LOCUS       sequence-269486-        8876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression human MSH6_T1219D mutant protein using baculovirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8876)
  AUTHORS   Geng H, Du C, Chen S, Salerno V, Manfredi C, Hsieh P
  TITLE     In vitro studies of DNA mismatch repair proteins.
  JOURNAL   Anal Biochem. 2011 Jun 15;413(2):179-84. doi: 
            10.1016/j.ab.2011.02.017. Epub 2011 Feb 15.
  PUBMED    21329650
REFERENCE   2  (bases 1 to 8876)
  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"; doi: 
            "10.1016/j.ab.2011.02.017"; journalName: "Anal Biochem"; date: 
            "2011-06-15- 15"; volume: "413"; issue: "2"; pages: "179-84"
COMMENT     Sequence Label: pFB HTB_hMSH6_T1219D
FEATURES             Location/Qualifiers
     source          1..8876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..92
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     17..34
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             159..176
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             198..218
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    4463..4597
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4500..4519)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4554..4573
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4626..4791
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4975..5429
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5062..5081)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5271..5292
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5455..5559
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5560..5628,5629..6420)
                     /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(5778..5797)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6591..7179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7080..7099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7333..7350
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  7484..7708
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(7775..8308)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(8497..8502,8503..8519,8520..8525))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atctcggtcc gaaaccatgt cgtactacca tcaccatcac catcacgatt
      181 acgatatccc aacgaccgaa aacctgtatt ttcagggcgc catgggatcc atgtcgcgac
      241 agagcaccct gtacagcttc ttccccaagt ctccggcgct gagtgatgcc aacaaggcct
      301 cggccagggc ctcacgcgaa ggcggccgtg ccgccgctgc ccccggggcc tctccttccc
      361 caggcgggga tgcggcctgg agcgaggctg ggcctgggcc caggcccttg gcgcgatccg
      421 cgtcaccgcc caaggcgaag aacctcaacg gagggctgcg gagatcggta gcgcctgctg
      481 cccccaccag ttgtgacttc tcaccgggag atttggtttg ggccaagatg gagggttacc
      541 cctggtggcc ttgtctggtt tacaaccacc cctttgatgg aacattcatc cgcgagaaag
      601 ggaaatcagt ccgtgttcat gtacagtttt ttgatgacag cccaacaagg ggctgggtta
      661 gcaaaaggct tttaaagcca tatacaggtt caaaatcaaa ggaagcccag aagggaggtc
      721 atttttacag tgcaaagcct gaaatactga gagcaatgca acgtgcagac gaagccttaa
      781 ataaagacaa gattaagagg cttgaattgg cagtttgtga tgagccctca gagccagaag
      841 aggaagaaga gatggaggta ggcacaactt acgtaacaga taagagtgaa gaagataatg
      901 aaattgagag tgaagaggaa gtacagccta agacacaagg atctaggcga agtagccgcc
      961 aaataaaaaa acgaagggtc atatcagatt ctgagagtga cattggtggc tctgatgtgg
     1021 aatttaagcc agacactaag gaggaaggaa gcagtgatga aataagcagt ggagtggggg
     1081 atagtgagag tgaaggcctg aacagccctg tcaaagttgc tcgaaagcgg aagagaatgg
     1141 tgactggaaa tggctctctt aaaaggaaaa gctctaggaa ggaaacgccc tcagccacca
     1201 aacaagcaac tagcatttca tcagaaacca agaatacttt gagagctttc tctgcccctc
     1261 aaaattctga atcccaagcc cacgttagtg gaggtggtga tgacagtagt cgccctactg
     1321 tttggtatca tgaaacttta gaatggctta aggaggaaaa gagaagagat gagcacagga
     1381 ggaggcctga tcaccccgat tttgatgcat ctacactcta tgtgcctgag gatttcctca
     1441 attcttgtac tcctgggatg aggaagtggt ggcagattaa gtctcagaac tttgatcttg
     1501 tcatctgtta caaggtgggg aaattttatg agctgtacca catggatgct cttattggag
     1561 tcagtgaact ggggctggta ttcatgaaag gcaactgggc ccattctggc tttcctgaaa
     1621 ttgcatttgg ccgttattca gattccctgg tgcagaaggg ctataaagta gcacgagtgg
     1681 aacagactga gactccagaa atgatggagg cacgatgtag aaagatggca catatatcca
     1741 agtatgatag agtggtgagg agggagatct gtaggatcat taccaagggt acacagactt
     1801 acagtgtgct ggaaggtgat ccctctgaga actacagtaa gtatcttctt agcctcaaag
     1861 aaaaagagga agattcttct ggccatactc gtgcatatgg tgtgtgcttt gttgatactt
     1921 cactgggaaa gtttttcata ggtcagtttt cagatgatcg ccattgttcg agatttagga
     1981 ctctagtggc acactatccc ccagtacaag ttttatttga aaaaggaaat ctctcaaagg
     2041 aaactaaaac aattctaaag agttcattgt cctgttctct tcaggaaggt ctgatacccg
     2101 gctcccagtt ttgggatgca tccaaaactt tgagaactct ccttgaggaa gaatatttta
     2161 gggaaaagct aagtgatggc attggggtga tgttacccca ggtgcttaaa ggtatgactt
     2221 cagagtctga ttccattggg ttgacaccag gagagaaaag tgaattggcc ctctctgctc
     2281 taggtggttg tgtcttctac ctcaaaaaat gccttattga tcaggagctt ttatcaatgg
     2341 ctaattttga agaatatatt cccttggatt ctgacacagt cagcactaca agatctggtg
     2401 ctatcttcac caaagcctat caacgaatgg tgctagatgc agtgacatta aacaacttgg
     2461 agatttttct gaatggaaca aatggttcta ctgaaggaac cctactagag agggttgata
     2521 cttgccatac tccttttggt aagcggctcc taaagcaatg gctttgtgcc ccactctgta
     2581 accattatgc tattaatgat cgtctagatg ccatagaaga cctcatggtt gtgcctgaca
     2641 aaatctccga agttgtagag cttctaaaga agcttccaga tcttgagagg ctactcagta
     2701 aaattcataa tgttgggtct cccctgaaga gtcagaacca cccagacagc agggctataa
     2761 tgtatgaaga aactacatac agcaagaaga agattattga ttttctttct gctctggaag
     2821 gattcaaagt aatgtgtaaa attataggga tcatggaaga agttgctgat ggttttaagt
     2881 ctaaaatcct taagcaggtc atctctctgc agacaaaaaa tcctgaaggt cgttttcctg
     2941 atttgactgt agaattgaac cgatgggata cagcctttga ccatgaaaag gctcgaaaga
     3001 ctggacttat tactcccaaa gcaggctttg actctgatta tgaccaagct cttgctgaca
     3061 taagagaaaa tgaacagagc ctcctggaat acctagagaa acagcgcaac agaattggct
     3121 gtaggaccat agtctattgg gggattggta ggaaccgtta ccagctggaa attcctgaga
     3181 atttcaccac tcgcaatttg ccagaagaat acgagttgaa atctaccaag aagggctgta
     3241 aacgatactg gaccaaaact attgaaaaga agttggctaa tctcataaat gctgaagaac
     3301 ggagggatgt atcattgaag gactgcatgc ggcgactgtt ctataacttt gataaaaatt
     3361 acaaggactg gcagtctgct gtagagtgta tcgcagtgtt ggatgtttta ctgtgcctgg
     3421 ctaactatag tcgagggggt gatggtccta tgtgtcgccc agtaattctg ttgccggaag
     3481 ataccccccc cttcttagag cttaaaggat cacgccatcc ttgcattacg aagacttttt
     3541 ttggagatga ttttattcct aatgacattc taataggctg tgaggaagag gagcaggaaa
     3601 atggcaaagc ctattgtgtg cttgttactg gaccaaatat ggggggcaag tctacgctta
     3661 tgagacaggc tggcttatta gctgtaatgg cccagatggg ttgttacgtc cctgctgaag
     3721 tgtgcaggct cacaccaatt gatagagtgt ttactagact tggtgcctca gacagaataa
     3781 tgtcaggtga aagtacattt tttgttgaat taagtgaaac tgccagcata ctcatgcatg
     3841 caacagcaca ttctctggtg cttgtggatg aattaggaag aggtgatgca acatttgatg
     3901 ggacggcaat agcaaatgca gttgttaaag aacttgctga gactataaaa tgtcgtacat
     3961 tattttcaac tcactaccat tcattagtag aagattattc tcaaaatgtt gctgtgcgcc
     4021 taggacatat ggcatgcatg gtagaaaatg aatgtgaaga ccccagccag gagactatta
     4081 cgttcctcta taaattcatt aagggagctt gtcctaaaag ctatggcttt aatgcagcaa
     4141 ggcttgctaa tctcccagag gaagttattc aaaagggaca tagaaaagca agagaatttg
     4201 agaagatgaa tcagtcacta cgattatttc gggaagtttg cctggctagt gaaaggtcaa
     4261 ctgtagatgc tgaagctgtc cataaattgc tgactttgat taaggaatta tagatctcga
     4321 ggcatgcggt accaagcttg tcgagaagta ctagaggatc ataatcagcc ataccacatt
     4381 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     4441 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     4501 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4561 gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgatat cgcctaggag
     4621 atccgaacca gataagtgaa atctagttcc aaactatttt gtcattttta attttcgtat
     4681 tagcttacga cgctacaccc agttcccatc tattttgtca ctcttcccta aataatcctt
     4741 aaaaactcca tttccacccc tcccagttcc caactatttt gtccgcccac agcggggcat
     4801 ttttcttcct gttatgtttt taatcaaaca tcctgccaac tccatgtgac aaaccgtcat
     4861 cttcggctac tttttctctg tcacagaatg aaaatttttc tgtcatctct tcgttattaa
     4921 tgtttgtaat tgactgaata tcaacgctta tttgcagcct gaatggcgaa tgggacgcgc
     4981 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5041 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5101 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5161 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5221 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttcttaata gtggactctt
     5281 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5341 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5401 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     5461 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5521 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5581 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5641 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5701 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5761 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5821 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5881 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5941 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6001 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6061 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6121 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6181 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6241 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6301 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6361 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6421 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6481 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6541 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6601 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6661 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6721 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6781 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6841 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6901 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6961 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7021 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7081 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7141 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7201 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7261 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7321 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     7381 ctccttacgc atctgtgcgg tatttcacac cgcatagacc agccgcgtaa cctggcaaaa
     7441 tcggttacgg ttgagtaata aatggatgcc ctgcgtaagc gggtgtgggc ggacaataaa
     7501 gtcttaaact gaacaaaata gatctaaact atgacaataa agtcttaaac tagacagaat
     7561 agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag catactggac ttttgttatg
     7621 gctaaagcaa actcttcatt ttctgaagtg caaattgccc gtcgtattaa agaggggcgt
     7681 ggccaagggc atggtaaaga ctatattcgc ggcgttgtga caatttaccg aacaactccg
     7741 cggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatat
     7801 caaagtgcat cacttcttcc cgtatgccca actttgtata gagagccact gcgggatcgt
     7861 caccgtaatc tgcttgcacg tagatcacat aagcaccaag cgcgttggcc tcatgcttga
     7921 ggagattgat gagcgcggtg gcaatgccct gcctccggtg ctcgccggag actgcgagat
     7981 catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa gccgcgagag
     8041 cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta ctacggagca
     8101 agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg tctccgaact
     8161 cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg ccgagcctac
     8221 atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga ctgccctgct
     8281 gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc
     8341 acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca
     8401 taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg
     8461 gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat
     8521 gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc
     8581 agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc
     8641 acgcatcgtc aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat
     8701 ctgccctggc ttcaggagat cggaagacct cggccgtcgc ggcgcttgcc ggtggtgctg
     8761 accccggatg aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc
     8821 caggactcta gctatagttc tagtggttgg ctacgtatac tccggaatat taatag
//
