LOCUS       sequence-476878-        6361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' AAVLINK plasmid for AHDC1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6361)
  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  JOURNAL   Unpublished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 6361)
  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            "Unpublished"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' AHDC1
FEATURES             Location/Qualifiers
     source          1..6361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1960..2025
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2090..2110)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2503..2643
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2503..2632
                     /label=AAV2 ITR
     rep_origin      2718..3173
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2805..2824)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3015..3036
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3190..3209)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3309..3331
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3369..3387)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3455..3559
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3560..3628,3629..4420)
                     /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(3778..3797)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4591..5179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5080..5099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5241..5370
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             join(5517..6361,1..187)
                     /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 atacctggcc tggtctggac acagtgcccg tgtcggagcc gcgcgagata tggcccgcgc
       61 tggagtttca ataccggaga tcatgcaagc tggtggctgg accaatgtaa atattgtaat
      121 gaactatatc cgtaacctgg atagtgaaac aggggcaatg gtgcgcctgc tggaagatgg
      181 cgattaggta agtatcaagg ttacaagaca ggtttaagga gaccaataac tagtataact
      241 tcgtatagga tactttatag caatttatga aactgggctt gtcgagacag agaagactct
      301 tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag
      361 ggcaccacag cctctgccac ctctgcagcc tctgccgcct cctcctcctc ctcctccttc
      421 cagccctcgc ccgagaactg tcggcagttt gcgggggctt ctcagtggcc tttccggcag
      481 ggctatggag gcctggactg ggcctcagag gcctttagtc agctctacaa tcccagtttt
      541 gactgccacg tcagcgagcc caacgtgatc ctggacatct ccaactacac accgcagaag
      601 gtgaagcagc agacggctgt gtcggagacc ttctctgagt catcctccga cagcacccag
      661 ttcaatcagc cggttggtgg cggggggttt cggcgtgcca acagcgaggc ctcaagtagt
      721 gagggccagt cgagcctgtc cagcctggag aaactgatga tggactggaa cgaggcatca
      781 tctgcccccg gctacaactg gaaccagagt gtcctctttc agagtagctc caagccgggc
      841 cgtggacggc ggaagaaggt ggacctgttc gaggcctcac atctgggctt cccgacatcc
      901 gcctctgccg ctgcctcagg ctacccatcc aaacggagca ctgggccccg gcagccgcga
      961 ggtggacggg gcggtggggc ctgctcagcc aagaaggagc ggggtggcgc agcggccaaa
     1021 gccaagttca tccccaagcc acagccagtc aacccactgt tccaggacag tcctgacctc
     1081 ggcctggact actatagcgg ggacagcagc atgtcaccac tgccctcaca gtcgagggcc
     1141 ttcggcgtgg gagagcgaga cccctgtgac ttcataggac cctactccat gaacccgtcc
     1201 acgccttccg atggcacctt tggccaaggc ttccactgcg actcgcccag cctgggtgct
     1261 cccgagcttg atggcaagca tttcccacct ctggcccacc cacccacggt gtttgacgcc
     1321 ggcctgcaga aggcatactc gcccacctgc tcgcctacac tgggcttcaa ggaagagctg
     1381 cggccaccgc ccacaaagct ggctgcctgc gagcccctca agcatggact ccagggggcc
     1441 agcctgggcc acgcagctgc agcccaggcc cacctgagct gccgggacct gccgctgggc
     1501 cagccccact acgattcccc cagctgcaag ggcacagcct attggtaccc tccaggctca
     1561 gctgcccgca gcccgcccta tgaaggcaag gtgggtacag ggctgctggc tgacttcctg
     1621 ggcaggacgg aggccgcgtg cctcagtgcc cctcacctgg ctagcccacc agccacgccc
     1681 aaggccgaca aggagccact ggaaatggcc cggccccctg gcccaccccg tggccctgct
     1741 gcagccgctg ctggctatgg ctgcccactc cttagtgact tgaccctgtc ccccgtgccg
     1801 agggactcgc tgctgcccct gcaggacacc gcctacaggt acccaggctt tatgccccag
     1861 gcgcatcctg gcctgggtgg gggccccaag agcggcttcc tggggcccat ggcggaacct
     1921 caccccgagg acacattcac cgtcacatcc ctgtgtacag actacaaaga ccatgacggt
     1981 gattataaag atcatgacat cgactacaag gatgacgatg acaagtaact cgagataatc
     2041 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2101 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2161 ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg gaactcatcg
     2221 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2281 tgtttatttg tgaaatttgt gatgctattg ctttatttgt aaccatctag ctttatttgt
     2341 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2401 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     2461 agatctggag accggtgtgg cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     2521 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     2581 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     2641 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     2701 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2761 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     2821 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2881 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     2941 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3001 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     3061 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     3121 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     3181 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     3241 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3301 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3361 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     3421 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3481 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3541 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3601 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3661 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3721 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3781 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3841 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3901 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3961 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4021 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4081 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4141 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4201 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4261 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4321 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4381 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4441 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4501 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4561 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4621 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4681 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4741 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4801 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4861 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4921 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4981 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5041 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5101 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5161 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5221 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     5281 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     5341 gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtcgctct gcgcgaggcg
     5401 gctccgccgc ggcggaggga tacggcgcac catatatata tcgcggggcg cagactcgcg
     5461 ctccggcagt ggtgctggga gtgtcgtgga cgccgtgccg ttactcgtac gccaccatgt
     5521 ccaatttact gaccgtacac caaaatttgc ctgcattacc ggtcgatgca acgagtgatg
     5581 aggttcgcaa gaacctgatg gacatgttca gggatcgcca ggcgttttct gagcatacct
     5641 ggaaaatgct tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg aataaccgga
     5701 aatggtttcc cgcagaacct gaagatgttc gcgattatct tctatatctt caggcgcgcg
     5761 gtctggcagt aaaaaccatc cagcaacatt tgggccagct aaacatgctt catcgtcggt
     5821 ccgggctgcc acgaccaagt gacagcaatg ctgtttcact ggttatgcgg cggatccgaa
     5881 aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt
     5941 tcgaccaggt tcgttcactc atggaaaata gcgatcgctg ccaggatata cgtaatctgg
     6001 catttctggg gattgcttat aacaccctgt tacgtatagc cgaaattgcc aggatcaggg
     6061 ttaaagatat ctcacgtact gacggtggga gaatgttaat ccatattggc agaacgaaaa
     6121 cgctggttag caccgcaggt gtagagaagg cacttagcct gggggtaact aaactggtcg
     6181 agcgatggat ttccgtctct ggtgtagctg atgatccgaa taactacctg ttttgccggg
     6241 tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca gctatcaact cgcgccctgg
     6301 aagggatttt tgaagcaact catcgattga tttacggcgc taaggatgac tctggtcaga
     6361 g
//
