LOCUS       sequence-486868-        2007 bp    DNA     circular SYN 22-SEP-2025
DEFINITION  Express SmBiT-KRAS(G12D) Fusion Protein in Mammalian Cells under a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2007)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2007)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: SmBiT-KRAS(G12D) BiBiT-Ready Vector
FEATURES             Location/Qualifiers
     source          1..2007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(183..386)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(212..232)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        387..690
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    923..1044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(960..979)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1014..1033
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(1190..1720,1721..1723,1724..1756))
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase K-Ras"
                     /label=K-Ras (G12V)
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             complement(1190..1756)
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     primer_bind     complement(1859..1878)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1860..1878)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1920..1939)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          join(1922..2007,1..47)
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
ORIGIN
        1 agtttctatt ggtctcctta aacctgtctt gtaaccttga tacttacctg cccagtgcct
       61 cacgaccaac ttctgcagct taagttcgag actgttgtgt cagaagcact gactgcgtta
      121 gcaatttaac tgtgataaac taccgcaata aagcttctag tgatctgacg gttcactaaa
      181 ccagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaacggg
      241 gcggggttat tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
      301 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
      361 ttggtgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
      421 tgccaagtag gaaagtcccg taaggtcatg tactgggcat aatgccaggc gggccattta
      481 ccgtcattga cgtcaatagg gggcggactt ggcatatgat acacttgatg tactgccaag
      541 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
      601 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
      661 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
      721 gaccccgtaa ttgattacta ttaataacta gtggcgcgcc ataatcctag gctctcgtac
      781 gaagtcagct gataacgtcg acgtattgcg gccgcggatc ataatcagcc ataccacatt
      841 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
      901 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
      961 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1021 gtccaaactc atcaatgtat cttatcatgt ctgctcgaag cggccttgct cagcggtggc
     1081 agcagccaac tcagcttcct ttcgggcttt gttagcagcc ggatcagctt gcatgcctgc
     1141 aggtcgactc tagaggatcc ccgggtaccg agcccgaatt cgtttaaact tacataatta
     1201 cacactttgt ctttgacttc tttttcttct ttttaccatc tttgctcatc ttttctttat
     1261 gttttcgaat ttctcgaact aatgtataga aggcatcatc aacaccctgt cttgtctttg
     1321 ctgatgtttc aataaaagga attccataac ttcttgctaa gtcctgagcc tgttttgtgt
     1381 ctactgttct agaaggcaaa tcacatttat ttcctactag gaccataggt acatcttcag
     1441 agtccttaac tcttttaatt tgttctctat aatggtgaat atcttcaaat gatttagtat
     1501 tatttatggc aaatacacaa agaaagccct ccccagtcct catgtactgg tccctcattg
     1561 cactgtactc ctcttgacct gctgtgtcga gaatatccaa gagacaggtt tctccatcaa
     1621 ttactacttg cttcctgtag gaatcctcta ttgttggatc atattcgtcc acaaaatgat
     1681 tctgaattag ctgtatcgtc aaggcactct tgcctacgcc gtcagctcca actaccacaa
     1741 gtttatattc agtcatggcg atcgcaccgc tcgagcctcc acctccgctc ccgccaccac
     1801 cggagctccc gagaatctcc tcgaacagcc ggtagccggt caccatggtg agcgctagcc
     1861 ctatagtgag tcgtattaat actctagcct taagagctgt aattgaactg ggagtggaca
     1921 cctgtggaga gaaaggcaaa gtggatgtca gtaagaccaa taggtgccta tcagaaacgc
     1981 aagagtcttc tctgtctcga caagccc
//
