LOCUS Exported 4833 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGC296 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4833) 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 4833) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4833 /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)." CDS 202..222 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 244..264 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 286..306 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 328..348 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(2273..2295) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2395..2414 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(2492..2510) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2578..2682 /gene="bla" /label=AmpR promoter CDS 2683..3543 /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(2901..2920) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3714..4302 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4203..4222 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4456..4473 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4590..4611 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4626..4656 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4664..4680 /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 4669..4691 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4688..4704 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4688..4704 /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 ccccggggta tgtttcgaat gatactaaca taacatagaa cattttcagg 181 agctcagaaa aaatgactgc tccaaagaag aagcgtaagg taccggcaga aaaaatgact 241 gctccaaaga agaagcgtaa ggtaccggca gaaaaaatga ctgctccaaa gaagaagcgt 301 aaggtaccgg cagaaaaaat gactgctcca aagaagaagc gtaaggtacc ggtagacgat 361 aaggatccga tggtggcctc ctccgaggac gtcatcaaag agttcatgcg cttcaaggtg 421 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc 481 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc 541 gcctgggaca tcctgtcccc ccagttccag tacggctcca aggcgtacgt gaagcacccc 601 gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg 661 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc 721 acgctgatct acaaggtgaa gttccgcggc accaacttcc cccccgacgg ccccgtaatg 781 cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg 841 ctgaagggcg agatccacca ggccctgaag ctgaaggacg gcggccacta cctggtggag 901 ttcaagacca tctacatggc caagaagccc gtgcagctgc ccggctacta ctacgtggac 961 accaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc 1021 tccgagggcc gccaccacct gttcctgggg catggcaccg gcagcaccgg cagcggcagc 1081 tccggcaccg cctcctccga ggacgtcatc aaagagttca tgcgcttcaa ggtgcgcatg 1141 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac 1201 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg 1261 gacatcctgt ccccccagtt ccagtacggc tccaaggcgt acgtgaagca ccccgccgac 1321 atccccgact acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac 1381 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcacgctg 1441 atctacaagg tgaagttccg cggcaccaac ttcccccccg acggccccgt aatgcagaag 1501 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag 1561 ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag 1621 accatctaca tggccaagaa gcccgtgcag ctgcccggct actactacgt ggacaccaag 1681 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag 1741 ggccgccacc acctgttcct gtaggaattc gctagccggc catacaagta atccggatga 1801 tcgacgccaa cgtcgttgaa ttttcaaatt ttaaatactg aatatttgtt ttttttccta 1861 ttatttattt attctctttg tgtttttttt cttgctttct aaaaaattaa ttcaatccaa 1921 atctaaacat gagcggtttt ttttctcttt ccgtctccca attcgtattc cgctcctctc 1981 atctgaacac aatgtgcaag tttatttatc ttctcgcttt catttcatta ggacgtgggg 2041 ggaattggtg gaagggggaa acacacaaaa ggatgatgga aatgaaataa ggacacacaa 2101 tatgcaacaa cattcaattc agaaatatgg aggaaggttt aaaagaaaac ataaaaatat 2161 atagaggagg aaggaaaact agtaaaaaat aagcaaagaa attaggcgaa cgatgagaat 2221 tgtcctcgct tgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac 2281 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag 2341 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat 2401 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 2461 ggagaaaata ccgcatcagg cggccttaag ggcctcgtga tacgcctatt tttataggtt 2521 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc 2581 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 2641 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc 2701 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 2761 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 2821 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg 2881 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa 2941 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc 3001 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc 3061 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta 3121 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 3181 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca 3241 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata 3301 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc 3361 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca 3421 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca 3481 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg 3541 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa 3601 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 3661 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 3721 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 3781 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 3841 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac 3901 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 3961 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 4021 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 4081 gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag 4141 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 4201 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 4261 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 4321 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 4381 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc 4441 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa 4501 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac 4561 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc 4621 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa 4681 tttcacacag gaaacagcta tgaccatgat tacgccaagc tgtaagttta aacatgatct 4741 tactaactaa ctattctcat ttaaattttc agagcttaaa aatggctgaa atcactcaca 4801 acgatggata cgctaacaac ttggaaatga aat //