LOCUS       sequence-472710-        5868 bp    DNA     circular SYN 02-JUL-2025
DEFINITION  Express HiBiT-KRAS(G12C) in Mammalian Cells under a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5868)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5868)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: HiBiT-KRAS(G12C) Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1134..1700
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGACGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     primer_bind     complement(1814..1832)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    1869..1990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1880..1899)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1934..1953
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2235..2592
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2235..2426
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2258..2278)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2443..2578
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2643..3437
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2697..2716)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3307..3326
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3501..3549
                     /note="synthetic polyadenylation signal"
     promoter        3705..3809
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3810..3878,3879..4670)
                     /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(4028..4047)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4828..5416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4908..4927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5532..5815
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatgactg
     1141 aatataaact tgtggtagtt ggagcttgcg gcgtaggcaa gagtgccttg acgatacagc
     1201 taattcagaa tcattttgtg gacgaatatg atccaacaat agaggattcc tacaggaagc
     1261 aagtagtaat tgatggagaa acctgtctct tggatattct cgacacagca ggtcaagagg
     1321 agtacagtgc aatgagggac cagtacatga ggactgggga gggctttctt tgtgtatttg
     1381 ccataaataa tactaaatca tttgaagata ttcaccatta tagagaacaa attaaaagag
     1441 ttaaggactc tgaagatgta cctatggtcc tagtaggaaa taaatgtgat ttgccttcta
     1501 gaacagtaga cacaaaacag gctcaggact tagcaagaag ttatggaatt ccttttattg
     1561 aaacatcagc aaagacaaga cagggtgttg atgatgcctt ctatacatta gttcgagaaa
     1621 ttcgaaaaca taaagaaaag atgagcaaag atggtaaaaa gaagaaaaag aagtcaaaga
     1681 caaagtgtgt aattatgtaa gtttaaacga attcgggctc ggtacccggg gatcctctag
     1741 agtcgacctg caggcatgca agctgatccg gctgctaaca aagcccgaaa ggaagctgag
     1801 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcggc cgcttcgagc
     1861 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1921 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1981 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     2041 ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa
     2101 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2161 acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
     2221 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     2281 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2341 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2401 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2461 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2521 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2581 ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag gctagagcca
     2641 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2701 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2761 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2821 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2881 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2941 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3001 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3061 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3121 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     3181 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3241 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3301 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3361 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3421 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     3481 cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     3541 ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga
     3601 tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg
     3661 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3721 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3781 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3841 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3901 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3961 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttca
     4021 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4081 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4141 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4201 ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc gcttttttgc
     4261 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4321 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4381 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4441 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4501 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4561 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4621 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ttcgaaatga
     4681 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4741 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4801 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4861 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4921 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4981 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5041 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5101 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5161 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5221 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5281 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5341 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5401 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5461 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5521 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     5581 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     5641 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     5701 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     5761 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     5821 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
