agtttccaac accttttggt ggcagcctgg atatctgggc
      781 cataactgta gaggaaagag cgaagcatga tcagcagttc catagtttaa agccaatatc
      841 tggattcatt actggtgatc aagctagaaa cttttttttt caatctgggt tacctcaacc
      901 tgttttagca cagatatggg cactagctga catgaataat gatggaagaa tggatcaagt
      961 ggagttttcc atagctatga aacttatcaa actgaagcta caaggatatc agctaccctc
     1021 tgcacttccc cctgtcatga aacagcaacc agttgctatt tctagcgcac cagcatttgg
     1081 tatgggaggt atcgccagca tgccaccgct tacagctgtt gctccagtgc caatgggatc
     1141 cattccagtt gttggaatgt ctccaaccct agtatcttct gttcccacag cagctgtgcc
     1201 ccccctggct aacggggctc cccctgttat acaacctctg cctgcatttg ctcatcctgc
     1261 agccacattg ccaaagagtt cttcctttag tagatctggt ccagggtcac aactaaacac
     1321 taaattacaa aaggcacagt catttgatgt ggccagtgtc ccaccagtgg cagagtgggc
     1381 tgttcctcag tcatcaagac tgaaatacag gcaattattc aatagtcatg acaaaactat
     1441 gagtggacac ttaacaggtc cccaagcaag aactattctt atgcagtcaa gtttaccaca
     1501 ggctcagctg gcttcaatat ggaatctttc tgacattgat caagatggaa aacttacagc
     1561 agaggaattt atcctggcaa tgcacctcat tgatgtagct atgtctggcc aaccactgcc
     1621 acctgtcctg cctccagaat acattccacc ttcttttaga agagttcgat ctggcagtgg
     1681 tatatctgtc ataagctcaa catctgtaga tcagaggcta ccagaggaac cagttttaga
     1741 agatgaacaa caacaattag aaaagaaatt acctgtaacg tttgaagata agaagcggga
     1801 gaactttgaa cgtggcaacc tggaactgga gaaacgaagg caagctctcc tggaacagca
     1861 gcgcaaggag caggagcgcc tggcccagct ggagcgggcg gagcaggaga ggaaggagcg
     1921 tgagcgccag gagcaagagc gcaaaagaca actggaactg gagaagcaac tggaaaagca
     1981 gcgggagcta gaacggcaga gagaggagga gaggaggaaa gaaattgaga ggcgagaggc
     2041 tgcaaaacgg gaacttgaaa ggcaacgaca acttgagtgg gaacggaatc gaaggcaaga
     2101 actactaaat caaagaaaca aagaacaaga ggacatagtt gtactgaaag caaagaaaaa
     2161 gactttggaa tttgaattag aagctctaaa tgataaaaag catcaactag aagggaaact
     2221 tcaagatatc agatgtcgat tgaccaccca aaggcaagaa attgagagca caaacaaatc
     2281 tagagagttg agaattgccg aaatcaccca tctacagcaa caattacagg aatctcagca
     2341 aatgcttgga agacttattc cagaaaaaca gatactcaat gaccaattaa aacaagttca
     2401 gcagaacagt ttgcacagag attcacttgt tacacttaaa agagccttag aagcaaaaga
     2461 actagctcgg cagcacctac gagaccaact ggatgaagtg gagaaagaaa ctagatcaaa
     2521 actacaggag attgatattt tcaataatca gctgaaggaa ctaagagaaa tacacaataa
     2581 gcaacaactc cagaagcaaa agtccatgga ggctgaacga ctgaaacaga aagaacaaga
     2641 acgaaagatc atagaattag aaaaacaaaa agaagaagcc caaagacgag ctcaggaaag
     2701 ggacaagcag tggctggagc atgtgcagca ggaggacgag catcagagac caagaaaact
     2761 ccacgaagag gaaaaactga aaagggagga gagtgtcaaa aagaaggatg gcgaggaaaa
     2821 aggcaaacag gaagcacaag acaagctggg tcggcttttc catcaacacc aagaaccagc
     2881 taagccagct gtccaggcac cctggtccac tgcagaaaaa ggtccactta ccatttctgc
     2941 acaggaaaat gtaaaagtgg tgtattaccg ggcactgtac ccctttgaat ccagaagcca
     3001 tgatgaaatc actatccagc caggagacat agtcatggtg gatgaaagcc aaactggaga
     3061 acccggctgg cttggaggag aattaaaagg aaagacaggg tggttccctg caaactatgc
     3121 agagaaaatc ccagaaaatg aggttcccgc tccagtgaaa ccagtgactg attcaacatc
     3181 tgcccctgcc cccaaactgg ccttgcgtga gacccccgcc cctttggcag taacctcttc
     3241 agagccctcc acgaccccta ataactgggc cgacttcagc tccacgtggc ccaccagcac
     3301 gaatgagaaa ccagaaacgg ataactggga tgcatgggca gcccagccct ctctcaccgt
     3361 tccaagtgcc ggccagttaa ggcagaggtc cgcctttact ccagccacgg ccactggctc
     3421 ctccccgtct cctgtgctag gccagggtga aaaggtggag gggctacaag ctcaagccct
     3481 atatccttgg agagccaaaa aagacaacca cttaaatttt aacaaaaatg atgtcatcac
     3541 cgtcctggaa cagcaagaca tgtggtggtt tggagaagtt caaggtcaga agggttggtt
     3601 ccccaagtct tacgtgaaac tcatttcagg gcccataagg aagtctacaa gcatggattc
     3661 tggttcttca gagagtcctg ctagtctaaa gcgagtagcc tctccagcag ccaagccggt
     3721 cgtttcggga gaagaattta ttgccatgta cacttacgag agttctgagc aaggagattt
     3781 aacctttcag caaggggatg tgattttggt taccaagaaa gatggtgact ggtggacagg
     3841 aacagtgggc gacaaggccg gagtcttccc ttctaactat gtgaggctta aagattcaga
     3901 gggctctgga actgctggga aaacagggag tttaggaaaa aaacctgaaa ttgcccaggt
     3961 tattgcctca tacaccgcca ccggccccga gcagctcact ctcgcccctg gtcagctgat
     4021 tttgatccga aaaaagaacc caggtggatg gtgggaagga gagctgcaag cacgtgggaa
     4081 aaagcgccag ataggctggt tcccagctaa ttatgtaaag cttctaagcc ctgggacgag
     4141 caaaatcact ccaacagagc cacctaagtc aacagcatta gcggcagtgt gccaggtaag
     4201 tatcaaggtt acaagacagg tttaaggaga ccaataagct attattcgta taggatactt
     4261 tatacgaagt tattaagtta tggagaccgg tgtggcacgt gcggaccgag cggccgcagg
     4321 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     4381 ggcgaccaaa ggtcgcccga cgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     4441 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4501 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4561 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     4621 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4681 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     4741 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4801 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     4861 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     4921 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     4981 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     5041 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     5101 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5161 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5221 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5281 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5341 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5401 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5461 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5521 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5581 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5641 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5701 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5761 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5821 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5881 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5941 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6001 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6061 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6121 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6181 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6241 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6301 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6361 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6421 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6481 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6541 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6601 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6661 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6721 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6781 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6841 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6901 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6961 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7021 tttgctggcc ttttgctcac atgt
//
