LOCUS Exported 3905 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGC380 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3905) 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 3905) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3905 /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)." regulatory 139..148 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 145..822 /codon_start=1 /product="improved tetrameric variant of DsRed fluorescent protein" /label=DsRed2 /note="mammalian codon-optimized" /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH LFL" primer_bind complement(325..345) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 737..760 /label=DsRed1-C /note="DsRed1, forward primer" primer_bind complement(1345..1367) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 1467..1486 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(1564..1582) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1650..1754 /gene="bla" /label=AmpR promoter CDS 1755..2615 /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(1973..1992) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2786..3374 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3275..3294 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3528..3545 /label=L4440 /note="L4440 vector, forward primer" protein_bind 3662..3683 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3698..3728 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3736..3752 /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 3741..3763 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 3760..3776 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 3760..3776 /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 ccccggtcgc caccatggcc tcctccgaga acgtcatcac cgagttcatg 181 cgcttcaagg tgcgcatgga gggcaccgtg aacggccacg agttcgagat cgagggcgag 241 ggcgagggcc gcccctacga gggccacaac accgtgaagc tgaaggtgac caagggcggc 301 cccctgccct tcgcctggga catcctgtcc ccccagttcc agtacggctc caaggtgtac 361 gtgaagcacc ccgccgacat ccccgactac aagaagctgt ccttccccga gggcttcaag 421 tgggagcgcg tgatgaactt cgaggacggc ggcgtggcga ccgtgaccca ggactcctcc 481 ctgcaggacg gctgcttcat ctacaaggtg aagttcatcg gcgtgaactt cccctccgac 541 ggccccgtga tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc 601 cgcgacggcg tgctgaaggg cgagacccac aaggccctga agctgaagga cggcggccac 661 tacctggtgg agttcaagtc catctacatg gccaagaagc ccgtgcagct gcccggctac 721 tactacgtgg acgccaagct ggacatcacc tcccacaacg aggactacac catcgtggag 781 cagtacgagc gcaccgaggg ccgccaccac ctgttcctgt agcggccgcg actctagaat 841 tcgctagccg gccatacaag taatccggat gatcgacgcc aacgtcgttg aattttcaaa 901 ttttaaatac tgaatatttg ttttttttcc tattatttat ttattctctt tgtgtttttt 961 ttcttgcttt ctaaaaaatt aattcaatcc aaatctaaac atgagcggtt ttttttctct 1021 ttccgtctcc caattcgtat tccgctcctc tcatctgaac acaatgtgca agtttattta 1081 tcttctcgct ttcatttcat taggacgtgg ggggaattgg tggaaggggg aaacacacaa 1141 aaggatgatg gaaatgaaat aaggacacac aatatgcaac aacattcaat tcagaaatat 1201 ggaggaaggt ttaaaagaaa acataaaaat atatagagga ggaaggaaaa ctagtaaaaa 1261 ataagcaaag aaattaggcg aacgatgaga attgtcctcg cttgggccct ttcgtctcgc 1321 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc 1381 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg 1441 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca 1501 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta 1561 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga 1621 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 1681 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 1741 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 1801 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 1861 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 1921 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 1981 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 2041 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 2101 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 2161 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 2221 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 2281 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 2341 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 2401 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 2461 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 2521 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 2581 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 2641 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 2701 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 2761 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 2821 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 2881 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag 2941 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 3001 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 3061 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 3121 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt 3181 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 3241 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 3301 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 3361 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 3421 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 3481 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 3541 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc 3601 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa 3661 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc 3721 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg 3781 attacgccaa gctgtaagtt taaacatgat cttactaact aactattctc atttaaattt 3841 tcagagctta aaaatggctg aaatcactca caacgatgga tacgctaaca acttggaaat 3901 gaaat //