LOCUS Exported 5994 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of WT MIF in E. coli for protein production. ACCESSION . VERSION . KEYWORDS pET11b-MIF SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5994) TITLE Lolis Lab Plasmids REFERENCE 2 (bases 1 to 5994) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5994 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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 59..81 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" primer_bind complement(495..513) /label=T7 Term /note="T7 terminator, reverse primer" terminator 509..556 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(731..759) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" primer_bind complement(786..804) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 872..976 /gene="bla" /label=AmpR promoter CDS 977..1837 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1195..1214) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2008..2596 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2497..2516 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2750..2767 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2782..2921 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(2847..2866) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3007..3029 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3023..3214) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4526..5608) /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 5570..5589 /label=LacI-R /note="LacI, reverse primer" promoter complement(5609..5686) /gene="lacI" /label=lacI promoter primer_bind 5892..5911 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag atatacatat gccgatgttc atcgtaaaca ccaacgtgcc 121 ccgcgcctcc gtgccggacg ggttcctctc cgagctcacc cagcagctgg cgcaggccac 181 cggcaagccc ccccagtaca tcgcggtgca cgtggtcccg gaccagctca tggccttcgg 241 cggctccagc gagccgtgcg cgctctgcag cctgcacagc atcggcaaga tcggcggcgc 301 gcagaaccgc tcctacagca agctgctgtg cggcctgctg gccgagcgcc tgcgcatcag 361 cccggacagg gtctacatca actattacga catgaacgcg gccaatgtgg gctggaacaa 421 ctccaccttc gcctaagagc cgcaggatcc ggctgctaac aaagcccgaa aggaagctga 481 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt 541 cttgaggggt tttttgctga aaggaggaac tatatccgga tatcccgcaa gaggcccggc 601 agtaccggca taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg 661 atgagcgcat tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat 721 aaactaccgc attaaagctt atcgatgata agctgtcaaa catgagaatt cttgaagacg 781 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta 841 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 901 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 961 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 1021 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 1081 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 1141 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 1201 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 1261 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 1321 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 1381 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 1441 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 1501 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga 1561 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 1621 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 1681 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 1741 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 1801 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 1861 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 1921 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 1981 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 2041 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 2101 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct 2161 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 2221 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 2281 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 2341 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 2401 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 2461 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 2521 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 2581 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 2641 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 2701 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 2761 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat 2821 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag 2881 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac 2941 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga 3001 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc 3061 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat 3121 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg 3181 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc 3241 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg 3301 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc 3361 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag 3421 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg 3481 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg 3541 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg 3601 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga 3661 gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc 3721 tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag 3781 ttctccgcaa gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc 3841 gccggcttcc attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag 3901 gcagacaagg tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga 3961 ggcggcataa atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc 4021 cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc 4081 ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat 4141 ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc 4201 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc 4261 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg 4321 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat 4381 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga 4441 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag 4501 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 4561 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca 4621 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc 4681 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt 4741 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta 4801 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg 4861 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca 4921 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa 4981 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac 5041 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc 5101 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga 5161 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt 5221 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg 5281 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca 5341 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac 5401 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt 5461 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa 5521 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg 5581 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc 5641 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc 5701 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc 5761 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg 5821 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga 5881 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg 5941 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaat //