LOCUS       sequence-479616-        6195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for NINL expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6195)
  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  JOURNAL   Unpublished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 6195)
  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            "Unpublished"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' NINL
FEATURES             Location/Qualifiers
     source          1..6195
                     /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   3458..3598
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3458..3587
                     /label=AAV2 ITR
     rep_origin      3673..4128
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3760..3779)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3970..3991
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4145..4164)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4264..4286
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4324..4342)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4410..4514
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4515..4583,4584..5375)
                     /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(4733..4752)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5546..6134
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6035..6054
                     /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 ctggatctga tgaagaagag aaccactatg tctcgcagct
      781 cagggaagtc tacagcagct gcgacaccac ggggactggc tttctggacc gccaggagct
      841 gacccagctc tgccttaagc ttcacctgga gcagcagctg cccgtcctcc tgcagacgct
      901 tctcggaaac gaccatttcg ccagggttaa ctttgaggaa tttaaggaag gttttgtggc
      961 tgtgttgtct tcaaatgctg gtgttcgccc ctcagatgaa gacagtagtt ctttggaatc
     1021 agctgcctcc agtgccatcc ctccaaagta tgtgaatggt tctaagtggt atggccgtcg
     1081 gagccggcct gagctatgtg acgctgccac agaagccaga cgcgtgccgg agcagcaaac
     1141 ccaggccagc ctgaaaagtc acctctggcg ctcagcgtct ctggagagcg tggagagtcc
     1201 caagtcagat gaagaggccg agagcactaa agaagctcag aatgaattat ttgaagcaca
     1261 aggacagctg cagacctggg attctgagga ctttgggagc ccccagaagt cctgcagccc
     1321 ctcctttgac accccagaga gccagatccg gggcgtgtgg gaagagctgg gggtgggcag
     1381 cagcggacac ctgagcgagc aggagctggc tgtggtctgc cagagcgtcg ggctccaggg
     1441 actcgagaaa gaggaactcg aagacctgtt taacaaactg gatcaagacg gagacggcaa
     1501 agtgagtctt gaggaattcc agcttggcct cttcagtcat gagcccgcgc tacttctaga
     1561 gtcttccact cgggttaaac cgagcaaggc ttggtctcat taccaggtcc cagaggagag
     1621 cggctgccac accaccacaa cctcatccct cgtgtccctg tgctccagcc tgcgcctctt
     1681 ctccagcatt gacgatggtt ctggcttcgc ttttcctgat caggtcctgg ccatgtggac
     1741 ccaggagggg attcagaatg gcagggagat cttgcagagc ctggacttca gcgtggacga
     1801 gaaggtgaac cttctggagc tgacctgggc ccttgacaac gagctcatga cagtggacag
     1861 tgccgtccag caggcagccc tggcctgcta ccaccaggag ctgagctacc agcaagggca
     1921 ggtggagcag ctggcaaggg agcgtgacaa ggcaaggcag gacctggaga gggccgagaa
     1981 gaggaacctg gagtttgtga aagagatgga cgactgccac tccaccctgg agcagctcac
     2041 ggagaagaaa atcaagcatc tggagcaggg gtaccgggaa aggctgagcc tcctgcggtc
     2101 tgaggtggag gcggagcgag agctgttctg ggagcaggcc cacaggcaga gggccgcgct
     2161 ggagtgggac gtggggcgcc tgcaggctga ggaggctggc ctccgcgaga agctgaccct
     2221 ggccctgaag gaaaacagtc gcctacagaa ggagattgtg gaagtggtgg aaaagctttc
     2281 ggattcggag aggctggccc tgaagctgca gaaggacctg gagtttgtgc tgaaggacaa
     2341 gctggagcca cagagtgcag agctcctggc ccaggaggag cggttcgcag cagtcctgaa
     2401 ggaatacgag ctcaagtgcc gggacctgca ggaccgcaac gatgagctgc aagctgagct
     2461 ggaaggcctg tgggcgcggc tgcccaagaa ccggcacagc ccctcatgga gcccggatgg
     2521 gcgcagacgg cagctccctg gactcggccc agcaggcatt tcattcctgg gtaattctgc
     2581 tccagtgagt atagaaacgg agctgatgat ggagcaggta aaggagcatt accaagacct
     2641 caggacccag ctggagacca aggtaaatta ctacgaaagg gaaattgcgg cactgaaaag
     2701 gaactttgag aaggagagga aggacatgga gcaggctcgc cggcgcgagg tcagcgtgct
     2761 ggagggtcag aaggccgacc tggaggagct ccacgagaag tctcaggagg tcatctgggg
     2821 cctgcaggag cagctgcagg acacagcccg cggccccgag cctgagcaga tgggcctggc
     2881 accctgctgc acccaggcac tgtgtggcct ggccctgcgg catcacagcc acctgcagca
     2941 gatcaggaga gaggctgagg cggagctgag tggagagctg tcggggctgg gagccctgcc
     3001 cgctcgcaga gacctgacct tggagctgga ggagccgccg cagggacccc tgccacgcgg
     3061 gagccagagg tcggagcagc tggagctgga gagggcactg aagctgcagc cctgtgcgag
     3121 cgagaagcgc gcccagatgt gcgtatcgtt ggccctcgag gaggaggagt tggagcttgc
     3181 ccgcgggaag cgagtggacg ggccctccct ggaagcagag atgcaggccc tgccgaaaga
     3241 tgggctggtg gcaggaagtg gccaggaggg cacacgtggc ctcctaccac tgcgtccggg
     3301 ctgtggggag cggccactgg cctggctggc cccaggtaag tatcaaggtt acaagacagg
     3361 tttaaggaga ccaataagct attattcgta taggatactt tatacgaagt tattaagtta
     3421 tggagaccgg tgtggcacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     3481 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3541 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     3601 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     3661 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3721 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     3781 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3841 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     3901 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3961 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     4021 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     4081 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     4141 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4201 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4261 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     4321 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4381 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4441 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4501 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4561 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4621 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4681 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4741 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4801 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4861 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4921 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4981 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5041 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5101 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5161 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5221 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5281 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5341 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5401 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5461 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5521 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5581 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5641 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5701 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5761 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5821 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5881 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5941 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6001 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6061 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6121 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6181 cttttgctca catgt
//
