LOCUS       sequence-313431-        5298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus protein expression of KRAS4b (1-188).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5298)
  AUTHORS   Dharmaiah S, Tran TH, Messing S, Agamasu C, Gillette WK, Yan W, 
            Waybright T, Alexander P, Esposito D, Nissley DV, McCormick F, 
            Stephen AG, Simanshu DK
  TITLE     Structures of N-terminally processed KRAS provide insight into the 
            role of N-acetylation.
  JOURNAL   Sci Rep. 2019 Jul 19;9(1):10512. doi: 10.1038/s41598-019-46846-w.
  PUBMED    31324887
REFERENCE   2  (bases 1 to 5298)
  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"; journalName: "Sci Rep."; 
            date: "2019-07-19"; volume: "9(1):10512. doi"; pages: " 
            10.1038/s41598-019-46846-w"
COMMENT     Sequence Label: R979-X58-008: Hs.KRAS4b(1-188)
FEATURES             Location/Qualifiers
     source          1..5298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..115
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     40..57
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    160..184
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             187..753
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     protein_bind    complement(752..776)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    908..1042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(945..964)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     999..1018
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1071..1236
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1420..1874
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1507..1526)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1716..1737
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1900..2004
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2005..2073,2074..2865)
                     /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(2223..2242)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3036..3624
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3525..3544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3778..3795
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3929..4153
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4220..4753)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4942..4947,4948..4964,4965..4970))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 cgtatactcc ggaatattaa tagatcatgg agataattaa aatgataacc atctcgcaaa
       61 taaataagta ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta taaatattcc
      121 ggattattca taccgtccca ccatcgggcg cggatcatca caagtttgta caaaaaagtt
      181 ggcaccatga ctgaatataa acttgtggta gttggagctg gtggcgtagg caagagtgcc
      241 ttgacgatac agctaattca gaatcatttt gtggacgaat atgatccaac aatagaggat
      301 tcctacagga agcaagtagt aattgatgga gaaacctgtc tcttggatat tctcgacaca
      361 gcaggtcaag aggagtacag tgcaatgagg gaccagtaca tgaggactgg ggagggcttt
      421 ctttgtgtat ttgccataaa taatactaaa tcatttgaag atattcacca ttatagagaa
      481 caaattaaaa gagttaagga ctctgaagat gtacctatgg tcctagtagg aaataaatgt
      541 gatttgcctt ctagaacagt agacacaaaa caggctcagg acttagcaag aagttatgga
      601 attcctttta ttgaaacatc agcaaagaca agacagggtg ttgatgatgc cttctataca
      661 ttagttcgag aaattcgaaa acataaagaa aagatgagca aagatggtaa aaagaagaaa
      721 aagaagtcaa agacaaagtg tgtaattatg taatcaactt tcttgtacaa agtggtgata
      781 gcttgtcgag aagtactaga ggatcataat cagccatacc acatttgtag aggttttact
      841 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
      901 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
      961 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1021 tgtatcttat catgtctgga tctgatcact gcttgagcct aggagatccg aaccagataa
     1081 gtgaaatcta gttccaaact attttgtcat ttttaatttt cgtattagct tacgacgcta
     1141 cacccagttc ccatctattt tgtcactctt ccctaaataa tccttaaaaa ctccatttcc
     1201 acccctccca gttcccaact attttgtccg cccacagcgg ggcatttttc ttcctgttat
     1261 gtttttaatc aaacatcctg ccaactccat gtgacaaacc gtcatcttcg gctacttttt
     1321 ctctgtcaca gaatgaaaat ttttctgtca tctcttcgtt attaatgttt gtaattgact
     1381 gaatatcaac gcttatttgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     1441 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1501 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1561 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     1621 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     1681 ttcgcccttt gacgttggag tccacgttct taatagtgga ctcttgttcc aaactggaac
     1741 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     1801 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     1861 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1921 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1981 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2041 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2101 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2161 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2221 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2281 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2341 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2401 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2461 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2521 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2581 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2641 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2701 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2761 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2821 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2881 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     2941 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3001 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3061 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3121 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3181 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3241 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3301 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3361 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3421 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3481 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3541 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3601 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3661 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3721 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3781 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     3841 tgcggtattt cacaccgcat agaccagccg cgtaacctgg caaaatcggt tacggttgag
     3901 taataaatgg atgccctgcg taagcgggtg tgggcggaca ataaagtctt aaactgaaca
     3961 aaatagatct aaactatgac aataaagtct taaactagac agaatagttg taaactgaaa
     4021 tcagtccagt tatgctgtga aaaagcatac tggacttttg ttatggctaa agcaaactct
     4081 tcattttctg aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca agggcatggt
     4141 aaagactata ttcgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga
     4201 tctcggcttg aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt
     4261 cttcccgtat gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt
     4321 gcacgtagat cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg
     4381 cggtggcaat gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct
     4441 cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt
     4501 cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat
     4561 cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat
     4621 caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc
     4681 ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
     4741 tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct
     4801 tgctgcttgg atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa
     4861 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
     4921 cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggttcgt
     4981 gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag
     5041 gcatttctgt cctggctggc gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca
     5101 ttggcggcct tgctgttctt ctacggcaag gtgctgtgca cggatctgcc ctggcttcag
     5161 gagatcggaa gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc ggatgaagtg
     5221 gttcgcatcc tcggttttct ggaaggcgag catcgtttgt tcgcccagga ctctagctat
     5281 agttctagtg gttggcta
//
