LOCUS Exported 8041 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGC180 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8041) AUTHORS Voutev R, Hubbard EJ TITLE A ""FLP-Out"" system for controlled gene expression in Caenorhabditis elegans. JOURNAL Genetics. 2008 Sep . 180(1):103-19. PUBMED 18723890 REFERENCE 2 (bases 1 to 8041) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8041 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 19..38 /label=T7 /note="T7 promoter, forward primer" promoter 19..37 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 60..184 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 209..239 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 293..952 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(359..378) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 1294..1599 /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 1478..1497 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(1640..1764) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1780..1796) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1781..1797) /label=pBluescriptKS /note="For pBluescript vector" CDS 1820..3091 /codon_start=1 /product="site-specific recombinase" /label=FLP /note="FLP is a site-specific recombinase from Saccharomyces cerevisiae. Recombination occurs at FRT sequences." /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR YPAWNGIISQEVLDYLSSYINRRI" CDS complement(3791..3802) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind 5957..5978 /label=F1ori-F /note="F1 origin, forward primer" promoter 6141..6245 /gene="bla" /label=AmpR promoter CDS 6246..7106 /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(6464..6483) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7277..7865 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7766..7785 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8019..8036 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 aacctggctt atcgaaatta atacgactca ctatagggag accggcagat ctgatatcaa 61 caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa 121 attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcat 181 attggcggcc gcattaggca ccccaggctt tacactttat gcttccggct cgtataatgt 241 gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta aaatggagaa 301 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga 361 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc 421 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct 481 tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg gtgagctggt 541 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc 601 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga 661 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt 721 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat 781 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt 841 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag 901 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aaagatctgg 961 atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt tttgcggtat 1021 aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc tatgaagcag 1081 cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata tatgatgtca 1141 atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac 1201 gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg 1261 gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta cacctataaa 1321 agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga cacgcccggg 1381 cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa 1441 ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac cgatatggcc 1501 agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg cgaaaatgac 1561 atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc ccttatacac 1621 agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt attatgtagt 1681 ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt acgtttctcg 1741 ttcagctttc ttgtacaaag tggttgatat caagcttatc gataccgtcg acctcgaggg 1801 ggggcccggt accaaaaaca tgccacaatt tgatatatta tgtaaaacac cacctaaggt 1861 gcttgttcgt cagtttgtgg aaaggtttga aagaccttca ggtgagaaaa tagcattatg 1921 tgctgctgaa ctaacctatt tatgttggat gattacacat aacggaacag caatcaagag 1981 agccacattc atgagctata atactatcat aagcaattcg ctgagtttcg atattgtcaa 2041 taaatcactc cagtttaaat acaagacgca aaaagcaaca attctggaag cctcattaaa 2101 gaaattgatt cctgcttggg aatttacaat tattccttac tatggacaaa aacatcaatc 2161 tgatatcact gatattgtaa gtagtttgca attacagttc gaatcatcgg aagaagcaga 2221 taagggaaat agccacagta aaaaaatgct taaagcactt ctaagtgagg gtgaaagcat 2281 ctgggagatc actgagaaaa tactaaattc gtttgagtat acttcgagat ttacaaaaac 2341 aaaaacttta taccaattcc tcttcctagc tactttcatc aattgtggaa gattcagcga 2401 tattaagaac gttgatccga aatcatttaa attagtccaa aataagtatc tgggagtaat 2461 aatccagtgt ttagtgacag agacaaagac aagcgttagt aggcacatat acttctttag 2521 cgcaaggggt aggatcgatc cacttgtata tttggatgaa tttttgagga attctgaacc 2581 agtcctaaaa cgagtaaata ggaccggcaa ttcttcaagc aataaacagg aataccaatt 2641 attaaaagat aacttagtca gatcgtacaa taaagctttg aagaaaaatg cgccttattc 2701 aatctttgct ataaaaaatg gcccaaaatc tcacattgga agacatttga tgacctcatt 2761 tctttcaatg aagggcctaa cggagttgac taatgttgtg ggaaattgga gcgataagcg 2821 tgcttctgcc gtggccagga caacgtatac tcatcagata acagcaatac ctgatcacta 2881 cttcgcacta gtttctcggt actatgcata tgatccaata tcaaaggaaa tgatagcatt 2941 gaaggatgag actaatccaa ttgaggagtg gcagcatata gaacagctaa agggtagtgc 3001 tgaaggaagc atacgatacc ccgcatggaa tgggataata tcacaggagg tactagacta 3061 cctttcatcc tacataaata gacgcatata agtacgcatt taagcataaa cacgcactat 3121 gccgttcttc tcatgtatat atatatacag gcaacacgca gatataggtg cgacgtgaac 3181 agtgagctgt atgtgcgggg atccactagt tctagccggc catacaagta atccggatga 3241 tcgacgccaa cgtcgttgaa ttttcaaatt ttaaatactg aatatttgtt ttttttccta 3301 ttatttattt attctctttg tgtttttttt cttgctttct aaaaaattaa ttcaatccaa 3361 atctaaacat gagcggtttt ttttctcttt ccgtctccca attcgtattc cgctcctctc 3421 atctgaacac aatgtgcaag tttatttatc ttctcgcttt catttcatta ggacgtgggg 3481 ggaattggtg gaagggggaa acacacaaaa ggatgatgga aatgaaataa ggacacacaa 3541 tatgcaacaa cattcaattc agaaatatgg aggaaggttt aaaagaaaac ataaaaatat 3601 atagaggagg aaggaaaact agtctagaat cctatgcttg cactttgagc caatttatcc 3661 aagtccttgt aaaaagtatt cgaaaattgt taacggataa aatgtttatt ataatatcaa 3721 aaacaatttg tcagttgacc actttttgat atatttgaca gaaaccggga tgaattggct 3781 caaagttagg gcgcccttct attacagggt ttctgataac aaaccggtta ttaactccca 3841 acaagggatg atttcaattc atcatgctca aattgaccca aattaagtta catgacaaat 3901 tcatcgccct tttcaactct tcttggctca tcatctgtta ttctgttcct attctgtctg 3961 caccctatac ccctttgcat actctctcgt cattcctttc ttggatagtg tcttcttctc 4021 cccagctctg ctacttctat gacttgcgcc cgctgctttt ccgctcgttc ctctcctctc 4081 tgacgtcttc ttctgcttct tttctccttc ctttttccct tctcaatctt cctctttttc 4141 ccatttctgt caatcattcg aagaagaaga agaagaagac cctcattcat ttcatttttt 4201 tttctgtcgg tgtgtgctgc tcggttaaga gtgagctctc tattccacgt cttcttcttt 4261 ttcttcttcg attcgaatca atcactccac caaaacgcat tcgtttttgg gattcacccc 4321 gcggttcgca acaggtttct ttttcaaatt attagcgtta taaatagaaa aatggtggag 4381 tttcaataaa aatgataatt tacaaagtgt attttgatta catgtactca aaaaggttga 4441 aatttttcat accagttttc cggaaatcca tctgatatca ttatcgtatt tttcttttta 4501 aaaatgtttt caaaaaaaaa caaaatatag ctggttattt ggcaccctct aattaccatt 4561 ttcctgtcac accacactct tttccttctt ccacttcttt ccgttttcag ccgcttccaa 4621 ccaaaccgat atgaaagccg agcaacaaca atcgattcca cccggctcgg cgaccttccc 4681 gtcgcaggtg agactcagaa aactagagaa accgctcaac taactcttga tatccgattt 4741 cattcttttc tcttttcttt ttgttgaact tttccatatt tccagatgcc acggccacca 4801 ccaagcaccg aacaaggaat cacaacggaa tcggagcttg cgaagaaagc tcaaatcact 4861 ccgaacgacg ttttagcact tccgggaatc actcaaggta tcttgctccc tttggatttc 4921 agtttaaaac ataaatttag gattcttatg ctccccatct gcgaacatct ataacatcga 4981 gttcaccaag ttccaaatcc gtgatctgga cactgagcaa gtgcttttcg agatcgccaa 5041 accggagaac gatcaggaga atgatgagtc gccacaggag tcggcaagat acgtgcgtta 5101 tagatttgct ccaaactttt tgaaactcaa aacggtcgga gcaactgtgg aattcaaagt 5161 aggagacatc ccaatccatc atttccgaat gatcgaacgt cacttcttca aagatcgcct 5221 tctgaagtgt tttgactttg aattcggatt ctgtattccg aattcacgaa acaactgtga 5281 acatatctat gagttccctc aactctctca acaactcagt gagtcattat tctaaaaagt 5341 acaattcaaa agactaatct ctttttcagt ggacgacatg atcaacaatc caaacgagac 5401 tcgttctgac agcttctatt tcgtcgataa caaactcgtc atgcacaaca aagccgacta 5461 ctcatatgat gcataaatat ttaatacaaa aatgttctgg ataattattc tgtcgaatag 5521 aaaaaaaact ccaaatgtga ttaaattcca ataattcctg tctagtttgt tccttcccct 5581 tccccttctc atgttcaatg cattcctaag cttttcagtt ccccccttgt ttctatattt 5641 tttcgctgtc ctgtcacact cgctaaaaac actaatcaca cggaaatctg ttttcaataa 5701 aaactccaac tttaactcat tttcaatttc aactgaaaga ttttttcatt agagaatgtc 5761 taggcatgct ctagagggcc caaaaggggc tttccccgtc aagctctaaa tcgggggctc 5821 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 5881 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 5941 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 6001 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag 6061 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg 6121 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 6181 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga 6241 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 6301 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 6361 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 6421 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 6481 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 6541 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 6601 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 6661 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 6721 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 6781 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc 6841 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc 6901 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 6961 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 7021 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 7081 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga 7141 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 7201 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 7261 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 7321 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 7381 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt 7441 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 7501 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 7561 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 7621 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 7681 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 7741 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 7801 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 7861 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 7921 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt 7981 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag 8041 c //