LOCUS       sequence-476869-        6229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' AAVLINK plasmid for CDC42BPB expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6229)
  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  JOURNAL   Unpublished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 6229)
  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            "Unpublished"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: pAAVLINKv1-pCALM1-Cre-3' CDC42BPB
FEATURES             Location/Qualifiers
     source          1..6229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     360..379
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1845..1910
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1975..1995)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2388..2528
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2388..2517
                     /label=AAV2 ITR
     rep_origin      2603..3058
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2690..2709)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2900..2921
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3075..3094)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3194..3216
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3254..3272)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3340..3444
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3445..3513,3514..4305)
                     /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(3663..3682)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4476..5064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4965..4984
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5126..5255
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             join(5402..6229,1..204)
                     /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"
ORIGIN
        1 gatgactctg gtcagagata cctggcctgg tctggacaca gtgcccgtgt cggagccgcg
       61 cgagatatgg cccgcgctgg agtttcaata ccggagatca tgcaagctgg tggctggacc
      121 aatgtaaata ttgtaatgaa ctatatccgt aacctggata gtgaaacagg ggcaatggtg
      181 cgcctgctgg aagatggcga ttaggtaagt atcaaggtta caagacaggt ttaaggagac
      241 caataactag tataacttcg tataggatac tttatagcaa tttatgaaac tgggcttgtc
      301 gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt
      361 tgcctttctc tccacaggtc gtgcatgttc ccttggaagc ctacgacagc tcgctgcctc
      421 tcatcaaggc catcctgaca gctgccatcg tggatgcaga caggattgca gtcggcctag
      481 aagaagggct ctatgtcata gaggtcaccc gagatgtgat cgtccgtgcc gctgactgta
      541 agaaggtaca ccagatcgag cttgctccca gggagaagat cgtaatcctc ctctgtggcc
      601 ggaaccacca tgtgcacctc tatccgtggt cgtcccttga tggagcggaa ggcagctttg
      661 acatcaagct tccggaaacc aaaggctgcc agctcatggc cacggccaca ctcaagagga
      721 actctggcac ctgcctgttt gtggccgtga aacggctgat cctttgctat gagatccaga
      781 gaacgaagcc attccacaga aagttcaatg agattgtggc tcccggcagc gtgcagtgcc
      841 tggcggtgct cagggacagg ctctgtgtgg gctacccttc tgggttctgc ctgctgagca
      901 tccaggggga cgggcagcct ctaaacctgg taaatcccaa tgacccctcg cttgcgttcc
      961 tctcacaaca gtcttttgat gccctttgtg ctgtggagct cgaaagcgag gagtacctgc
     1021 tttgcttcag ccacatggga ctgtacgtgg acccgcaagg ccggagggca cgcgcgcagg
     1081 agctcatgtg gcctgcggct cctgtcgcct gtagttgcag ccccacccac gtcacggtgt
     1141 acagcgagta tggcgtggac gtctttgatg tgcgcaccat ggagtgggtg cagaccatcg
     1201 gcctgcggag gataaggccc ctgaactctg aaggcaccct caacctcctc aactgcgagc
     1261 ctccacgctt gatctacttc aagagcaagt tctcgggagc ggttctcaac gtgccggaca
     1321 cctccgacaa cagcaagaag cagatgctgc gcaccaggag caaaaggcgg ttcgtcttca
     1381 aggtcccaga ggaagagaga ctgcagcaga ggcgagagat gcttagagac ccagaattga
     1441 gatccaaaat gatatccaac ccaaccaact tcaaccacgt ggcccacatg ggcccaggcg
     1501 acggcatgca ggtgctcatg gacctgcctc tgagtgctgt gcccccctcc caggaggaaa
     1561 ggccgggccc cgctcccacc aacctggctc gccagcctcc atccaggaac aagccctaca
     1621 tctcgtggcc ctcatcaggt ggatcggagc ctagcgtgac tgtgcctctg agaagtatgt
     1681 ctgatccaga ccaggacttt gacaaagagc ctgattcgga ctccaccaaa cactcaactc
     1741 catcgaatag ctccaacccc agcggcccac cgagccccaa ctccccccac aggagccagc
     1801 tccccctcga aggcctggag cagccggcct gtgacacctg tacagactac aaagaccatg
     1861 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag taactcgaga
     1921 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1981 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2041 tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca cggcggaact
     2101 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2161 cgtggtgttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca tctagcttta
     2221 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2281 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     2341 tttaaagatc tggagaccgg tgtggcacgt gcggaccgag cggccgcagg aacccctagt
     2401 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     2461 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     2521 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2581 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2641 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     2701 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2761 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     2821 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2881 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     2941 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     3001 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     3061 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     3121 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3181 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3241 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     3301 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3361 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3421 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3481 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3541 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3601 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3661 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3721 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3781 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3841 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3901 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3961 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4021 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4081 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4141 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4201 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4261 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4321 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4381 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4441 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4501 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4561 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4621 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4681 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4741 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4801 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4861 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4921 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4981 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5041 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5101 ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag
     5161 gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
     5221 agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgtc gctctgcgcg
     5281 aggcggctcc gccgcggcgg agggatacgg cgcaccatat atatatcgcg gggcgcagac
     5341 tcgcgctccg gcagtggtgc tgggagtgtc gtggacgccg tgccgttact cgtacgccac
     5401 catgtccaat ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg atgcaacgag
     5461 tgatgaggtt cgcaagaacc tgatggacat gttcagggat cgccaggcgt tttctgagca
     5521 tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca agttgaataa
     5581 ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat tatcttctat atcttcaggc
     5641 gcgcggtctg gcagtaaaaa ccatccagca acatttgggc cagctaaaca tgcttcatcg
     5701 tcggtccggg ctgccacgac caagtgacag caatgctgtt tcactggtta tgcggcggat
     5761 ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt tcgaacgcac
     5821 tgatttcgac caggttcgtt cactcatgga aaatagcgat cgctgccagg atatacgtaa
     5881 tctggcattt ctggggattg cttataacac cctgttacgt atagccgaaa ttgccaggat
     5941 cagggttaaa gatatctcac gtactgacgg tgggagaatg ttaatccata ttggcagaac
     6001 gaaaacgctg gttagcaccg caggtgtaga gaaggcactt agcctggggg taactaaact
     6061 ggtcgagcga tggatttccg tctctggtgt agctgatgat ccgaataact acctgttttg
     6121 ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc agccagctat caactcgcgc
     6181 cctggaaggg atttttgaag caactcatcg attgatttac ggcgctaag
//
