LOCUS       sequence-254831-        6966 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression human MLH1 protein using baculovirus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6966)
  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 6966)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pFB1_hMLH1
FEATURES             Location/Qualifiers
     source          1..6966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  1..225
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(292..825)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1014..1019,1020..1036,1037..1042))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        1394..1485
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1410..1427
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    3946..4080
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3983..4002)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4037..4056
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4109..4274
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4458..4912
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4545..4564)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4754..4775
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4938..5042
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5043..5111,5112..5903)
                     /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(5261..5280)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6074..6662
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6563..6582
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6816..6833
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgtgggcgga caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt
       61 cttaaactag acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat
      121 actggacttt tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc
      181 gtattaaaga ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa
      241 tttaccgaac aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc
      301 ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag
      361 agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc
      421 gttggcctca tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc
      481 gccggagact gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag
      541 aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa
      601 tgtcttacta cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt
      661 ggctacgtct ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg
      721 gtcagggccg agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt
      781 agggcgactg ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata
      841 acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact
      901 gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc
      961 gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac
     1021 gaaccgaaca ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc
     1081 acccggcaac cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc
     1141 gcaaggtttc ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca
     1201 aggtgctgtg cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc
     1261 gcttgccggt ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg
     1321 agcatcgttt gttcgcccag gactctagct atagttctag tggttggcta cgtatactcc
     1381 ggaatattaa tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta
     1441 ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca
     1501 taccgtccca ccatcgggcg cggatccatg tcgttcgtgg caggggttat tcggcggctg
     1561 gacgagacag tggtgaaccg catcgcggcg ggggaagtta tccagcggcc agctaatgct
     1621 atcaaagaga tgattgagaa ctgtttagat gcaaaatcca caagtattca agtgattgtt
     1681 aaagagggag gcctgaagtt gattcagatc caagacaatg gcaccgggat caggaaagaa
     1741 gatctggata ttgtatgtga aaggttcact actagtaaac tgcagtcctt tgaggattta
     1801 gccagtattt ctacctatgg ctttcgaggt gaggctttgg ccagcataag ccatgtggct
     1861 catgttacta ttacaacgaa aacagctgat ggaaagtgtg catacagagc aagttactca
     1921 gatggaaaac tgaaagcccc tcctaaacca tgtgctggca atcaagggac ccagatcacg
     1981 gtggaggacc ttttttacaa catagccacg aggagaaaag ctttaaaaaa tccaagtgaa
     2041 gaatatggga aaattttgga agttgttggc aggtattcag tacacaatgc aggcattagt
     2101 ttctcagtta aaaaacaagg agagacagta gctgatgtta ggacactacc caatgcctca
     2161 accgtggaca atattcgctc catctttgga aatgctgtta gtcgagaact gatagaaatt
     2221 ggatgtgagg ataaaaccct agccttcaaa atgaatggtt acatatccaa tgcaaactac
     2281 tcagtgaaga agtgcatctt cttactcttc atcaaccatc gtctggtaga atcaacttcc
     2341 ttgagaaaag ccatagaaac agtgtatgca gcctatttgc ccaaaaacac acacccattc
     2401 ctgtacctca gtttagaaat cagtccccag aatgtggatg ttaatgtgca ccccacaaag
     2461 catgaagttc acttcctgca cgaggagagc atcctggagc gggtgcagca gcacatcgag
     2521 agcaagctcc tgggctccaa ttcctccagg atgtacttca cccagacttt gctaccagga
     2581 cttgctggcc cctctgggga gatggttaaa tccacaacaa gtctgacctc gtcttctact
     2641 tctggaagta gtgataaggt ctatgcccac cagatggttc gtacagattc ccgggaacag
     2701 aagcttgatg catttctgca gcctctgagc aaacccctgt ccagtcagcc ccaggccatt
     2761 gtcacagagg ataagacaga tatttctagt ggcagggcta ggcagcaaga tgaggagatg
     2821 cttgaactcc cagcccctgc tgaagtggct gccaaaaatc agagcttgga gggggataca
     2881 acaaagggga cttcagaaat gtcagagaag agaggaccta cttccagcaa ccccagaaag
     2941 agacatcggg aagattctga tgtggaaatg gtggaagatg attcccgaaa ggaaatgact
     3001 gcagcttgta ccccccggag aaggatcatt aacctcacta gtgttttgag tctccaggaa
     3061 gaaattaatg agcagggaca tgaggttctc cgggagatgt tgcataacca ctccttcgtg
     3121 ggctgtgtga atcctcagtg ggccttggca cagcatcaaa ccaagttata ccttctcaac
     3181 accaccaagc ttagtgaaga actgttctac cagatactca tttatgattt tgccaatttt
     3241 ggtgttctca ggttatcgga gccagcaccg ctctttgacc ttgccatgct tgccttagat
     3301 agtccagaga gtggctggac agaggaagat ggtcccaaag aaggacttgc tgaatacatt
     3361 gttgagtttc tgaagaagaa ggctgagatg cttgcagact atttctcttt ggaaattgat
     3421 gaggaaggga acctgattgg attacccctt ctgattgaca actatgtgcc ccctttggag
     3481 ggactgccta tcttcattct tcgactagcc actgaggtga attgggacga agaaaaggaa
     3541 tgttttgaaa gcctcagtaa agaatgcgct atgttctatt ccatccggaa gcagtacata
     3601 tctgaggagt cgaccctctc aggccagcag agtgaagtgc ctggctccat tccaaactcc
     3661 tggaagtgga ctgtggaaca cattgtctat aaagccttgc gctcacacat tctgcctcct
     3721 aaacatttca cagaagatgg aaatatcctg cagcttgcta acctgcctga tctatacaaa
     3781 gtctttgaga ggtgttaact cgaggcatgc ggtaccaagc ttgtcgagaa gtactagagg
     3841 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3901 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3961 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4021 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     4081 tgatcactgc ttgagcctag gagatccgaa ccagataagt gaaatctagt tccaaactat
     4141 tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg
     4201 tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat
     4261 tttgtccgcc cacagcgggg catttttctt cctgttatgt ttttaatcaa acatcctgcc
     4321 aactccatgt gacaaaccgt catcttcggc tactttttct ctgtcacaga atgaaaattt
     4381 ttctgtcatc tcttcgttat taatgtttgt aattgactga atatcaacgc ttatttgcag
     4441 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4501 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4561 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4621 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4681 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4741 cacgttctta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4801 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4861 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     4921 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4981 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5041 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5101 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5161 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5221 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5281 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5341 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5401 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5461 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5521 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5581 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5641 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5701 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5761 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5821 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5881 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5941 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6001 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6061 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6121 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6181 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6241 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6301 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6361 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6421 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6481 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6541 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6601 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6661 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6721 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6781 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6841 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
     6901 accagccgcg taacctggca aaatcggtta cggttgagta ataaatggat gccctgcgta
     6961 agcggg
//
