LOCUS       sequence-299863-        5608 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses human Kras wild-type.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5608)
  TITLE     Mutant Ras Constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5608)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pBabe-Kras Wt
FEATURES             Location/Qualifiers
     source          1..5608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    467..666
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             867..1283
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1213..1235
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1251..1267
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1306..1332
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1312..1338
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             1360..1926
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     primer_bind     complement(1938..1958)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1943..2272
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2123..2258
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2282..2881
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2282..2301)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2778..2798
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3102..3571
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3701..3718)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3872..4460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3952..3971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4620..5411,5412..5480))
                     /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     5243..5262
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5481..5585)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat
      121 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
      181 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
      241 ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
      301 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
      361 tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg ctcgtccggg
      421 atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggcaagc tggccagcaa
      481 cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc ctgcgtcggt
      541 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      601 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      661 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      721 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      781 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      841 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
      901 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
      961 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1021 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1081 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1141 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1201 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1261 ccctcactcc ttctctaggc gccggccgga tcaatggctt caagttatcc atatgatgtg
     1321 cccgactatg cctcacttgg aggaccctct ggatccacca tgactgaata taaacttgtg
     1381 gtagttggag ctggcggcgt aggcaagagt gccttgacga tacagctaat tcagaatcat
     1441 tttgtggacg aatatgatcc aacaatagag gattcctaca ggaagcaagt agtaattgat
     1501 ggagaaacct gtctcttgga tattctcgac acagcaggtc aagaggagta cagtgcaatg
     1561 agggaccagt acatgaggac tggggagggc tttctttgtg tatttgccat aaataatact
     1621 aaatcatttg aagatattca ccattataga gaacaaatta aaagagttaa ggactctgaa
     1681 gatgtaccta tggtcctagt aggaaataaa tgtgatttgc cttccagaac agtagacaca
     1741 aaacaggctc aggacttagc aagaagttat ggaattcctt ttattgaaac atcagcaaag
     1801 acaagacagg gtgttgatga tgccttctat acattagttc gagaaattcg aaaacataaa
     1861 gaaaagatga gcaaagatgg taaaaagaag aaaaagaagt caaagacaaa gtgtgtaatt
     1921 atgtaagtcg accctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     1981 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2041 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2101 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2161 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     2221 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac
     2281 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     2341 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     2401 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     2461 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     2521 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     2581 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     2641 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     2701 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     2761 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     2821 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     2881 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcgat
     2941 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     3001 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag
     3061 aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag
     3121 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     3181 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     3241 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     3301 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     3361 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     3421 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     3481 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     3541 tgattgacta cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt
     3601 tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg
     3661 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3721 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3781 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3841 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3901 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3961 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4021 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4081 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4141 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4201 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4261 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4321 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4381 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4441 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4501 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4561 agatcctttt gcggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4621 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4681 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4741 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4801 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4861 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4921 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4981 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5041 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5101 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5161 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5221 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5281 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5341 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5401 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5461 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5521 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5581 ccgcgcacat ttccccgaaa agtgccac
//
