LOCUS       sequence-476893-        6310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for NEXMIF expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6310)
  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  JOURNAL   Unpublished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 6310)
  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            "Unpublished"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' NEXMIF
FEATURES             Location/Qualifiers
     source          1..6310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1088..1228
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1088..1217
                     /label=AAV2 ITR
     rep_origin      1303..1758
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1390..1409)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1600..1621
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1775..1794)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1894..1916
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1954..1972)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2040..2144
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2145..2213,2214..3005)
                     /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(2363..2382)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3176..3764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3665..3684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3826..3955
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        4000..4211
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             4224..4492
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4529..4555
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 aaaggcttct ggcaaaagaa gaagaagcag agaaacacca acacggactc catcaagaca
       61 cctttttccc aaaagcaaag ctttgaacca ggtagctttg aggtgtcatt tctgccacct
      121 gctcgcaaac gaaaatctaa acttggcaac aggcacagga ttcaaagaat cccatccatt
      181 gaaatttcag caagtagtaa acagatttca ttatgtaatg atcagaggca cgctagtaat
      241 cataaagaag atggaggcct aaaaggtaca ctgaagtcag cacctctggg tgctcctagc
      301 tgtgcaaatg gatcacattt aaatgacatc acaggccctg actcagtgaa agtcaaagcc
      361 caagacacag agtttaaggg gccagagagg aaagtgctca ataaaatcaa atttaaaagt
      421 gaagctaggt taaaatctaa gaaagtcaaa gctgctgggc aagaaagcaa gccaattgtt
      481 caaatgagcc ctctcttgga aaaccaatcc tcaaaggcta atttaaagaa tgaagttatt
      541 cctgggacat caaacagttc ccgtctatct gaatttcatg aggcaaaggc tgctaagagt
      601 tccacttttc taccaacgac atgctcttct gaaatgcctt tatcatctgc taatgttacc
      661 actaatatac ctgttatccc gggagggtat ctgcagacat tgttagatgc ttctgacttg
      721 tcaaataaca ctagtatctc atacttcagc caccattctc cagagcaaaa tgaaggcagc
      781 ctcactcaaa ccgaaaaatc atttgtaccc ctccagccta cccaggactg tgtgctcacc
      841 tcatcctctg actctgagct gcagcagtca tctcataact tcaaaatgga atcaagcaac
      901 tatagaaatg tgtggcccaa caaggctact tctggcaccc aggaattcat ggctgaagtc
      961 tcaaggtaag tatcaaggtt acaagacagg tttaaggaga ccaataagct attattcgta
     1021 taggatactt tatacgaagt tattaagtta tggagaccgg tgtggcacgt gcggaccgag
     1081 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     1141 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     1201 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     1261 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     1321 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     1381 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     1441 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     1501 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     1561 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     1621 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     1681 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     1741 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     1801 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     1861 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     1921 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     1981 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     2041 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2101 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2161 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2221 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2281 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2341 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2401 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2461 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2521 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2581 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2641 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2701 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2761 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2821 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2881 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2941 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3001 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3061 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3121 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3301 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3661 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3721 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3781 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc
     3841 gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca
     3901 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc
     3961 cgcacgcgtg gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca
     4021 cagtccccga gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc
     4081 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     4141 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     4201 ccagaacaca gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta
     4261 cctgaggccg ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct
     4321 cctgaactgc gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc
     4381 cggcgctccc ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc
     4441 ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt
     4501 gaccggcgcc taccgtacgg ccaccatgta cccttatgac gtacctgact atgctggatc
     4561 tgataaccaa caagataagg ctattgttgc ctcagccaac ggagaaaaca ctctgattaa
     4621 tggggtcaaa gaaaatgact cagaggacca ggatgtggca atgaagtcat ttgcagctct
     4681 agaagctgct gcacctatcc agcctacacc ggtggcacaa aaagagaccc tgatgtatcc
     4741 cagaggtctc ctgcctctac cctctaagaa gccctgtatg cagagcccgc cctctccttt
     4801 gggcctgatt gaagcacccg aacatgctgc taatagtgct tctgtgaatg ccatctccct
     4861 cacatctggc attgcaaaag gcctgaacac atggtcactt cccaatgaat gtgagaaagc
     4921 tccatttgcc ataatggagc ctgcaggcat gtcagctctg aatggggact gtctcatgca
     4981 gccaagtcgg acttgcttag gctgcttcat ggaatccaag gatgcagtag atcctgagcc
     5041 agggatcagt ctgaaagttg gtgatctaaa tagggattat gaaacgtgtg cagtctctga
     5101 tatagggatt cagtgtatta atgctggaga aaatatgaaa tatggagagc agctgctctc
     5161 agaccagctc ctaggcttcc ccctgcataa gtcaagggca ggagacagac gagaaactga
     5221 gaaacctgac attgacttgg aggatccggc tcagaaaagc tattatgagg cattactgtt
     5281 agacaagtgc aatacagaag aagctttgct tgcaaattcc aatcaggatt ggggttactt
     5341 cgagactttt attagtgaaa gtaagattga actgcttgac ctctgttcca aaaatgagct
     5401 gtctgtcaac ctattctctg aagaagatgt ggataactac atgtttgatg atgatgaatc
     5461 aacactaggc agtgatgtct gctccctgaa aattcgatat gaatcctttc aggacaatgt
     5521 tcgagacaag actactcttt tgatgcagga agatgcccaa ttcaactttt ttcccagcgt
     5581 ctttactacc tgccccaagc gagagtctaa gagtggggcc ctgaagcaga gcagtgattt
     5641 ttcccaattc aaggtccctg atgtgagcat catctggggg gaggaagata aaaacttgga
     5701 caagaagaaa ggcaaagagg aaggacagga agacaaaggt gtagagaaga aagatggaaa
     5761 ggataatgga gaaaagcctg ccttaaataa accatgcagt gggactgaag tagagcaact
     5821 taagaatcca aagcagggcc atcttgctaa ttccctggag acatcaggga gtttcagtga
     5881 tgatagttcc ttcattgaga tctcatatga tgctatgggt gagatcaagg actgtagtcg
     5941 ctatatggct cgggacacta attctggcag ctcctcctcc caacagaact atgggctgcg
     6001 agccaagaga aaagtcagat acagtgaaga ttatctatat gatgttgact cactagaggg
     6061 tgaaaaagta aatgagagga aggaatggct gccagttggt tccaaagagg aagatgatga
     6121 tgaatggtgt cccaaaaaga gaagaaaagt aacccgtaag gagccccctg ttattatcaa
     6181 atatatcatc attaatcgct ttaaaggtga gaagaacatg ctggtgaagt tgggtaaggt
     6241 ggatgccagt gagacaacag tgaatttgag tgagaatcag ctcaacaaat atgccaagct
     6301 ggcacccttg
//
