LOCUS       sequence-465825-        6418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' AAVLINK plasmid for PXDN expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6418)
  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  JOURNAL   Unpublished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 6418)
  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            "Unpublished"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' PXDN
FEATURES             Location/Qualifiers
     source          1..6418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..397
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             410..678
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             715..741
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     repeat_region   3681..3821
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3681..3810
                     /label=AAV2 ITR
     rep_origin      3896..4351
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3983..4002)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4193..4214
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4368..4387)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4487..4509
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4547..4565)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4633..4737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4738..4806,4807..5598)
                     /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(4956..4975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5769..6357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6258..6277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtggatc tgcgatcgct ccggtgcccg
      181 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
      241 ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
      301 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
      361 cgttcttttt cgcaacgggt ttgccgccag aacacagctg aagcttcgag gggctcgcat
      421 ctctccttca cgcgcccgcc gccctacctg aggccgccat ccacgccggt tgagtcgcgt
      481 tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa
      541 gctcaggtcg agaccgggcc tttgtccggc gctcccttgg agcctaccta gactcagccg
      601 gctctccacg ctttgcctga ccctgcttgc tcaactctac gtctttgttt cgttttctgt
      661 tctgcgccgt tacagatcca agctgtgacc ggcgcctacc gtacggccac catgtaccct
      721 tatgacgtac ctgactatgc tggatctgcc aagcgctcca ggggccccgg gcgccgctgc
      781 ctgttggcgc tcgtgctgtt ctgcgcctgg gggacgctgg ccgtggtggc ccagaagccg
      841 ggcgcagggt gtccgagccg ctgcctgtgc ttccgcacca ccgtgcgctg catgcatctg
      901 ctgctggagg ccgtgcccgc cgtggcgccg cagacctcca tcctagatct tcgctttaac
      961 agaatcagag agatccaacc tggggcattc aggcggctga ggaacttgaa cacattgctt
     1021 ctcaataata atcagatcaa gaggatacct agtggagcat ttgaagactt ggaaaattta
     1081 aaatatctct atctgtacaa gaatgagatc cagtcaattg acaggcaagc atttaaggga
     1141 cttgcctctc tagagcaact atacctgcac tttaatcaga tagaaacttt ggacccagat
     1201 tcgttccagc atctcccgaa gctcgagagg ctatttttgc ataacaaccg gattacacat
     1261 ttagttccag ggacatttaa tcacttggaa tctatgaaga gattgcgact ggactcaaac
     1321 acacttcact gcgactgtga aatcctgtgg ttggcggatt tgctgaaaac ctacgcggag
     1381 tcggggaacg cgcaggcagc ggccatctgt gaatatccca gacgcatcca gggacgctca
     1441 gtggcaacca tcaccccgga agagctgaac tgtgaaaggc cccggatcac ctccgagccc
     1501 caggacgcag atgtgacctc ggggaacacc gtgtacttca cctgcagagc cgaaggcaac
     1561 cccaagcctg agatcatctg gctgcgaaac aataatgagc tgagcatgaa gacagattcc
     1621 cgcctaaact tgctggacga tgggaccctg atgatccaga acacacagga gacagaccag
     1681 ggtatctacc agtgcatggc aaagaacgtg gccggagagg tgaagacgca agaggtgacc
     1741 ctcaggtact tcgggtctcc agctcgaccc acttttgtaa tccagccaca gaatacagag
     1801 gtgctggttg gggagagcgt cacgctggag tgcagcgcca caggccaccc cccgccgcgg
     1861 atctcctgga cgagaggtga ccgcacaccc ttgccagttg acccgcgggt gaacatcacg
     1921 ccttctggcg ggctttacat acagaacgtc gtacaggggg acagcggaga gtatgcgtgc
     1981 tctgcgacca acaacattga cagcgtccat gccaccgctt tcatcatcgt ccaggctctt
     2041 cctcagttca ctgtgacgcc tcaggacaga gtcgttattg agggccagac cgtggatttc
     2101 cagtgtgaag ccaagggcaa cccgccgccc gtcatcgcct ggaccaaggg agggagccag
     2161 ctctccgtgg accggcggca cctggtcctg tcatcgggaa cacttagaat ctctggtgtt
     2221 gccctccacg accagggcca gtacgaatgc caggctgtca acatcatcgg ctcccagaag
     2281 gtcgtggccc acctgactgt gcagcccaga gtcaccccag tgtttgccag cattcccagc
     2341 gacacaacag tggaggtggg cgccaatgtg cagctcccgt gcagctccca gggcgagccc
     2401 gagccagcca tcacctggaa caaggatggg gttcaggtga cagaaagtgg aaaatttcac
     2461 atcagccctg aaggattctt gaccatcaat gacgttggcc ctgcagacgc aggtcgctat
     2521 gagtgtgtgg cccggaacac cattgggtcg gcctcggtga gcatggtgct cagtgtgaac
     2581 gttcctgacg tcagtcgaaa tggagatccg tttgtagcta cctccatcgt ggaagcgatt
     2641 gcgactgttg acagagctat aaactcaacc cgaacacatt tgtttgacag ccgtcctcgt
     2701 tctccaaatg atttgctggc cttgttccgg tatccgaggg atccttacac agttgaacag
     2761 gcacgggcgg gagaaatctt tgaacggaca ttgcagctca ttcaggagca tgtacagcat
     2821 ggcttgatgg tcgacctcaa cggaacaagt taccactaca acgacctggt gtctccacag
     2881 tacctgaacc tcatcgcaaa cctgtcgggc tgtaccgccc accggcgcgt gaacaactgc
     2941 tcggacatgt gcttccacca gaagtaccgg acgcacgacg gcacctgtaa caacctgcag
     3001 caccccatgt ggggcgcctc gctgaccgcc ttcgagcgcc tgctgaaatc cgtgtacgag
     3061 aatggcttca acacccctcg gggcatcaac ccccaccgac tgtacaacgg gcacgccctt
     3121 cccatgccgc gcctggtgtc caccaccctg atcgggacgg agaccgtcac acccgacgag
     3181 cagttcaccc acatgctgat gcagtggggc cagttcctgg accacgacct cgactccacg
     3241 gtggtggccc tgagccaggc acgcttctcc gacggacagc actgcagcaa cgtgtgcagc
     3301 aacgaccccc cctgcttctc tgtcatgatc ccccccaatg actcccgggc caggagcggg
     3361 gcccgctgca tgttcttcgt gcgctccagc cctgtgtgcg gcagcggcat gacttcgctg
     3421 ctcatgaact ccgtgtaccc gcgggagcag atcaaccagc tcacctccta catcgacgca
     3481 tccaacgtgt acgggagcac ggagcatgag gcccgcagca tccgcgacct ggccagccac
     3541 cgcggcctgc tgcggcaggt aagtatcaag gttacaagac aggtttaagg agaccaataa
     3601 gctattattc gtataggata ctttatacga agttattaag ttatggagac cggtgtggca
     3661 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     3721 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3781 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     3841 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     3901 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3961 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4021 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4081 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     4141 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4201 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     4261 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4321 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4381 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4441 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4501 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4561 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4621 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4681 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4741 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4801 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4861 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4921 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4981 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5041 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5101 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5161 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5221 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5281 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5341 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5401 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5461 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5521 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5581 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5641 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5701 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5761 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5821 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5881 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5941 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6001 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6061 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6121 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6181 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6241 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6301 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6361 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
