LOCUS       sequence-476888-        5366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for UBN2 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5366)
  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  JOURNAL   Unpublished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 5366)
  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            "Unpublished"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' UBN2
FEATURES             Location/Qualifiers
     source          1..5366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             31..57
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     repeat_region   1956..2096
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1956..2085
                     /label=AAV2 ITR
     rep_origin      2171..2626
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2258..2277)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2468..2489
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2643..2662)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2762..2784
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2822..2840)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2908..3012
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3013..3081,3082..3873)
                     /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(3231..3250)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4044..4632
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4533..4552
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4694..4823
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        4868..5079
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5092..5360
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
ORIGIN
        1 gtgaccggcg cctaccgtac ggccaccatg tacccttatg acgtacctga ctatgctgga
       61 tctgcggagc cgcgcagagt agcgttcatt agcttgtcac cggtgcggcg gcgcgaggcc
      121 gagtacccgg ggcccgagcg tgagcccgag tacccccgcg agcccccccg gctggtgcgc
      181 ctggagctgg tgcttaagga ccccaccgac gagagctgcg tggagttcag ttacccggag
      241 ctgctgctgt gcggagaaca acggaagaag ctcattcaca cagaagaccc atttaatgat
      301 gaacatcagg agaggcaaga ggtggaaatg ttggctaaga agtttgaaat gaaatatggt
      361 gggaaacccc gtaaacaccg gaaggatcgg ctacaagatt taattgatat aggctttggc
      421 tatgatgaga cagatccatt tattgataac tcagaggctt atgatgaatt agttcccgct
      481 tctctaacaa caaaatatgg aggcttttat atcaacactg gcactctaca gtttcgccaa
      541 gcttcagata ctgaagaaga tgatattaca gacaaccaaa agcacaagcc acccaaggtc
      601 cccaaaataa aagaagatga tattgagatg aagaagcgga agcggaaaga ggaaggggaa
      661 aaggagaaga agccaaggaa aaaagttccc aaacaactgg gagttgtggc tctaaattca
      721 cacaagtctg aaaaaaagaa gaaacgttat aaagattctc tttctctagc tgccatgatt
      781 agaaaattcc agaaagagaa ggatgcatta aagaaggagt ctaaccccaa agtcccagtg
      841 accttgtcaa ccccttctct gaataaaccc ccatgtgctg ctgcagcact ggggaatgac
      901 gtcccggact taaatctgag cagcggtgat ccagaccttc ccatttttgt tagcacaaat
      961 gaacatgagc tgtttcagga agctgaaaat gccctagaga tgctagatga ttttgacttc
     1021 gacagattac tggatgctgc ttctgatggt agccccctat ctgagtcggg gggtgaaaat
     1081 ggaaccacca cccagccaac ctacacttct caggttatgc ccaaagtggt acctacactc
     1141 ccagagggtc tacctgtact tcttgaaaaa cgtatcgaag accttcgtgt agctgccaaa
     1201 ctttttgatg aagaaggaag gaaaaaattc tttacacagg atatgaataa tattcttctg
     1261 gacattgagt tacagctaca agaactaggc cctgtcattc gcagtggtgt ctactcccac
     1321 cttgaagctt ttgtgccatg caataaagaa acactagtaa aacgtctgaa gaagttacat
     1381 ctcaatgtcc aggatgatcg tttaagagaa cctctgcaaa aactgaaact ggctgttagc
     1441 aatgtcatgc ctgaacagct atttaaatac caggaggact gccaggctcg tagtcaagct
     1501 aagtgtgcca aattgcagac agatgaagaa cgagaaaaaa atggatctga agaggatgat
     1561 gatgagaaac caggaaaacg tgtcatagga ccaagaaaga aattccactg ggatgacact
     1621 atcagaactt tgttatgtaa ccttgttgag atcaaattgg gatgctatga gttagaacca
     1681 aataaaagcc agtctgctga agattatctt aagtctttta tggagacaga agtgaagccc
     1741 ctgtggccta agggctggat gcaggcaaga atgcttttta aggaaagccg gagtgttcat
     1801 aatcatctta cttctgctcc ggcaaagaaa aaggtaagta tcaaggttac aagacaggtt
     1861 taaggagacc aataagctat tattcgtata ggatacttta tacgaagtta ttaagttatg
     1921 gagaccggtg tggcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     1981 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2041 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     2101 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     2161 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2221 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     2281 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2341 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     2401 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2461 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     2521 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     2581 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     2641 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     2701 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     2761 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     2821 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2881 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2941 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3001 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3061 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3121 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3181 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3241 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3301 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3361 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3421 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3481 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3541 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3601 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3661 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3721 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3781 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3841 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3901 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3961 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4021 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4081 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4141 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4201 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4261 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4321 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4381 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4441 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4501 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4561 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4621 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4681 tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg
     4741 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
     4801 caactccatc actaggggtt cctgcggccg cacgcgtgga tctgcgatcg ctccggtgcc
     4861 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     4921 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     4981 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     5041 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc
     5101 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc
     5161 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta
     5221 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc
     5281 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct
     5341 gttctgcgcc gttacagatc caagct
//
