LOCUS       Exported                6620 bp ds-DNA     circular SYN 07-AUG-2020
DEFINITION  His6-MBP-tev-G-Hs.KRAS(1-188).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6620)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Aug 12, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6620
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..92
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             137..1894
                     /codon_start=1
                     /gene="tev-G-Hs.KRAS(1-188) G12C"
                     /label=H6-MBP-tev-G-Hs.KRAS4b(1-188)
                     /translation="MGSHHHHHHGKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKV
                     TVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTW
                     DAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQE
                     PYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSI
                     AEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASP
                     NKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIM
                     PNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSGSSITSLYKKVGENLYFQGMT
                     EYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVIDGETCLLDILDTAGQ
                     EEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVKDSEDVPMVLVGNKCDL
                     PSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREIRKHKEKMSKDGKKKKK
                     KSKTKCVIM"
     CDS             146..163
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             167..1264
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1121..1130
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    1283..1307
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1307..1894
                     /codon_start=1
                     /label=tev-G-Hs.KRAS4b(1-188)
                     /translation="ENLYFQGMTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIED
                     SYRKQVVIDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYR
                     EQIKRVKDSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAF
                     YTLVREIRKHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             1307..1327
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(1893..1917)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2062..2196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  2225..2390
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2574..3029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3056..3160
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3161..4021
                     /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      4192..4780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     mobile_element  5083..5307
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5374..5907)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(6096..6124)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(6488..6597)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
ORIGIN
        1 atcatggaga taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc
       61 gtaacagttt tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca
      121 tcgggcgcgg atcaccatgg gatcccatca ccatcaccat cacggcaaaa tcgaagaagg
      181 taaactggta atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa
      241 gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata aactggaaga
      301 gaaattccca caggttgcgg caactggcga tggccctgac attatcttct gggcacacga
      361 ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt
      421 ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc
      481 ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc
      541 gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag
      601 cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg
      661 gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa
      721 cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa
      781 tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac
      841 catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac
      901 ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg
      961 tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct
     1021 gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa
     1081 gtcttacgag gaagagttgg cgaaagatcc acgtattgcc gccaccatgg aaaacgccca
     1141 gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac
     1201 tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca
     1261 gactaattcg ggatcctcga tcacaagttt gtacaaaaaa gttggcgaaa acctgtactt
     1321 ccaaggcatg actgaatata aacttgtggt agttggagct ggtggcgtag gcaagagtgc
     1381 cttgacgata cagctaattc agaatcattt tgtggacgaa tatgatccaa caatagagga
     1441 ttcctacagg aagcaagtag taattgatgg agaaacctgt ctcttggata ttctcgacac
     1501 agcaggtcaa gaggagtaca gtgcaatgag ggaccagtac atgaggactg gggagggctt
     1561 tctttgtgta tttgccataa ataatactaa atcatttgaa gatattcacc attatagaga
     1621 acaaattaaa agagttaagg actctgaaga tgtacctatg gtcctagtag gaaataaatg
     1681 tgatttgcct tctagaacag tagacacaaa acaggctcag gacttagcaa gaagttatgg
     1741 aattcctttt attgaaacat cagcaaagac aagacagggt gttgatgatg ccttctatac
     1801 attagttcga gaaattcgaa aacataaaga aaagatgagc aaagatggta aaaagaagaa
     1861 aaagaagtca aagacaaagt gtgtaattat gtaatcaact ttcttgtaca aagtggtgat
     1921 cgagctagct agaagcttgt cgagaagtac tagaggatca taatcagcca taccacattt
     1981 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2041 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2101 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2161 tccaaactca tcaatgtatc ttatcatgtc tggatctgat cactgcttga gcctaggaga
     2221 tccgaaccag ataagtgaaa tctagttcca aactattttg tcatttttaa ttttcgtatt
     2281 agcttacgac gctacaccca gttcccatct attttgtcac tcttccctaa ataatcctta
     2341 aaaactccat ttccacccct cccagttccc aactattttg tccgcccaca gcggggcatt
     2401 tttcttcctg ttatgttttt aatcaaacat cctgccaact ccatgtgaca aaccgtcatc
     2461 ttcggctact ttttctctgt cacagaatga aaatttttct gtcatctctt cgttattaat
     2521 gtttgtaatt gactgaatat caacgcttat ttgcagcctg aatggcgaat gggacgcgcc
     2581 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2641 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2701 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2761 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2821 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2881 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2941 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3001 ttttaacaaa atattaacgt ttacaatttc aggtggcact tttcggggaa atgtgcgcgg
     3061 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3121 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3181 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3241 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3301 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3361 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3421 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3481 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3541 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3601 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3661 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3721 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3781 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3841 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3901 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3961 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4021 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4081 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4141 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4201 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4261 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4321 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4381 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4441 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4501 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4561 actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc
     4621 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4681 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4741 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4801 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4861 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4921 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     4981 tctccttacg catctgtgcg gtatttcaca ccgcagacca gccgcgtaac ctggcaaaat
     5041 cggttacggt tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag
     5101 tcttaaactg aacaaaatag atctaaacta tgacaataaa gtcttaaact agacagaata
     5161 gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc atactggact tttgttatgg
     5221 ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg
     5281 gccaagggca tggtaaagac tatattcgcg gcgttgtgac aatttaccga acaactccgc
     5341 ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc
     5401 aaagtgcatc acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc
     5461 accgtaatct gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag
     5521 gagattgatg agcgcggtgg caatgccctg cctccggtgc tcgccggaga ctgcgagatc
     5581 atagatatag atctcactac gcggctgctc aaacctgggc agaacgtaag ccgcgagagc
     5641 gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa
     5701 gttcccgagg taatcggagt ccggctgatg ttgggagtag gtggctacgt ctccgaactc
     5761 acgaccgaaa agatcaagag cagcccgcat ggatttgact tggtcagggc cgagcctaca
     5821 tgtgcgaatg atgcccatac ttgagccacc taactttgtt ttagggcgac tgccctgctg
     5881 cgtaacatcg ttgctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca
     5941 cggcgtaacg cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa aaacagtcat
     6001 aacaagccat gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg
     6061 accagttgcg tgagcgcata cgctacttgc attacagttt acgaaccgaa caggcttatg
     6121 tcaactgggt tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggca
     6181 gcagcgaagt cgaggcattt ctgtcctggc tggcgaacga gcgcaaggtt tcggtctcca
     6241 cgcatcgtca ggcattggcg gccttgctgt tcttctacgg caaggtgctg tgcacggatc
     6301 tgccctggct tcaggagatc ggtagacctc ggccgtcgcg gcgcttgccg gtggtgctga
     6361 ccccggatga agtggttcgc atcctcggtt ttctggaagg cgagcatcgt ttgttcgccc
     6421 aggactctag ctatagttct agtggttggc ctactcgaga tcccgggtga tcaagtcttc
     6481 gtcgagtgat tgtaaataaa atgtaattta cagtatagta ttttaattaa tatacaaatg
     6541 atttgataat aattcttatt taactataat atattgtgtt gggttgaatt aaaggtccgt
     6601 atactccgga atattaatag
//
