LOCUS Exported 4978 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGC244 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4978) 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 4978) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4978 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind complement(78..125) /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." protein_bind 199..323 /gene="mutant version of attR" /label=attR4 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS complement(364..669) /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 VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind complement(466..485) /label=ccdB-fwd /note="ccdB gene, forward primer" CDS complement(1011..1670) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 1585..1604 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(1724..1754) /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind complement(1780..1903) /gene="mutant version of attR" /label=attR3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2418..2440) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2540..2559 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(2637..2655) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2723..2827 /gene="bla" /label=AmpR promoter CDS 2828..3688 /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(3046..3065) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3859..4447 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4348..4367 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4601..4618 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4735..4756 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4771..4801 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4809..4825 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /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)." primer_bind 4814..4836 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4833..4849 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4833..4849 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 aagcttgcat gcctgcaggt cgactctaga ccgcggtttt ggcgcggatc cactagttct 61 agcttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga 121 acttcaagat ccccgggtat gtttcgaatg atactaacat aacatagaac attttcagga 181 gctcattacg ccaagctatc aactttgtat agaaaagttg aacgagaaac gtaaaatgat 241 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa 301 acacaacata tccagtcact atggtcgacc tgcagactgg ctgtgtataa gggagcctga 361 catttatatt ccccagaaca tcaggttaat ggcgtttttg atgtcatttt cgcggtggct 421 gagatcagcc acttcttccc cgataacgga gaccggcaca ctggccatat cggtggtcat 481 catgcgccag ctttcatccc cgatatgcac caccgggtaa agttcacggg ggactttatc 541 tgacagcaga cgtgcactgg ccagggggat caccatccgt cgcccgggcg tgtcaataat 601 atcactctgt acatccacaa acagacgata acggctctct cttttatagg tgtaaacctt 661 aaactgcatt tcaccagccc ctgttctcgt cggcaaaaga gccgttcatt tcaataaacc 721 gggcgacctc agccatccct tcctgatttt ccgctttcca gcgttcggca cgcagacgac 781 gggcttcatt ctgcatggtt gtgcttaccg aaccggagat attgacatca tatatgcctt 841 gagcaactga tagctgtcgc tgtcaactgt cactgtaata cgctgcttca tagcatacct 901 ctttttgaca tacttcgggt atacatatca gtatatattc ttataccgca aaaatcagcg 961 cgcaaatacg catactgtta tctggctttt agtaagccgg atcctctaga ttacgccccg 1021 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca 1081 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta 1141 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa 1201 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac 1261 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt 1321 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc 1381 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc 1441 attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc 1501 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga 1561 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga 1621 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc 1681 ttagctcctg aaaatctcga cggatcctaa ctcaaaatcc acacattata cgagccggaa 1741 gcataaagtg taaagcctgg gggtgcctaa tgcggccgcc atagtgactg gatatgttgt 1801 gttttacagt attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt 1861 atatcatttt acgtttctcg ttcaacttta ttatacatag ttgataattc actgggctag 1921 ccggccatac aagtaatccg gatgatcgac gccaacgtcg ttgaattttc aaattttaaa 1981 tactgaatat ttgttttttt tcctattatt tatttattct ctttgtgttt tttttcttgc 2041 tttctaaaaa attaattcaa tccaaatcta aacatgagcg gttttttttc tctttccgtc 2101 tcccaattcg tattccgctc ctctcatctg aacacaatgt gcaagtttat ttatcttctc 2161 gctttcattt cattaggacg tggggggaat tggtggaagg gggaaacaca caaaaggatg 2221 atggaaatga aataaggaca cacaatatgc aacaacattc aattcagaaa tatggaggaa 2281 ggtttaaaag aaaacataaa aatatataga ggaggaagga aaactagtaa aaaataagca 2341 aagaaattag gcgaacgatg agaattgtcc tcgcttgggc cctttcgtct cgcgcgtttc 2401 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg 2461 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt 2521 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg 2581 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcggcc ttaagggcct 2641 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg 2701 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 2761 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag 2821 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg 2881 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt 2941 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt 3001 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt 3061 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa 3121 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag 3181 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac 3241 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac 3301 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac 3361 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac 3421 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact 3481 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg 3541 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt 3601 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat 3661 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta 3721 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa 3781 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 3841 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 3901 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 3961 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc 4021 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 4081 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 4141 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 4201 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag 4261 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 4321 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 4381 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 4441 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 4501 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga 4561 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga 4621 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg 4681 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt 4741 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt 4801 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc 4861 caagctgtaa gtttaaacat gatcttacta actaactatt ctcatttaaa ttttcagagc 4921 ttaaaaatgg ctgaaatcac tcacaacgat ggatacgcta acaacttgga aatgaaat //