LOCUS Exported 7906 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pQCXP CMV/TO RasV12 (w364-1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7906) TITLE shMDC1 plasmids REFERENCE 2 (bases 1 to 7906) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7906 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 309..512 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 513..688 /label=5' LTR (truncated) /note="truncated long terminal repeat from Moloney murine sarcoma virus" misc_feature 751..950 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1151..1567 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1497..1519 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1535..1551 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" enhancer 1609..1912 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1913..2116 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 2118..2136 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2139..2157 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2198..2222 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2297..2866 /codon_start=1 /product="human oncoprotein generated by the G12V mutation in the small GTPase H-Ras" /label=H-Ras (G12V) /translation="MTEYKLVVVGAVGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI RQHKLRKLNPPDESGPGCMSCKCVLS" protein_bind complement(3029..3053) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" promoter 3119..3618 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(3154..3177) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 3551..3570 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 3639..4238 /codon_start=1 /gene="pac from Streptomyces" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA" primer_bind complement(3639..3658) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4135..4155 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 4612..5037 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus" primer_bind complement(5140..5162) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5298..5318) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5303..5632 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5483..5618 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5545..5564 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 5654..5673 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(5754..5771) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5925..6513) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6005..6024) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6684..7544) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7307..7326 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7545..7649) /gene="bla" /label=AmpR promoter primer_bind 7717..7735 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" enhancer 7835..308 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc 61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac 121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata 181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc 241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta 301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac 361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc 421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc 481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc 541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc 601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat 661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc 721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc 781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta 841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc 901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc 961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca 1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct 1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc 1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga 1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt 1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg 1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga 1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa 1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct 1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct 1561 aggcgccgga attgaagatc tgggggatcg ataagcttgg gagttccgcg ttacataact 1621 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat 1681 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta 1741 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc 1801 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg 1861 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg 1921 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct 1981 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa 2041 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt 2101 ctatataagc agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg 2161 tcgactagtc cagtgtggtg gaattctgca gatatcaaca agtttgtaca aaaaagcagg 2221 ctttaaagga accaattcag tcgactggat cccagtgtgg tggtacagct tggggggcca 2281 gggccggggc cgagcgatga cggaatataa gctggtggtg gtgggcgccg tcggtgtggg 2341 caagagtgcg ctgaccatcc agctgatcca gaaccatttt gtggacgaat acgaccccac 2401 tatagaggat tcctaccgga agcaggtggt cattgatggg gagacgtgcc tgttggacat 2461 cctggatacc gccggccagg aggagtacag cgccatgcgg gaccagtaca tgcgcaccgg 2521 ggagggcttc ctgtgtgtgt ttgccatcaa caacaccaag tcttttgagg acatccacca 2581 gtacagggag cagatcaaac gggtgaagga ctcggatgac gtgcccatgg tgctggtggg 2641 gaacaagtgt gacctggctg cacgcactgt ggaatctcgg caggctcagg acctcgcccg 2701 aagctacggc atcccctaca tcgagacctc ggccaagacc cggcagggag tggaggatgc 2761 cttctacacg ttggtgcgtg agatccggca gcacaagctg cggaagctga accctcctga 2821 tgagagtggc cccggctgca tgagctgcaa gtgtgtgctc tcctgacgca gcacaagctc 2881 aggacatgga ggtgccggat gcaggaagga ggtgcagacg gaaggaggag gaaggaagga 2941 cggaagcaag gaaggaagga agggctgctg gagcccagtc accccccaag ctgtaggaat 3001 tcgccagcac agtggtcgag atatctagac ccagctttct tgtacaaagt ggttgatatc 3061 cagcacagtg gcgaccggta ggcctcgtac gcttaattaa cggatccgga attctaccgg 3121 gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc 3181 acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc 3241 caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg 3301 aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt 3361 ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg 3421 ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt 3481 gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc 3541 ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc 3601 tcatctccgg gcctttcgac ctgcagccca agcttaccat gaccgagtac aagcccacgg 3661 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg 3721 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg 3781 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg 3841 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt 3901 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac 3961 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg 4021 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag 4081 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc 4141 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgagtgc ccgaaggacc 4201 gcgcgacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac gacccgcagc 4261 gcccgaccga aaggagcgca cgaccccatg gctccgaccg aagccgaccc gggcggcccc 4321 gccgaccccg cacccgcccc cgaggcccac cgactctagt cgagggctgc agcgctgcag 4381 aggccgagtg cagaactgct ccaaagggac ctcaaggctt tccgagggac actaggctga 4441 ctccatcgag ccagtgtaga gataagctta tcgattagtc caatttgtta aagacaggat 4501 atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta 4561 cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga 4621 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa 4681 tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa cagggtcgac 4741 cctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt 4801 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc 4861 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc 4921 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg 4981 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg 5041 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta 5101 agctggctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc 5161 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc 5221 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg 5281 agtgtagatc cggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc 5341 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc 5401 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat 5461 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc 5521 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga 5581 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac 5641 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa 5701 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc 5761 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 5821 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 5881 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 5941 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 6001 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 6061 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 6121 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 6181 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 6241 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 6301 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 6361 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 6421 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 6481 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 6541 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 6601 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 6661 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 6721 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 6781 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 6841 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 6901 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 6961 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc 7021 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 7081 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 7141 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 7201 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 7261 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca 7321 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 7381 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 7441 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 7501 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 7561 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 7621 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 7681 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt 7741 cgtcttcaag aattagcttg gccattgcat acgttgtatc catatcataa tatgtacatt 7801 tatattggct catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa 7861 tagtaatcaa ttacggggtc attagttcat agcccatata tggagt //