LOCUS       sequence-481049-        6743 bp    DNA     circular SYN 25-AUG-2025
DEFINITION  3' AAVLINK plasmid for SETD5 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6743)
  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  JOURNAL   Unpublished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 6743)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 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            "Unpublished"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..6743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     CDS             288..1319
                     /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             3451..3516
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3593..3613)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4006..4146
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4006..4135
                     /label=AAV2 ITR
     rep_origin      4221..4676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4308..4327)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4518..4539
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4693..4712)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4812..4834
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4872..4890)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4958..5062
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5063..5131,5132..5923)
                     /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(5281..5300)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6094..6682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6583..6602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtcgctc tgcgcgaggc ggctccgccg
      181 cggcggaggg atacggcgca ccatatatat atcgcggggc gcagactcgc gctccggcag
      241 tggtgctggg agtgtcgtgg acgccgtgcc gttactcgta cgccaccatg tccaatttac
      301 tgaccgtaca ccaaaatttg cctgcattac cggtcgatgc aacgagtgat gaggttcgca
      361 agaacctgat ggacatgttc agggatcgcc aggcgttttc tgagcatacc tggaaaatgc
      421 ttctgtccgt ttgccggtcg tgggcggcat ggtgcaagtt gaataaccgg aaatggtttc
      481 ccgcagaacc tgaagatgtt cgcgattatc ttctatatct tcaggcgcgc ggtctggcag
      541 taaaaactat ccagcaacat ttgggccagc taaacatgct tcatcgtcgg tccgggctgc
      601 cacgaccaag tgacagcaat gctgtttcac tggttatgcg gcggatccga aaagaaaacg
      661 ttgatgccgg tgaacgtgca aaacaggctc tagcgttcga acgcactgat ttcgaccagg
      721 ttcgttcact catggaaaat agcgatcgct gccaggatat acgtaatctg gcatttctgg
      781 ggattgctta taacaccctg ttacgtatag ccgaaattgc caggatcagg gttaaagata
      841 tctcacgtac tgacggtggg agaatgttaa tccatattgg cagaacgaaa acgctggtta
      901 gcaccgcagg tgtagagaag gcacttagcc tgggggtaac taaactggtc gagcgatgga
      961 tttccgtctc tggtgtagct gatgatccga ataactacct gttttgccgg gtcagaaaaa
     1021 atggtgttgc cgcgccatct gccaccagcc agctatcaac tcgcgccctg gaagggattt
     1081 ttgaagcaac tcatcgattg atttacggcg ctaaggatga ctctggtcag agatacctgg
     1141 cctggtctgg acacagtgcc cgtgtcggag ccgcgcgaga tatggcccgc gctggagttt
     1201 caataccgga gatcatgcaa gctggtggct ggaccaatgt aaatattgta atgaactata
     1261 tccgtaacct ggatagtgaa acaggggcaa tggtgcgcct gctggaagat ggcgattagg
     1321 taagtatcaa ggttacaaga caggtttaag gagaccaata actagtataa cttcgtatag
     1381 gatactttat agcaatttat gaaactgggc ttgtcgagac agagaagact cttgcgtttc
     1441 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac agggatgact
     1501 caaacatcat ctgtacccca agagactaga actcagcacc tataccaaag caatgagaat
     1561 agtagctctt ctagtatctg caaagacaat gcagacttgt tgagcccatt aaagaaatgg
     1621 aagtctcgct atctgatgga gcagaatgtc accaagttac ttcggcctct gtctccagtc
     1681 acaccacccc ctcccaattc aggctcaaag agtccccagc tggccacacc tggctcatct
     1741 cacccaggag aagaggagtg tcgaaatgga tacagcctca tgttttcacc agtcacatct
     1801 cttactactg ctagtcgctg caacactcct ctacagtttg agctttgtca ccgaaaagac
     1861 ctggatttgg caaaagtagg ataccttgac tccaacacta acagctgtgc tgatagacct
     1921 tccctactca actcaggtca ttctgacctg gctcctcatc cctccctcgg acccacttct
     1981 gagactggtt tcccaagcag aagtggagat ggacatcaga ccctcgtgag aaactcagac
     2041 caggcatttc ggacagagtt caacttgatg tatgcctact cccctttgaa tgctatgcct
     2101 cgagcagatg gactgtatcg aggatctcct ctagtggggg ataggaagcc tttacatttg
     2161 gatgggggat attgttcccc tgcagaagga ttttccagca gatatgaaca tggcttaatg
     2221 aaagacctct ctcgtggatc cttgtcacct ggtggtgaaa gggcctgtga aggagtccca
     2281 tctgcccccc agaacccacc acagaggaaa aaagtatccc tgctggagta ccgaaaacgg
     2341 aaacaagaag ctaaggaaaa ttctgctggt gggggaggtg actctgcaca gagcaaaagc
     2401 aagtctgcag gagctgggca aggcagcagt aactccgttt ccgacactgg tgcccatggt
     2461 gtgcagggat cctcagcccg aactccatct tcccctcaca aaaaattctc cccatctcat
     2521 tcctctatgt cccatttgga ggcggtaagc ccatcagatt ccagaggcac ttcttcatct
     2581 cactgcagac ctcaagagaa tatcagcagt aggtggatgg ttcccacatc agtagaacga
     2641 ctccgagaag gagggagcat ccccaaggtc ctccgaagca gcgtgagggt ggcccaaaag
     2701 ggagagccct ctcccacatg ggagagtaac atcacagaga aagactcaga ccctgcagat
     2761 ggagaaggcc cagagacatt aagctcagca ctctctaaag gagcaacagt ttacagccct
     2821 tccagataca gctaccagct cctgcagtgt gatagtcctc ggacagaatc acaaagcctc
     2881 cttcagcaga gttcctcccc cttcagagga catcctacac agtctccagg atacagttat
     2941 cgaactactg cactgagacc tggaaacccc ccctctcacg gttcttcaga atcatccctc
     3001 tcttccacgt cctattccag ccccgcccac cctgtgtcca cagactcgtt ggccccattt
     3061 acggggacac cagggtattt tagcagccag ccacattctg gaaacagcac tggcagcaat
     3121 cttccaagga ggagctgccc ttctagtgct gctagcccta ccctgcaggg accctcagac
     3181 tcgccaacct cagattcagt ttctcagtcc agcacaggaa ctctgagttc cacctccttt
     3241 cctcagaact ctaggtcgtc attgccatca gacttacgga ctatcagtct gcccagtgct
     3301 gggcagtcag ctgtctacca ggcctccagg gtatctgcgg tttccaattc acagcactac
     3361 ccacaccgtg ggagtggggg tgtgcaccag taccgactcc agccactgca agggtcagga
     3421 gtcaagactc agacgggact ttcctgtaca gactacaaag accatgacgg tgattataaa
     3481 gatcatgaca tcgactacaa ggatgacgat gacaagtaag aattctgatg actcgagata
     3541 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3601 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3661 tggctttcat tttctcctcc ttgtataaat cctggttagt tcttgccacg gcggaactca
     3721 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3781 tggtgtttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatc tagctttatt
     3841 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3901 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     3961 taaagatctg gagaccggtg tggcacgtgc ggaccgagcg gccgcaggaa cccctagtga
     4021 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4081 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4141 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4201 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4261 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4321 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4381 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4441 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4501 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4561 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4621 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4681 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4741 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4801 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4861 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4921 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4981 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5041 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5101 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5161 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5221 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5281 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5341 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5401 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5461 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5521 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5581 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5641 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5701 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5761 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5821 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5881 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5941 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6001 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6061 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6121 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6181 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6241 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6301 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6361 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6421 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6481 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6541 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6601 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6661 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6721 ttgctggcct tttgctcaca tgt
//
