LOCUS Exported 5229 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION IL-6 promoter reporter construct. ACCESSION . VERSION . KEYWORDS pCR2.1-promoter-mIL-6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5229) AUTHORS Eitson JL, Medeiros JJ, Hoover AR, Srivastava S, Roybal KT, Ainsa JA, Hansen EJ, Gumbo T, van Oers NS TITLE Mycobacterial shuttle vectors designed for high-level protein expression in infected macrophages. JOURNAL Appl Environ Microbiol. 2012 Oct;78(19):6829-37. doi: 10.1128/AEM.01674-12. Epub 2012 Jul 20. PUBMED 22820329 REFERENCE 2 (bases 1 to 5229) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5229 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(208..227) /label=T7 /note="T7 promoter, forward primer" promoter complement(209..227) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(234..251) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(234..250) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(243..265) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 391..819 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(478..497) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 688..709 /label=F1ori-F /note="F1 origin, forward primer" CDS 1163..1957 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1217..1236) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1827..1846 /label=Neo-F /note="Neomycin resistance gene, forward primer" CDS 1975..2835 /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(2193..2212) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3006..3594 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3495..3514 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3748..3765 /label=L4440 /note="L4440 vector, forward primer" protein_bind 3882..3903 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3918..3948 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3956..3972 /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 3961..3983 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 3980..3996 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 3980..3996 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 ctctggcccc acccccaccc tccaacaaag atttttatca aatgtgggat tttcccatga 61 gtctcaaaat tagagagttg actcctaata aatatgagac tggggatgtc tgtagctcat 121 tctgctctgg accggtgcaa gggcgaattc tgcagatatc catcacactg gcggccgctc 181 gagcatgcat ctagagggcc caattcgccc tatagtgagt cgtattacaa ttcactggcc 241 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca 301 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc 361 caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg 421 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 481 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 541 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 601 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 661 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 721 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt 781 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaatt cagggcgcaa 841 gggctgctaa aggaagcgga acacgtagaa agccagtccg cagaaacggt gctgaccccg 901 gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca 961 ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag 1021 cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa 1081 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat ctgatcaaga 1141 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 1201 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 1261 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 1321 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac 1381 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 1441 attgggcgaa gtgccggggc aggatctcct gtcatcccac cttgctcctg ccgagaaagt 1501 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 1561 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 1621 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 1681 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 1741 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 1801 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 1861 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 1921 catcgccttc tatcgccttc ttgacgagtt cttctgaatt gaaaaaggaa gagtatgagt 1981 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 2041 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 2101 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 2161 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 2221 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 2281 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 2341 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 2401 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 2461 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 2521 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 2581 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 2641 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 2701 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 2761 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 2821 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 2881 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 2941 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 3001 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 3061 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 3121 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac 3181 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 3241 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 3301 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 3361 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 3421 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 3481 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 3541 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 3601 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 3661 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 3721 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc 3781 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac 3841 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact 3901 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg 3961 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttggtacc 4021 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgcccttg cattaatgat 4081 cctgagagtg tgttttgtaa atggttttgg attttatgta cagagcctac tttcagcctg 4141 gaatcattct gaatgctagc tagatatctg gagacaggtg gacagaaaac caggaactag 4201 tctgaaaaag aaactaacca aagggaagaa gtctgtttaa gtttgaccca gcctagaaga 4261 cttgagcatt ggaggggtta ttcagagtga gacgtaccac cttcagattc aaatcctgtc 4321 atccagtaga agggagcttc aaacacaagc tagctaagat acaatgaggt ccttcttcga 4381 tatctttatc ttccatatac catgaatcaa agaaacttca acaacatgag gactgcaaca 4441 gaccttcaag cctccttgca tgacctggaa atgttttggg gtgtcctggc agcagtggga 4501 tcagcactaa cagataaggg caactctcac agagactaaa ggtcttaact aagaagatag 4561 ccaagagacc actggggaga atgcagagaa taggcttgga cttgggagcc aagattgctt 4621 gacaacagac agaagatatt tctgtacttc acccacttta cccacctggc aactcctgga 4681 aacaactgca caaaatttgg aggtgaacaa accattagaa acaactggtc ctgacaagac 4741 acaggaaaaa caagcaatat gcaacattac tgtctgttgt ccaggttggg tgctgggggt 4801 gggagaggga gtgtgtgtct ttgtatgatc tgaaaaaact caggtcagaa catctgtaga 4861 tccttacaga catacaaaag aatcctagcc tcttattcat gtgtgtgtgt gtgtgtgtgt 4921 gtgtgtgtgt gtgtgtgtgt gtgtgtatgt gtgtgtcgtc tgtcatgcgc gcgtgcctgc 4981 gtttaaataa catcagcttt agcttctctt tctccttata aaacattgtg aatttcagtt 5041 ttctttccca tcaagacatg ctcaagtgct gagtcacttt taaagaaaaa aaagaagagt 5101 gctcatgctt cttagggcta gcctcaagga tgacttaagc acactttccc cttcctagtt 5161 gcgattcttt cgatgctaaa cgacgtcaca ttgtgcaatc ttaataaggt ttccaatcag 5221 ccccaccca //