LOCUS Exported 7698 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pQLinkGD SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7698) AUTHORS Scheich C, Kummel D, Soumailakakis D, Heinemann U, Bussow K TITLE Vectors for co-expression of an unrestricted number of proteins. JOURNAL Nucleic Acids Res. 2007 Feb 20. ():. PUBMED 17311810 REFERENCE 2 (bases 1 to 7698) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7698 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 36..80 /label=T5 promoter /note="bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator" protein_bind 56..72 /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)." protein_bind 88..104 /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)." RBS 123..134 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 141..794 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" primer_bind 752..774 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" protein_bind 816..940 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS complement(1369..1674) /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 complement(1471..1490) /label=ccdB-fwd /note="ccdB gene, forward primer" CDS complement(2019..2678) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" promoter complement(2679..2781) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" protein_bind complement(2959..3083) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" terminator 3105..3199 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS 3293..3952 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" terminator 4017..4103 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(4196..5278) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 5240..5259 /label=LacI-R /note="LacI, reverse primer" promoter complement(5279..5356) /gene="lacI (mutant)" /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." primer_bind complement(5440..5462) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 5548..5688 /label=bom /note="basis of mobility region from pBR322" primer_bind 5603..5622 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(5703..5720) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5874..6462) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5954..5973) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6633..7493) /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 7256..7275 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7494..7598) /gene="bla" /label=AmpR promoter primer_bind 7666..7684 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 ctcgagctta attaacaaca ccatttgtcg agaaatcata aaaaatttat ttgctttgtg 61 agcggataac aattataata gattcaattg tgagcggata acaatttcac acagaattca 121 ttaaagagga gaaattaact atgtccccta tactaggtta ttggaaaatt aagggccttg 181 tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg 241 agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca 301 atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt 361 atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa 421 tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag 481 actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg 541 aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca 601 tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc 661 caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga 721 aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg 781 accatcctcc aaaatcggat ctgggtgctg caggtacaag tttgtacaaa aaagctgaac 841 gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga 901 ctacataata ctgtaaaaca caacatatcc agtcactatg aatcaactac ttagatggta 961 ttagtgacct gtagtcgacc gacagccttc caaatgttct tcgggtgatg ctgccaactt 1021 agtcgaccga cagccttcca aatgttcttc tcaaacggaa tcgtcgtatc cagcctactc 1081 gctattgtcc tcaatgccgt attaaatcat aaaaagaaat aagaaaaaga ggtgcgagcc 1141 tcttttttgt gtgacaaaat aaaaacatct acctattcat atacgctagt gtcatagtcc 1201 tgaaaatcat ctgcatcaag aacaatttca caactcttat acttttctct tacaagtcgt 1261 tcggcttcat ctggattttc agcctctata cttactaaac gtgataaagt ttctgtaatt 1321 tctactgtat cgacctgcag actggctgtg tataagggag cctgacattt atattcccca 1381 gaacatcagg ttaatggcgt ttttgatgtc attttcgcgg tggctgagat cagccacttc 1441 ttccccgata acggagaccg gcacactggc catatcggtg gtcatcatgc gccagctttc 1501 atccccgata tgcaccaccg ggtaaagttc acgggagact ttatctgaca gcagacgtgc 1561 actggccagg gggatcacca tccgtcgccc gggcgtgtca ataatatcac tctgtacatc 1621 cacaaacaga cgataacggc tctctctttt ataggtgtaa accttaaact gcatttcacc 1681 agcccctgtt ctcgtcagca aaagagccgt tcatttcaat aaaccgggcg acctcagcca 1741 tcccttcctg attttccgct ttccagcgtt cggcacgcag acgacgggct tcattctgca 1801 tggttgtgct taccagaccg gagatattga catcatatat gccttgagca actgatagct 1861 gtcgctgtca actgtcactg taatacgctg cttcatagca tacctctttt tgacatactt 1921 cgggtataca tatcagtata tattcttata ccgcaaaaat cagcgcgcaa atacgcatac 1981 tgttatctgg cttttagtaa gccggatcca cgcggcgttt acgccccgcc ctgccactca 2041 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acagacggca 2101 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc 2161 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg 2221 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa 2281 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg 2341 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg 2401 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg 2461 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg 2521 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta 2581 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat 2641 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa 2701 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg 2761 gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg 2821 gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt 2881 tattcggcgc aaagtgcgtc gggtgatgct gccaacttag tcgactacag gtcactaata 2941 ccatctaagt agttgattca tagtgactgg atatgttgtg ttttacagta ttatgtagtc 3001 tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt 3061 tcagctttct tgtacaaagt ggtaagctta attagctgag cttggactcc tgttgataga 3121 tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc 3181 gttttttatt ggtgagaatc caagtaacaa caccatttaa atggagtggt tacaaatgga 3241 gtggttaatt aaggctagct tggcgagatt ttcaggagct aaggaagcta aaatggagaa 3301 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga 3361 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc 3421 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct 3481 tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt 3541 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc 3601 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga 3661 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt 3721 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat 3781 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt 3841 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag 3901 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta 3961 aggcagttat tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa 4021 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga 4081 acgctctcct gagtaggaca aatccgccct ctagattacg tgcagtcgat gataagctgt 4141 caaacatgag aattgtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact 4201 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 4261 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg 4321 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc 4381 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg 4441 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg 4501 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag 4561 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc 4621 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc 4681 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct 4741 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa 4801 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc 4861 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac 4921 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt 4981 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga 5041 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact 5101 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg 5161 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa 5221 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat 5281 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc 5341 accattcgat ggtgtcggaa tttcgggcag cgttgggtcc tggccacggg tgcgcatgat 5401 ctagagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc 5461 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc 5521 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg 5581 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccacatg 5641 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct 5701 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 5761 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 5821 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 5881 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 5941 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 6001 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 6061 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 6121 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 6181 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 6241 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 6301 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 6361 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 6421 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 6481 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 6541 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 6601 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 6661 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 6721 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 6781 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 6841 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 6901 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 6961 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 7021 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 7081 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 7141 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 7201 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 7261 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 7321 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 7381 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 7441 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 7501 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 7561 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 7621 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg 7681 aggccctttc gtcttcac //