LOCUS       sequence-491862-        6374 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 6374)
  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  JOURNAL   Unpublished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 6374)
  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            "Unpublished"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..6374
                     /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"
     regulatory      695..704
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3637..3777
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3637..3766
                     /label=AAV2 ITR
     rep_origin      3852..4307
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3939..3958)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4149..4170
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4324..4343)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4443..4465
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4503..4521)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4589..4693
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4694..4762,4763..5554)
                     /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(4912..4931)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5725..6313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6214..6233
                     /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 atggccaagc gctccagggg
      721 ccccgggcgc cgctgcctgt tggcgctcgt gctgttctgc gcctggggga cgctggccgt
      781 ggtggcccag aagccgggcg cagggtgtcc gagccgctgc ctgtgcttcc gcaccaccgt
      841 gcgctgcatg catctgctgc tggaggccgt gcccgccgtg gcgccgcaga cctccatcct
      901 agatcttcgc tttaacagaa tcagagagat ccaacctggg gcattcaggc ggctgaggaa
      961 cttgaacaca ttgcttctca ataataatca gatcaagagg atacctagtg gagcatttga
     1021 agacttggaa aatttaaaat atctctatct gtacaagaat gagatccagt caattgacag
     1081 gcaagcattt aagggacttg cctctctaga gcaactatac ctgcacttta atcagataga
     1141 aactttggac ccagattcgt tccagcatct cccgaagctc gagaggctat ttttgcataa
     1201 caaccggatt acacatttag ttccagggac atttaatcac ttggaatcta tgaagagatt
     1261 gcgactggac tcaaacacac ttcactgcga ctgtgaaatc ctgtggttgg cggatttgct
     1321 gaaaacctac gcggagtcgg ggaacgcgca ggcagcggcc atctgtgaat atcccagacg
     1381 catccaggga cgctcagtgg caaccatcac cccggaagag ctgaactgtg aaaggccccg
     1441 gatcacctcc gagccccagg acgcagatgt gacctcgggg aacaccgtgt acttcacctg
     1501 cagagccgaa ggcaacccca agcctgagat catctggctg cgaaacaata atgagctgag
     1561 catgaagaca gattcccgcc taaacttgct ggacgatggg accctgatga tccagaacac
     1621 acaggagaca gaccagggta tctaccagtg catggcaaag aacgtggccg gagaggtgaa
     1681 gacgcaagag gtgaccctca ggtacttcgg gtctccagct cgacccactt ttgtaatcca
     1741 gccacagaat acagaggtgc tggttgggga gagcgtcacg ctggagtgca gcgccacagg
     1801 ccaccccccg ccgcggatct cctggacgag aggtgaccgc acacccttgc cagttgaccc
     1861 gcgggtgaac atcacgcctt ctggcgggct ttacatacag aacgtcgtac agggggacag
     1921 cggagagtat gcgtgctctg cgaccaacaa cattgacagc gtccatgcca ccgctttcat
     1981 catcgtccag gctcttcctc agttcactgt gacgcctcag gacagagtcg ttattgaggg
     2041 ccagaccgtg gatttccagt gtgaagccaa gggcaacccg ccgcccgtca tcgcctggac
     2101 caagggaggg agccagctct ccgtggaccg gcggcacctg gtcctgtcat cgggaacact
     2161 tagaatctct ggtgttgccc tccacgacca gggccagtac gaatgccagg ctgtcaacat
     2221 catcggctcc cagaaggtcg tggcccacct gactgtgcag cccagagtca ccccagtgtt
     2281 tgccagcatt cccagcgaca caacagtgga ggtgggcgcc aatgtgcagc tcccgtgcag
     2341 ctcccagggc gagcccgagc cagccatcac ctggaacaag gatggggttc aggtgacaga
     2401 aagtggaaaa tttcacatca gccctgaagg attcttgacc atcaatgacg ttggccctgc
     2461 agacgcaggt cgctatgagt gtgtggcccg gaacaccatt gggtcggcct cggtgagcat
     2521 ggtgctcagt gtgaacgttc ctgacgtcag tcgaaatgga gatccgtttg tagctacctc
     2581 catcgtggaa gcgattgcga ctgttgacag agctataaac tcaacccgaa cacatttgtt
     2641 tgacagccgt cctcgttctc caaatgattt gctggccttg ttccggtatc cgagggatcc
     2701 ttacacagtt gaacaggcac gggcgggaga aatctttgaa cggacattgc agctcattca
     2761 ggagcatgta cagcatggct tgatggtcga cctcaacgga acaagttacc actacaacga
     2821 cctggtgtct ccacagtacc tgaacctcat cgcaaacctg tcgggctgta ccgcccaccg
     2881 gcgcgtgaac aactgctcgg acatgtgctt ccaccagaag taccggacgc acgacggcac
     2941 ctgtaacaac ctgcagcacc ccatgtgggg cgcctcgctg accgccttcg agcgcctgct
     3001 gaaatccgtg tacgagaatg gcttcaacac ccctcggggc atcaaccccc accgactgta
     3061 caacgggcac gcccttccca tgccgcgcct ggtgtccacc accctgatcg ggacggagac
     3121 cgtcacaccc gacgagcagt tcacccacat gctgatgcag tggggccagt tcctggacca
     3181 cgacctcgac tccacggtgg tggccctgag ccaggcacgc ttctccgacg gacagcactg
     3241 cagcaacgtg tgcagcaacg accccccctg cttctctgtc atgatccccc ccaatgactc
     3301 ccgggccagg agcggggccc gctgcatgtt cttcgtgcgc tccagccctg tgtgcggcag
     3361 cggcatgact tcgctgctca tgaactccgt gtacccgcgg gagcagatca accagctcac
     3421 ctcctacatc gacgcatcca acgtgtacgg gagcacggag catgaggccc gcagcatccg
     3481 cgacctggcc agccaccgcg gcctgctgcg gcaggtaagt atcaaggtta caagacaggt
     3541 ttaaggagac caataagcta ttattcgtat aggatacttt atacgaagtt attaagttat
     3601 ggagaccggt gtggcacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     3661 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3721 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     3781 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     3841 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3901 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     3961 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4021 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4081 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4141 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     4201 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     4261 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     4321 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4381 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4441 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     4501 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4561 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4621 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4681 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4741 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4801 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4861 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4921 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4981 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5041 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5101 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5161 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5221 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5281 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5341 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5401 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5461 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5521 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5581 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5641 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5701 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5761 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5821 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5881 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5941 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6001 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6061 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6121 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6181 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6241 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6301 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6361 ttttgctcac atgt
//
