LOCUS Exported 7339 bp ds-DNA circular SYN 13-DEC-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS xCIAR_pcDNA5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7339) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 13, 2016 from SnapGene Viewer 3.0.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7339 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 235..614 /note="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /note="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" misc_feature 769..790 /label=CMV forward priming site /note="CMV forward priming site" misc_feature 804..810 /label=TATA box /note="TATA box" protein_bind 820..838 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" misc_feature 820..838 /label=Tet operator #1 /note="Tet operator #1" protein_bind 841..859 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tet operator" /note="bacterial operator O2 for the tetR and tetA genes" misc_feature 841..859 /label=Tet operator #2 /note="Tet operator #2" misc_feature 975 /label=Insertion site (flanking residues) /note="Insertion site (flanking residues)" misc_feature 976..978 /label=Start codon /note="Start codon" misc_feature join(979..1032,1036..1059,1111..2178,2182..3099) /label=xCIAR(aka xS.17.3) /note="xCIAR(aka xS.17.3)" misc_feature join(979..1032,1036..1059) /label=BAD /note="BAD" misc_feature 1021..1023 /label=L151 /note="L151" misc_feature 1033..1035 /label=S155A /note="S155A" misc_feature 1035 /label=SOS_17-6_T968L_F158G(1) /note="SOS_17-6_T968L_F158G(1)" misc_feature 1035 /label=BAD(1) /note="BAD(1)" misc_feature 1042..1044 /label=F158G /note="F158G" misc_feature 1060..1110 /label=N-term linker /note="N-term linker" misc_feature 2179..2181 /label=SOS_17-6_T968L_F158G /note="SOS_17-6_T968L_F158G" misc_feature 2179..2181 /label=F929A /note="F929A" misc_feature 2181 /label=F929 /note="F929" misc_feature 2296..2298 /label=T968L /note="T968L" misc_feature 2449..2457 /label=C-term linker /note="C-term linker" misc_feature 2458..3099 /label=BCL-XL_res_2-215 /note="BCL-XL_res_2-215" misc_feature 3100..3177 /label=HRAS_CaaX(c-term-25-residues) /note="HRAS_CaaX(c-term-25-residues)" misc_feature 3175..3177 /label=Stop /note="Stop" misc_feature 3178 /label=Insertion site (flanking residues)(1) /note="Insertion site (flanking residues)(1)" misc_feature 3291..3308 /label=BGH Reverse priming site /note="BGH Reverse priming site" polyA_signal 3297..3521 /note="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" protein_bind 3805..3852 /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FRT" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 3860..4882 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /note="HygR" /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 5012..5133 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" primer_bind complement(5182..5198) /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind 5206..5222 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(5230..5260) /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 5275..5296 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5584..6172) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6343..7203) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(7204..7308) /gene="bla" /note="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc 841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga 901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga 961 ctctagcgtt taaacatggc tcctcctaat ctgtgggcgg cacagcgtta tgggcgtgaa 1021 ctgcggcgta tggccgatga gggtgagggt tccttcaagg gtagcggaac tgggtctggg 1081 actgggagtg gcaccggttc agggacagga gatgtgtatc gctttgctga gcctgactcc 1141 gaagagaaca ttatcttcga agagaatatg cagcctaaag ctggtattcc gattatcaaa 1201 gccgggaccg tgattaagct gatcgagcgt ctgacatatc acatgtacgc cgaccccaac 1261 tttgttcgca ccttcctgac cacatatcgt tccttttgca agccacagga actgctgtct 1321 ctgattatcg agcggttcga aattccagaa ccagagccaa ccgaggcaga tcgtattgcg 1381 atcgaaaacg gcgaccagcc tctgagcgcg gagctgaaac gttttcggaa ggaatacatc 1441 cagccggtcc agctgcgggt tctgaatgtc tgtcgccatt gggtggagca ccatttttat 1501 gatttcgaac gggacgctta cctgctgcag cgcatggaag agttcattgg tacagtccgc 1561 gggaaagcca tgaaaaagtg ggtggaatcc attactaaga ttatccagcg caaaaagatc 1621 gcccgtgata acgggccagg acacaatatc acatttcaga gctccccgcc cactgtggag 1681 tggcacattt ctcgtcccgg tcatatcgaa accttcgatc tgctgacact gcatccaatt 1741 gagatcgctc gtcagctgac actgctggaa tctgacctgt atcgtgcagt gcagccaagc 1801 gagctggtgg gctccgtttg gaccaaagag gataaggaaa ttaacagtcc aaatctgctg 1861 aaaatgatcc gccacactac caacctgact ctgtggtttg aaaagtgcat tgttgaaacc 1921 gagaatctgg aagagcgtgt cgcggtggtt agccggatta tcgagatcct gcaggtgttt 1981 caggaactga acaatttcaa cggcgttctg gaggtcgtgt ccgcaatgaa cagcagccct 2041 gtgtatcgcc tggaccatac cttcgaacag attccgagcc gtcagaaaaa gatcctggaa 2101 gaggcgcacg agctgtccga agatcattat aaaaagtacc tggcaaaact gcgctctatt 2161 aacccacctt gcgtgccggc tttcggtatt tacctgacta acatcctgaa gaccgaagag 2221 ggaaatccgg aggttctgaa acgtcacggc aaggaactga tcaacttttc aaaacgccgt 2281 aaggtcgccg agattctcgg tgaaatccag cagtatcaga atcagcctta ctgtctgcgt 2341 gtggagagcg acattaaacg gtttttcgaa aacctgaatc cgatggggaa cagtatggaa 2401 aaagagttta ccgattacct gttcaataag tcactggaaa tcgagcctgg ctctggttca 2461 cagagcaacc gtgaactggt tgtcgatttt ctgtcctata aactgtctca gaagggctac 2521 tcctggtctc agttcagcga cgtcgaagag aatcggacag aggctccgga aggtactgaa 2581 tctgagatgg aaacccccag tgccattaac gggaatccat catggcacct ggcagatagc 2641 cccgctgtta atggagccac cggacattcg agctccctgg acgcccgcga ggttatccca 2701 atggccgcag tcaaacaggc gctgcgtgaa gcaggagatg agtttgaact gcgctatcgg 2761 cgcgcgttca gtgacctgac ctcacagctg cacattacac ccggcactgc ataccagagt 2821 tttgagcagg tggttaacga actgttccgc gacggagtga attggggccg tatcgttgcg 2881 tttttcagtt ttggcggtgc actgtgtgtc gagtcagtgg ataaagaaat gcaggtcctg 2941 gtgtctcgta ttgcagcttg gatggcaact tatctgaacg atcatctgga gccgtggatt 3001 caggaaaatg ggggatggga caccttcgtg gaactgtacg gcaacaatgc ggcggctgag 3061 agtcgtaagg ggcaggagcg gttcaatcgt tggtttctgc agcacaagct gcggaagctg 3121 aaccctcctg atgagagtgg ccccggctgc atgagctgca agtgtgtgct ctcctgatta 3181 agcttggtac cgagctcgga tccactagtc cagtgtggtg gaattctgca gatatccagc 3241 acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt 3301 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 3361 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 3421 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 3481 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 3541 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 3601 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 3661 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 3721 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 3781 ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag 3841 tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct 3901 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg 3961 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga 4021 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc 4081 ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc 4141 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt 4201 cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc 4261 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc 4321 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc 4381 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt 4441 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat 4501 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg 4561 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga 4621 gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta 4681 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc 4741 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc 4801 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac 4861 tcgtccgagg gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta 4921 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 4981 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 5041 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 5101 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 5161 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 5221 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 5281 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 5341 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 5401 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 5461 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 5521 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 5581 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 5641 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 5701 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 5761 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 5821 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 5881 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 5941 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6001 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 6061 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 6121 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 6181 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 6241 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 6301 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 6361 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 6421 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 6481 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 6541 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 6601 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 6661 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 6721 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 6781 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 6841 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 6901 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 6961 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7021 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 7081 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 7141 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 7201 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 7261 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 7321 aaaagtgcca cctgacgtc //