LOCUS       sequence-476884-        6407 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 6407)
  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  JOURNAL   Unpublished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 6407)
  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            "Unpublished"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..6407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2823..2963
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2823..2952
                     /label=AAV2 ITR
     rep_origin      3038..3493
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3125..3144)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3335..3356
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3510..3529)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3629..3651
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3689..3707)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3775..3879
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3880..3948,3949..4740)
                     /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(4098..4117)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4911..5499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5400..5419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5561..5690
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        5735..5946
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5959..6227
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             6264..6290
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 cgctgcctgt gcttccgcac caccgtgcgc tgcatgcatc tgctgctgga ggccgtgccc
       61 gccgtggcgc cgcagacctc catcctagat cttcgcttta acagaatcag agagatccaa
      121 cctggggcat tcaggcggct gaggaacttg aacacattgc ttctcaataa taatcagatc
      181 aagaggatac ctagtggagc atttgaagac ttggaaaatt taaaatatct ctatctgtac
      241 aagaatgaga tccagtcaat tgacaggcaa gcatttaagg gacttgcctc tctagagcaa
      301 ctatacctgc actttaatca gatagaaact ttggacccag attcgttcca gcatctcccg
      361 aagctcgaga ggctattttt gcataacaac cggattacac atttagttcc agggacattt
      421 aatcacttgg aatctatgaa gagattgcga ctggactcaa acacacttca ctgcgactgt
      481 gaaatcctgt ggttggcgga tttgctgaaa acctacgcgg agtcggggaa cgcgcaggca
      541 gcggccatct gtgaatatcc cagacgcatc cagggacgct cagtggcaac catcaccccg
      601 gaagagctga actgtgaaag gccccggatc acctccgagc cccaggacgc agatgtgacc
      661 tcggggaaca ccgtgtactt cacctgcaga gccgaaggca accccaagcc tgagatcatc
      721 tggctgcgaa acaataatga gctgagcatg aagacagatt cccgcctaaa cttgctggac
      781 gatgggaccc tgatgatcca gaacacacag gagacagacc agggtatcta ccagtgcatg
      841 gcaaagaacg tggccggaga ggtgaagacg caagaggtga ccctcaggta cttcgggtct
      901 ccagctcgac ccacttttgt aatccagcca cagaatacag aggtgctggt tggggagagc
      961 gtcacgctgg agtgcagcgc cacaggccac cccccgccgc ggatctcctg gacgagaggt
     1021 gaccgcacac ccttgccagt tgacccgcgg gtgaacatca cgccttctgg cgggctttac
     1081 atacagaacg tcgtacaggg ggacagcgga gagtatgcgt gctctgcgac caacaacatt
     1141 gacagcgtcc atgccaccgc tttcatcatc gtccaggctc ttcctcagtt cactgtgacg
     1201 cctcaggaca gagtcgttat tgagggccag accgtggatt tccagtgtga agccaagggc
     1261 aacccgccgc ccgtcatcgc ctggaccaag ggagggagcc agctctccgt ggaccggcgg
     1321 cacctggtcc tgtcatcggg aacacttaga atctctggtg ttgccctcca cgaccagggc
     1381 cagtacgaat gccaggctgt caacatcatc ggctcccaga aggtcgtggc ccacctgact
     1441 gtgcagccca gagtcacccc agtgtttgcc agcattccca gcgacacaac agtggaggtg
     1501 ggcgccaatg tgcagctccc gtgcagctcc cagggcgagc ccgagccagc catcacctgg
     1561 aacaaggatg gggttcaggt gacagaaagt ggaaaatttc acatcagccc tgaaggattc
     1621 ttgaccatca atgacgttgg ccctgcagac gcaggtcgct atgagtgtgt ggcccggaac
     1681 accattgggt cggcctcggt gagcatggtg ctcagtgtga acgttcctga cgtcagtcga
     1741 aatggagatc cgtttgtagc tacctccatc gtggaagcga ttgcgactgt tgacagagct
     1801 ataaactcaa cccgaacaca tttgtttgac agccgtcctc gttctccaaa tgatttgctg
     1861 gccttgttcc ggtatccgag ggatccttac acagttgaac aggcacgggc gggagaaatc
     1921 tttgaacgga cattgcagct cattcaggag catgtacagc atggcttgat ggtcgacctc
     1981 aacggaacaa gttaccacta caacgacctg gtgtctccac agtacctgaa cctcatcgca
     2041 aacctgtcgg gctgtaccgc ccaccggcgc gtgaacaact gctcggacat gtgcttccac
     2101 cagaagtacc ggacgcacga cggcacctgt aacaacctgc agcaccccat gtggggcgcc
     2161 tcgctgaccg ccttcgagcg cctgctgaaa tccgtgtacg agaatggctt caacacccct
     2221 cggggcatca acccccaccg actgtacaac gggcacgccc ttcccatgcc gcgcctggtg
     2281 tccaccaccc tgatcgggac ggagaccgtc acacccgacg agcagttcac ccacatgctg
     2341 atgcagtggg gccagttcct ggaccacgac ctcgactcca cggtggtggc cctgagccag
     2401 gcacgcttct ccgacggaca gcactgcagc aacgtgtgca gcaacgaccc cccctgcttc
     2461 tctgtcatga tcccccccaa tgactcccgg gccaggagcg gggcccgctg catgttcttc
     2521 gtgcgctcca gccctgtgtg cggcagcggc atgacttcgc tgctcatgaa ctccgtgtac
     2581 ccgcgggagc agatcaacca gctcacctcc tacatcgacg catccaacgt gtacgggagc
     2641 acggagcatg aggcccgcag catccgcgac ctggccagcc accgcggcct gctgcggcag
     2701 gtaagtatca aggttacaag acaggtttaa ggagaccaat aagctattat tcgtatagga
     2761 tactttatac gaagttatta agttatggag accggtgtgg cacgtgcgga ccgagcggcc
     2821 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     2881 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     2941 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     3001 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3061 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     3121 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3181 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3241 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3301 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3361 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     3421 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3481 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3541 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3601 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3661 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3721 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3781 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3841 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3901 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3961 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4021 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4081 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4141 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4201 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4261 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4321 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4381 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4441 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4501 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4561 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4621 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4681 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4741 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4801 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4861 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4921 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4981 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5041 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5101 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5161 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5221 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5281 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5341 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5401 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5461 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5521 ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca gctgcgcgct
     5581 cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
     5641 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcac
     5701 gcgtggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     5761 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     5821 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     5881 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     5941 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     6001 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     6061 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     6121 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     6181 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     6241 gcgcctaccg tacggccacc atgtaccctt atgacgtacc tgactatgct ggatctgcca
     6301 agcgctccag gggccccggg cgccgctgcc tgttggcgct cgtgctgttc tgcgcctggg
     6361 ggacgctggc cgtggtggcc cagaagccgg gcgcagggtg tccgagc
//
