LOCUS       sequence-489080-        5663 bp    DNA     circular SYN 09-OCT-2025
DEFINITION  5' AAVLINK plasmid for PCDH11X expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5663)
  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  JOURNAL   Unpublished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 5663)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 9, 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            "Unpublished"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'-PCDH11X
FEATURES             Location/Qualifiers
     source          1..5663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2105..2245
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2105..2234
                     /label=AAV2 ITR
     rep_origin      2320..2775
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2407..2426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2617..2638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2792..2811)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2911..2933
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2971..2989)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3057..3161
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3162..3230,3231..4022)
                     /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(3380..3399)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4193..4781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4682..4701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4843..4972
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        5029..5240
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5253..5521
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      5549..5558
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 acgtcctgat aggcgacttg ttgaaagacc ttaacttgtc gctgattcca aacaagtcct
       61 tgacaactgc tatgcagttc aagctagtgt acaagaccgg agatgtgcca ctgattcgaa
      121 ttgaagagga tactggtgag atcttcacta ctggcgctcg cattgatcgt gagaaattat
      181 gtgctggtat cccaagggat gagcattgct tttatgaagt ggaggttgcc attttgccgg
      241 atgaaatatt tagactggtt aagatacgtt ttctgataga agatataaat gataatgcac
      301 cattgttccc agcaacagtt atcaacatat caattccaga gaactcggct ataaactcta
      361 aatatactct cccagcggct gttgatcctg acgtaggaat aaacggagtt caaaactacg
      421 aactaattaa gagtcaaaac atttttggcc tcgatgtcat tgaaacacca gaaggagaca
      481 agatgccaca actgattgtt caaaaggagt tagataggga agagaaggat acctacgtga
      541 tgaaagtaaa ggttgaagat ggtggctttc ctcaaagatc cagtactgct attttgcaag
      601 tgagtgttac tgatacaaat gacaaccacc cagtctttaa ggagacagag attgaagtca
      661 gtataccaga aaatgctcct gtaggcactt cagtgacaca gctccatgcc acagatgctg
      721 acataggtga aaatgccaag atccacttct ctttcagcaa tctagtctcc aacattgcca
      781 ggagattatt tcacctcaat gccaccactg gacttatcac aatcaaagaa ccactggata
      841 gggaagaaac accaaaccac aagttactgg ttttggcaag tgatggtgga ttgatgccag
      901 caagagcaat ggtgctggta aatgttacag atgtcaatga taatgtccca tccattgaca
      961 taagatacat cgtcaatcct gtcaatgaca cagttgttct ttcagaaaat attccactca
     1021 acaccaaaat tgctctcata actgtgacgg ataaggatgc ggaccataat ggcagggtga
     1081 catgcttcac agatcatgaa atccctttca gattaaggcc agtattcagt aatcagttcc
     1141 tcctggagac tgcagcatat cttgactatg agtccacaaa agaatatgcc attaaattac
     1201 tggctgcaga tgctggcaaa cctcctttga atcagtcagc aatgctcttc atcaaagtga
     1261 aagatgaaaa tgacaatgct ccagttttca cccagtcttt cgtaactgtt tctattcctg
     1321 agaataactc tcctggcatc cagttgacga aagtaagtgc aatggatgca gacagtgggc
     1381 ctaatgctaa gatcaattac ctgctaggcc ctgatgctcc acctgaattc agcctggatt
     1441 gtcgtacagg catgctgact gtagtgaaga aactagatag agaaaaagag gataaatatt
     1501 tattcacaat tctggcaaaa gataacgggg taccaccctt aaccagcaat gtcacagtct
     1561 ttgtaagcat tattgatcag aatgacaata gcccagtttt cactcacaat gaatacaact
     1621 tctatgtccc agaaaacctt ccaaggcatg gtacagtagg actaatcact gtaactgatc
     1681 ctgattatgg agacaattct gcagttacgc tctccatttt agatgagaat gatgacttca
     1741 ccattgattc acaaactggt gtcatccgac caaatatttc atttgataga gaaaaacaag
     1801 aatcttacac tttctatgta aaggctgagg atggtggtag agtatcacgt tcttcaagtg
     1861 ccaaagtaac cataaatgtg gttgatgtca atgacaacaa accagttttc attgtccctc
     1921 cttccaactg ttcttatgaa ttggttctac cgtccactaa tccaggcaca gtggtctttc
     1981 aggtaagtat caaggttaca agacaggttt aaggagacca ataagctatt attcgtatag
     2041 gatactttat acgaagttat taagttatgg agaccggtgt ggcacgtgcg gaccgagcgg
     2101 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     2161 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     2221 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     2281 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     2341 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     2401 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2461 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2521 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2581 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2641 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     2701 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2761 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     2821 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     2881 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     2941 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     3001 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     3061 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3121 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3181 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3241 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3301 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3361 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3421 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3481 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3541 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3601 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3661 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3721 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3781 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3841 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3901 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3961 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4021 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4081 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4141 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4201 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4261 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4321 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4381 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4441 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4501 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4561 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4621 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4681 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4741 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4801 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg cagctgcgcg
     4861 ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
     4921 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
     4981 acgcgtgtcg actctagagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca
     5041 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga
     5101 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     5161 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     5221 ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc
     5281 gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct
     5341 gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg
     5401 gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc
     5461 tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat
     5521 ccaagctgtg accggcgcct accgtacggc caccatggac ttgttgtccg ggacgtacat
     5581 tttcgcggtc ctgctagcat gcgtggtgtt ccactctggc gcccaggaga aaaactacac
     5641 catccgagaa gaaatgccag aaa
//
