LOCUS Exported 6878 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION gateway adapted mammalian expression vector. ACCESSION . VERSION . KEYWORDS pBABEpuro-gateway SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6878) AUTHORS Greulich H, Kaplan B, Mertins P, Chen TH, Tanaka KE, Yun CH, Zhang X, Lee SH, Cho J, Ambrogio L, Liao R, Imielinski M, Banerji S, Berger AH, Lawrence MS, Zhang J, Pho NH, Walker SR, Winckler W, Getz G, Frank D, Hahn WC, Eck MJ, Mani DR, Jaffe JD, Carr SA, Wong KK, Meyerson M TITLE Functional analysis of receptor tyrosine kinase mutations in lung cancer identifies oncogenic extracellular domain mutations of ERBB2. JOURNAL Proc Natl Acad Sci U S A. 2012 Sep 4;109(36):14476-81. Epub 2012 Aug 20. PUBMED 22908275 REFERENCE 2 (bases 1 to 6878) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6878 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 8..477 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 542..741 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 934..1350 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1318..1334 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" protein_bind 1378..1502 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 1527..1557 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 1611..2270 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1677..1696) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 2612..2917 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 2796..2815 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(2958..3082) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(3118..3138) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3123..3452 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3303..3438 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3365..3384 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3462..4061 /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(3462..3481) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3958..3978 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 4160..4753 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(4954..4971) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5125..5712) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5205..5224) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5890..6750) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6513..6532 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6751..6855) /gene="bla" /label=AmpR promoter ORIGIN 1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca 61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt 121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag 181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc 241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga 301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc 361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc 421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt 481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg 541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta 601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa 661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc 721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg 781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt 841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt 901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt 961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc 1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg 1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca 1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct 1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt 1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt 1321 tatccagccc tcactccttc tctaggcgcc ggccggatcc cagtgtggtg gtacatcaca 1381 agtttgtaca aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat 1441 tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta 1501 tggcggccgc attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt 1561 ggattttgag ttaggatccg tcgagatttt caggagctaa ggaagctaaa atggagaaaa 1621 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg 1681 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct 1741 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg 1801 cccgcctgat gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga 1861 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat 1921 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg 1981 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt 2041 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg 2101 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc 2161 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa 2221 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa acgcgtggat 2281 ccggcttact aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa 2341 gaatatatac tgatatgtat acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg 2401 tattacagtg acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat 2461 atctccggtc tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc 2521 tggaaagcgg aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc 2581 tcttttgctg acgagaacag gggctggtga aatgcagttt aaggtttaca cctataaaag 2641 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg 2701 acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact 2761 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag 2821 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat 2881 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag 2941 ccagtctgca ggtcgaccat agtgactgga tatgttgtgt tttacagtat tatgtagtct 3001 gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt 3061 cagctttctt gtacaaagtg gtgatgtagg aattcgccag cacagtggtc gacctgtgga 3121 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3181 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 3241 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 3301 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 3361 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag 3421 gaggcttttt tggaggccta ggcttttgca aaaagcttac catgaccgag tacaagccca 3481 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt 3541 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca 3601 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg 3661 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg 3721 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc 3781 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca 3841 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg 3901 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca 3961 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag tgcccgaagg 4021 accgcgcgac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc 4081 agcgcccgac cgaaaggagc gcacgacccc atgcatcgat aaaataaaag attttattta 4141 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag 4201 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa 4261 ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt 4321 cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat 4381 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt 4441 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga 4501 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg 4561 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc 4621 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg 4681 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg 4741 ggggtctttc atttccgact tgtggtctcg ctgccttggg agggtctcct ctgagtgatt 4801 gactacccgt cagcgggggt cttcacatgc agcatgtatc aaaattaatt tggttttttt 4861 tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa cgcgcgggga 4921 gaggcggttt gcgtattggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 4981 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 5041 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 5101 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 5161 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 5221 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggataccg 5281 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 5341 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 5401 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 5461 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 5521 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc 5581 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 5641 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 5701 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 5761 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 5821 ttaaattaaa aatgaagttt gcggccgcaa atcaatctaa agtatatatg agtaaacttg 5881 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 5941 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 6001 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 6061 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 6121 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 6181 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 6241 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 6301 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 6361 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 6421 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 6481 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt 6541 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 6601 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 6661 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 6721 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 6781 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 6841 aataggggtt ccgcgcacat ttccccgaaa agtgccac //