LOCUS       sequence-491767-        6732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3' AAVLINK plasmid for SETD5 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6732)
  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  JOURNAL   Unpublished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 6732)
  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            "Unpublished"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'-SETD5-FLAG
FEATURES             Location/Qualifiers
     source          1..6732
                     /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"
     CDS             3440..3505
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3582..3602)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3995..4135
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3995..4124
                     /label=AAV2 ITR
     rep_origin      4210..4665
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4297..4316)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4507..4528
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4682..4701)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4801..4823
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4861..4879)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4947..5051
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5052..5120,5121..5912)
                     /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(5270..5289)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6083..6671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6572..6591
                     /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 gggatgactc aaacatcatc
     1501 tgtaccccaa gagactagaa ctcagcacct ataccaaagc aatgagaata gtagctcttc
     1561 tagtatctgc aaagacaatg cagacttgtt gagcccatta aagaaatgga agtctcgcta
     1621 tctgatggag cagaatgtca ccaagttact tcggcctctg tctccagtca caccaccccc
     1681 tcccaattca ggctcaaaga gtccccagct ggccacacct ggctcatctc acccaggaga
     1741 agaggagtgt cgaaatggat acagcctcat gttttcacca gtcacatctc ttactactgc
     1801 tagtcgctgc aacactcctc tacagtttga gctttgtcac cgaaaagacc tggatttggc
     1861 aaaagtagga taccttgact ccaacactaa cagctgtgct gatagacctt ccctactcaa
     1921 ctcaggtcat tctgacctgg ctcctcatcc ctccctcgga cccacttctg agactggttt
     1981 cccaagcaga agtggagatg gacatcagac cctcgtgaga aactcagacc aggcatttcg
     2041 gacagagttc aacttgatgt atgcctactc ccctttgaat gctatgcctc gagcagatgg
     2101 actgtatcga ggatctcctc tagtggggga taggaagcct ttacatttgg atgggggata
     2161 ttgttcccct gcagaaggat tttccagcag atatgaacat ggcttaatga aagacctctc
     2221 tcgtggatcc ttgtcacctg gtggtgaaag ggcctgtgaa ggagtcccat ctgcccccca
     2281 gaacccacca cagaggaaaa aagtatccct gctggagtac cgaaaacgga aacaagaagc
     2341 taaggaaaat tctgctggtg ggggaggtga ctctgcacag agcaaaagca agtctgcagg
     2401 agctgggcaa ggcagcagta actccgtttc cgacactggt gcccatggtg tgcagggatc
     2461 ctcagcccga actccatctt cccctcacaa aaaattctcc ccatctcatt cctctatgtc
     2521 ccatttggag gcggtaagcc catcagattc cagaggcact tcttcatctc actgcagacc
     2581 tcaagagaat atcagcagta ggtggatggt tcccacatca gtagaacgac tccgagaagg
     2641 agggagcatc cccaaggtcc tccgaagcag cgtgagggtg gcccaaaagg gagagccctc
     2701 tcccacatgg gagagtaaca tcacagagaa agactcagac cctgcagatg gagaaggccc
     2761 agagacatta agctcagcac tctctaaagg agcaacagtt tacagccctt ccagatacag
     2821 ctaccagctc ctgcagtgtg atagtcctcg gacagaatca caaagcctcc ttcagcagag
     2881 ttcctccccc ttcagaggac atcctacaca gtctccagga tacagttatc gaactactgc
     2941 actgagacct ggaaaccccc cctctcacgg ttcttcagaa tcatccctct cttccacgtc
     3001 ctattccagc cccgcccacc ctgtgtccac agactcgttg gccccattta cggggacacc
     3061 agggtatttt agcagccagc cacattctgg aaacagcact ggcagcaatc ttccaaggag
     3121 gagctgccct tctagtgctg ctagccctac cctgcaggga ccctcagact cgccaacctc
     3181 agattcagtt tctcagtcca gcacaggaac tctgagttcc acctcctttc ctcagaactc
     3241 taggtcgtca ttgccatcag acttacggac tatcagtctg cccagtgctg ggcagtcagc
     3301 tgtctaccag gcctccaggg tatctgcggt ttccaattca cagcactacc cacaccgtgg
     3361 gagtgggggt gtgcaccagt accgactcca gccactgcaa gggtcaggag tcaagactca
     3421 gacgggactt tcctgtacag actacaaaga ccatgacggt gattataaag atcatgacat
     3481 cgactacaag gatgacgatg acaagtaaga attctgatga ctcgagataa tcaacctctg
     3541 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3601 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3661 ttctcctcct tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc
     3721 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgtttatt
     3781 tgtgaaattt gtgatgctat tgctttattt gtaaccatct agctttattt gtgaaatttg
     3841 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3901 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagatctgg
     3961 agaccggtgt ggcacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     4021 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     4081 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     4141 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     4201 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4261 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     4321 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4381 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4441 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4501 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     4561 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     4621 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     4681 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4741 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4801 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4861 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4921 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4981 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5041 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5101 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5161 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5221 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5281 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5341 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5401 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5461 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5521 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5581 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5641 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5701 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5761 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5821 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5881 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5941 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6001 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6061 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6121 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6181 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6241 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6301 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6361 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6421 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6481 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6541 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6601 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6661 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6721 ttgctcacat gt
//
