LOCUS       sequence-476855-        6610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' AAVLINK plasmid for IQGAP3 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6610)
  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  JOURNAL   Unpublished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 6610)
  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            "Unpublished"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' IQGAP3
FEATURES             Location/Qualifiers
     source          1..6610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     169..188
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2023..2088
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2165..2185)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2578..2718
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2578..2707
                     /label=AAV2 ITR
     rep_origin      2793..3248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2880..2899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3090..3111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3265..3284)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3384..3406
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3444..3462)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3530..3634
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3635..3703,3704..4495)
                     /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(3853..3872)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4666..5254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5155..5174
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5316..5445
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             join(5592..6610,1..13)
                     /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 agatggcgat taggtaagta tcaaggttac aagacaggtt taaggagacc aataactagt
       61 ataacttcgt ataggatact ttatagcaat ttatgaaact gggcttgtcg agacagagaa
      121 gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt gcctttctct
      181 ccacaggtgg agcagcccca ggacgtggtg acaggcaacc caacagtggt gaggctggtg
      241 gtgagattct accgtaatgg gcggggacag agtgccctgc aggagattct gggcaaggtt
      301 atccaggatg tgctagaaga caaagtgctc agcgtccaca cagaccctgt ccacctctat
      361 aagaactgga tcaaccagac tgaggcccag acagggcagc gcagccatct cccatatgat
      421 gtcaccccgg agcaggcctt gagccacccc gaggtccaga gacgactgga catcgcccta
      481 cgcaacctcc tcgccatgac tgataagttc cttttagcca tcacctcatc tgtggaccaa
      541 attccgtatg ggatgcgata tgtggccaaa gtcctgaagg caactctggc agagaaattc
      601 cctgacgcca cagacagcga ggtctataag gtggtcggga acctcctgta ctaccgcttc
      661 ctgaacccag ctgtggtggc tcctgacgcc ttcgacattg tggccatggc agctggtgga
      721 gccctggctg ccccccagcg ccatgccctg ggggctgtgg ctcagctcct acagcacgct
      781 gcggctggca aggccttctc tgggcagagc cagcacctac gggtcctgaa tgactatctg
      841 gaggaaacac acctcaagtt caggaagttc atccatagag cctgccaggt gccagagcca
      901 gaggagcgtt ttgcagtgga cgagtactca gacatggtgg ctgtggccaa acccatggtg
      961 tacatcaccg tgggggagct ggtcaacacg cacaggctgt tgctggagca ccaggactgc
     1021 attgcccctg atcaccaaga ccccctgcat gagctcctgg aggatcttgg ggagctgccc
     1081 accatccctg accttattgg tgagagcatc gctgcagatg ggcacacgga cctgagcaag
     1141 ctagaagtgt ccctgacgct gaccaacaag tttgaaggac tagaggcaga tgctgatgac
     1201 tccaacaccc gtagcctgct tctgagcacc aagcagctgt tggccgatat catacagttc
     1261 catcctgggg acaccctcaa ggagatcctg tccctctcgg cttccagaga gcaagaagca
     1321 gcccacaagc agctgatgag ccgacgccag gcctgtacag cccagacacc ggagccactg
     1381 cgacgacacc gctcactgac agctcactcc ctcctgccac tggcagagaa gcagcggcgc
     1441 gtcctgcgga acctacgccg acttgaagcc ctggggttgg tcagcgccag aaatggctac
     1501 caggggctag tggacgagct ggccaaggac atccgcaacc agcacagaca caggcacagg
     1561 cggaaggcag agctggtgaa gctgcaggcc acattacagg gcctgagcac taagaccacc
     1621 ttctatgagg agcagggtga ctactacagc cagtacatcc gggcctgcct ggaccacctg
     1681 gcccccgact ccaagagttc tgggaagggg aagaagcagc cttctcttca ttacactgct
     1741 gctcagctcc tggaaaaggg tgtcttggtg gaaattgaag atcttcccgc ctctcacttc
     1801 agaaacgtca tctttgacat cacgccggga gatgaggcag gaaagtttga agtaaatgcc
     1861 aagttcctgg gtgtggacat ggagcgattt cagcttcact atcaggatct cctgcagctc
     1921 cagtatgagg gtgtggctgt catgaaactc ttcaacaagg ccaaagtcaa tgtcaacctt
     1981 ctcatcttcc tcctcaacaa gaagtttttg cggaagtgta cagactacaa agaccatgac
     2041 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta agaattctga
     2101 tgactcgaga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2161 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2221 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggtta gttcttgcca
     2281 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2341 ctgacaattc cgtggtgttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2401 tctagcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2461 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     2521 tgggaggttt tttaaagatc tggagaccgg tgtggcacgt gcggaccgag cggccgcagg
     2581 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     2641 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     2701 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2761 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     2821 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     2881 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2941 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3001 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3061 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3121 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     3181 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3241 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3301 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3361 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3421 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3481 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3541 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3601 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3661 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3721 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3781 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3841 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3901 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3961 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4021 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4081 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4141 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4201 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4261 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4321 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4381 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4441 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4501 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4561 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4621 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4681 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4741 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4801 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4861 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4921 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4981 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5041 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5101 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5161 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5221 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5281 gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc
     5341 gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
     5401 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgtc
     5461 gctctgcgcg aggcggctcc gccgcggcgg agggatacgg cgcaccatat atatatcgcg
     5521 gggcgcagac tcgcgctccg gcagtggtgc tgggagtgtc gtggacgccg tgccgttact
     5581 cgtacgccac catgtccaat ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg
     5641 atgcaacgag tgatgaggtt cgcaagaacc tgatggacat gttcagggat cgccaggcgt
     5701 tttctgagca tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca
     5761 agttgaataa ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat tatcttctat
     5821 atcttcaggc gcgcggtctg gcagtaaaaa ccatccagca acatttgggc cagctaaaca
     5881 tgcttcatcg tcggtccggg ctgccacgac caagtgacag caatgctgtt tcactggtta
     5941 tgcggcggat ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt
     6001 tcgaacgcac tgatttcgac caggttcgtt cactcatgga aaatagcgat cgctgccagg
     6061 atatacgtaa tctggcattt ctggggattg cttataacac cctgttacgt atagccgaaa
     6121 ttgccaggat cagggttaaa gatatctcac gtactgacgg tgggagaatg ttaatccata
     6181 ttggcagaac gaaaacgctg gttagcaccg caggtgtaga gaaggcactt agcctggggg
     6241 taactaaact ggtcgagcga tggatttccg tctctggtgt agctgatgat ccgaataact
     6301 acctgttttg ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc agccagctat
     6361 caactcgcgc cctggaaggg atttttgaag caactcatcg attgatttac ggcgctaagg
     6421 atgactctgg tcagagatac ctggcctggt ctggacacag tgcccgtgtc ggagccgcgc
     6481 gagatatggc ccgcgctgga gtttcaatac cggagatcat gcaagctggt ggctggacca
     6541 atgtaaatat tgtaatgaac tatatccgta acctggatag tgaaacaggg gcaatggtgc
     6601 gcctgctgga
//
