LOCUS       sequence-491861-        6748 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  3' AAVLINK plasmid for PCDH11X expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6748)
  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  JOURNAL   Unpublished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 6748)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAVLINKv1-pCALM1-Cre-3'-PCDH11X-HA
FEATURES             Location/Qualifiers
     source          1..6748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             277..1308
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     primer_bind     1464..1483
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3447..3473
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3477..3503
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3507..3533
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3598..3618)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4011..4151
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4011..4140
                     /label=AAV2 ITR
     rep_origin      4226..4681
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4313..4332)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4523..4544
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4698..4717)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4817..4839
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4877..4895)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4963..5067
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5068..5136,5137..5928)
                     /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(5286..5305)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6099..6687
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6588..6607
                     /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 gcgtcgctct gcgcgaggcg gctccgccgc ggcggaggga
      181 tacggcgcac catatatata tcgcggggcg cagactcgcg ctccggcagt ggtgctggga
      241 gtgtcgtgga cgccgtgccg ttactcgtac gccaccatgt ccaatttact gaccgtacac
      301 caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg
      361 gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct tctgtccgtt
      421 tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc cgcagaacct
      481 gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt aaaaaccatc
      541 cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc acgaccaagt
      601 gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt
      661 gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc
      721 atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg gattgcttat
      781 aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat ctcacgtact
      841 gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag caccgcaggt
      901 gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat ttccgtctct
      961 ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa tggtgttgcc
     1021 gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt tgaagcaact
     1081 catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc ctggtctgga
     1141 cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc aataccggag
     1201 atcatgcaag ctggtggctg gaccaatgta aatattgtaa tgaactatat ccgtaacctg
     1261 gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattaggt aagtatcaag
     1321 gttacaagac aggtttaagg agaccaataa ctagtataac ttcgtatagg atactttata
     1381 gcaatttatg aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     1441 tattggtctt actgacatcc actttgcctt tctctccaca ggtaattgct gttgacaatg
     1501 acactggcat gaatgcagag gttcgttaca gcattgtagg aggaaacaca agagatctgt
     1561 ttgcaatcga ccaagaaaca ggcaacataa cattgatgga gaaatgtgat gttacagacc
     1621 ttggtttaca cagagtgttg gtcaaagcta atgacttagg acagcctgat tctctcttca
     1681 gtgttgtaat tgtcaatctg ttcgtgaatg agtcggtgac caatgctaca ctgattaatg
     1741 aactggtgcg caaaagcact gaagcaccag tgaccccaaa tactgagata gctgatgtat
     1801 cctcaccaac tagtgactat gtcaagatcc tggttgcagc tgttgctggc accataactg
     1861 tcgttgtagt tattttcatc actgctgtag taagatgtcg ccaggcacca caccttaagg
     1921 ctgctcagaa aaacaagcag aattctgaat gggctacccc aaacccagaa aacaggcaga
     1981 tgataatgat gaagaaaaag aaaaagaaga agaagcattc ccctaagaac ttgctgctta
     2041 attttgtcac tattgaagaa actaaggcag atgatgttga cagtgatgga aacagagtca
     2101 cactagacct tcctattgat ctagaagagc aaacaatggg aaagtacaat tgggtaacta
     2161 cacctactac tttcaagccc gacagccctg atttggcccg acactacaaa tctgcctctc
     2221 cacagcctgc cttccaaatt cagcctgaaa ctcccctgaa ttcgaagcac cacatcatcc
     2281 aagaactgcc tctcgataac acctttgtgg cctgtgactc tatctccaag tgttcctcaa
     2341 gcagttcaga tccctacagc gtttctgact gtggctatcc agtgacgacc ttcgaggtac
     2401 ctgtgtccgt acacaccaga ccgccaatga aggaggttgt gcgatcttgc acccccatga
     2461 aagagtctac aactatggag atctggattc atccccaacc acagcggaaa tctgaaggga
     2521 aagtggcagg aaagtcccag cggcgtgtca catttcacct gccagaaggc tctcaggaaa
     2581 gcagcagtga tggtggactg ggagaccatg atgcaggcag ccttaccagc acatctcatg
     2641 gcctgcccct tggctatcct caggaggagt actttgatcg tgctacaccc agcaatcgca
     2701 ctgaagggga tggcaactcc gatcctgaat ctactttcat acctggacta aagaaagctg
     2761 cagaaataac tgttcaacca actgtggaag aggcctctga caactgcact caagaatgtc
     2821 tcatctatgg ccattctgat gcctgctgga tgccggcatc tctggatcat tccagctctt
     2881 cgcaagcaca ggcctctgct ctatgccaca gcccaccact gtcacaggcc tctactcagc
     2941 accacagccc acgagtgaca cagaccattg ctctctgcca cagccctcca gtgacacaga
     3001 ccatcgcatt gtgccacagc ccaccaccga tacaggtgtc tgctctccac cacagtcctc
     3061 ctctagtgca ggctactgca cttcaccaca gcccaccatc agcacaggcc tcagccctct
     3121 gctacagccc tcctttagca caggctgctg caatcagcca cagctctcct ctgccacagg
     3181 ttattgccct ccatcgtagt caggcccaat catcagtcag tttgcagcaa ggttgggtgc
     3241 aaggtgctga tgggctatgc tctgttgatc agggagtgca aggtagtgca acatctcagt
     3301 tttacaccat gtctgaaaga cttcatccca gtgatgattc aattaaagtc attcctttga
     3361 caaccttcac tccacgccaa caggccagac cgtccagagg tgattccccc attatggaag
     3421 aacatccctt gggaggtgga tctggttacc cgtatgatgt tccggattac gctggctatc
     3481 cctacgacgt gcccgactat gccgggtacc cctatgacgt cccagactac gcataactcg
     3541 agataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3601 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3661 ccgtatggct ttcattttct cctccttgta taaatcctgg ttagttcttg ccacggcgga
     3721 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3781 ttccgtggtg tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccatctagct
     3841 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     3901 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     3961 ttttttaaag atctggagac cggtgtggca cgtgcggacc gagcggccgc aggaacccct
     4021 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4081 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4141 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4201 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4261 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
     4321 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4381 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4441 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4501 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
     4561 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
     4621 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     4681 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4741 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4801 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4861 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4921 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4981 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5041 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5101 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5161 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5221 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5281 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5341 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5401 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5461 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5521 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5581 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5641 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5701 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5761 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5821 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5881 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5941 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6001 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6061 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6121 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6181 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6241 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6301 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6361 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6421 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6481 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6541 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6601 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6661 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6721 gccttttgct ggccttttgc tcacatgt
//
