pAAVLINKv1-pCALM1-Cre-3'-MSH6-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-MSH6-FLAG                6990 bp DNA     circular SYN 18-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6990)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported May 22, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    12..130
                     /label=AAV2 ITR
     promoter        145..265
                     /label=pCALM1
     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"
     intron          1309..1349
                     /label=Splicing Donor
     protein_bind    1356..1389
                     /label=lox2272-JTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          1390..1481
                     /label=Splicing Acceptor
                     /note="Splicing acceptor"
     CDS             1482..3703
                     /codon_start=1
                     /label=MSH6
                     /translation="V*YPAPSFGMHPKL*ELSLRKNILGKS*VMALG*CYPRCLKV*LQ
                     SLIPLG*HQERKVNWPSLL*VVVSSTSKNALLIRSFYQWLILKNIFPWILTQSALQDLV
                     LSSPKPINEWC*MQ*H*TTWRFF*MEQMVLLKEPY*RGLILAILLLVSGS*SNGFVPHS
                     VTIMLLMIV*MP*KTSWLCLTKSPKL*SF*RSFQILRGYSVKFIMLGLP*RVRTTQTAG
                     L*CMKKLHTARRRLLIFFLLWKDSK*CVKL*GSWKKLLMVLSLKSLSRSSLCRQKILKV
                     VFLI*L*N*TDGIQPLTMKRLERLDLLLPKQALTLIMTKLLLT*EKMNRASWNT*RNSA
                     TELAVGP*SIGGLVGTVTSWKFLRISPLAICQKNTS*NLPRRAVNDTGPKLLKRSWLIS
                     *MLKNGGMYH*RTACGDCSITLIKITRTGSLL*SVSQCWMFYCAWLTIVEGVMVLCVAQ
                     *FCCRKIPPPS*SLKDHAILALRRLFLEMILFLMTF**AVRKRSRKMAKPIVCLLLDQI
                     WGASLRL*DRLAY*L*WPRWVVTSLLKCAGSHQLIECLLDLVPQTE*CQVKVHFLLN*V
                     KLPAYSCMQQHILWCLWMN*EEVLQHLMGRQ*QMQLLKNLLRL*NVVHYFQLTTIH**K
                     IILKMLLCA*DIWHAW*KMNVKTPARRLLRSSINSLRELVLKAMALMQQGLLISQRKLF
                     KRDIEKQENLRR*ISHYDYFGKFAWLVKGQL*MLKLSINC*L*LRN"
     misc_feature    2473..2478
                     /label=SDS
     CDS             3710..3775
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3779..4210
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3923..3928
                     /label=SDS
     repeat_region   4253..4393
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4253..4382
                     /label=AAV2 ITR
     rep_origin      4468..4923
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5205..5309
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5310..6170
                     /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"
     rep_origin      6341..6929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 ggtctgatac ccggctccca
     1501 gttttgggat gcatccaaaa ctttgagaac tctccttgag gaagaatatt ttagggaaaa
     1561 gctaagtgat ggcattgggg tgatgttacc ccaggtgctt aaaggtatga cttcagagtc
     1621 tgattccatt gggttgacac caggagagaa aagtgaattg gccctctctg ctctaggtgg
     1681 ttgtgtcttc tacctcaaaa aatgccttat tgatcaggag cttttatcaa tggctaattt
     1741 tgaagaatat attcccttgg attctgacac agtcagcact acaagatctg gtgctatctt
     1801 caccaaagcc tatcaacgaa tggtgctaga tgcagtgaca ttaaacaact tggagatttt
     1861 tctgaatgga acaaatggtt ctactgaagg aaccctacta gagagggttg atacttgcca
     1921 tactcctttt ggtaagcggc tcctaaagca atggctttgt gccccactct gtaaccatta
     1981 tgctattaat gatcgtctag atgccataga agacctcatg gttgtgcctg acaaaatctc
     2041 cgaagttgta gagcttctaa agaagcttcc agatcttgag aggctactca gtaaaattca
     2101 taatgttggg tctcccctga agagtcagaa ccacccagac agcagggcta taatgtatga
     2161 agaaactaca tacagcaaga agaagattat tgattttctt tctgctctgg aaggattcaa
     2221 agtaatgtgt aaaattatag ggatcatgga agaagttgct gatggtttta agtctaaaat
     2281 ccttaagcag gtcatctctc tgcagacaaa aaatcctgaa ggtcgttttc ctgatttgac
     2341 tgtagaattg aaccgatggg atacagcctt tgaccatgaa aaggctcgaa agactggact
     2401 tattactccc aaagcaggct ttgactctga ttatgaccaa gctcttgctg acataagaga
     2461 aaatgaacag agcctcctgg aatacctaga gaaacagcgc aacagaattg gctgtaggac
     2521 catagtctat tgggggattg gtaggaaccg ttaccagctg gaaattcctg agaatttcac
     2581 cactcgcaat ttgccagaag aatacgagtt gaaatctacc aagaagggct gtaaacgata
     2641 ctggaccaaa actattgaaa agaagttggc taatctcata aatgctgaag aacggaggga
     2701 tgtatcattg aaggactgca tgcggcgact gttctataac tttgataaaa attacaagga
     2761 ctggcagtct gctgtagagt gtatcgcagt gttggatgtt ttactgtgcc tggctaacta
     2821 tagtcgaggg ggtgatggtc ctatgtgtcg cccagtaatt ctgttgccgg aagatacccc
     2881 ccccttctta gagcttaaag gatcacgcca tccttgcatt acgaagactt tttttggaga
     2941 tgattttatt cctaatgaca ttctaatagg ctgtgaggaa gaggagcagg aaaatggcaa
     3001 agcctattgt gtgcttgtta ctggaccaaa tatggggggc aagtctacgc ttatgagaca
     3061 ggctggctta ttagctgtaa tggcccagat gggttgttac gtccctgctg aagtgtgcag
     3121 gctcacacca attgatagag tgtttactag acttggtgcc tcagacagaa taatgtcagg
     3181 tgaaagtaca ttttttgttg aattaagtga aactgccagc atactcatgc atgcaacagc
     3241 acattctctg gtgcttgtgg atgaattagg aagaggtact gcaacatttg atgggacggc
     3301 aatagcaaat gcagttgtta aagaacttgc tgagactata aaatgtcgta cattattttc
     3361 aactcactac cattcattag tagaagatta ttctcaaaat gttgctgtgc gcctaggaca
     3421 tatggcatgc atggtagaaa atgaatgtga agaccccagc caggagacta ttacgttcct
     3481 ctataaattc attaagggag cttgtcctaa aagctatggc tttaatgcag caaggcttgc
     3541 taatctccca gaggaagtta ttcaaaaggg acatagaaaa gcaagagaat ttgagaagat
     3601 gaatcagtca ctacgattat ttcgggaagt ttgcctggct agtgaaaggt caactgtaga
     3661 tgctgaagct gtccataaat tgctgacttt gattaaggaa ttatgtacag actacaaaga
     3721 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtaact
     3781 cgagataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3841 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3901 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttagttct tgccacggcg
     3961 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4021 aattccgtgg tgtttatttg tgaaatttgt gatgctattg ctttatttgt aaccatctag
     4081 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4141 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     4201 ggttttttaa agatctggag accggtgtgg cacgtgcgga ccgagcggcc gcaggaaccc
     4261 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4321 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4381 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4441 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     4501 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     4561 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4621 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4681 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4741 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     4801 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     4861 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     4921 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     4981 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     5041 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     5101 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     5161 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5221 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5281 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5341 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5401 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5461 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5521 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5581 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5641 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5701 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5761 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5821 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5881 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5941 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6001 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6061 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6121 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6181 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6241 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6301 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6361 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6421 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6481 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6541 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6601 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6661 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6721 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6781 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6841 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6901 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6961 tggccttttg ctggcctttt gctcacatgt
//