LOCUS Exported 6619 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Plasmid encoding the bacterial expression of the chimeric HaloTag7-NanoLucCP fusion dubbed H-Luc. ACCESSION . VERSION . KEYWORDS pET51b(+)_H-Luc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6619) AUTHORS Hiblot J, Yu Q, Sabbadini MDB, Reymond L, Xue L, Schena A, Sallin O, Hill N, Griss R, Johnsson K TITLE Luciferases with Tunable Emission Wavelengths. JOURNAL Angew Chem Int Ed Engl. 2017 Nov 13;56(46):14556-14560. doi: 10.1002/anie.201708277. Epub 2017 Oct 9. PUBMED 28941028 REFERENCE 2 (bases 1 to 6619) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6619 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 12..467 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(99..118) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 309..330 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(582..1442) /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 1205..1224 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1443..1535) /gene="bla" /label=AmpR promoter rep_origin 1616..2204 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2105..2124 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2358..2375 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2390..2529 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(2455..2474) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2615..2637 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(2631..2822) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3394..4476) /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 4438..4457 /label=LacI-R /note="LacI, reverse primer" promoter complement(4477..4554) /gene="lacI" /label=lacI promoter primer_bind 4760..4779 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 4867..4886 /label=T7 /note="T7 promoter, forward primer" promoter 4867..4885 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 4886..4910 /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 4925..4947 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 4963..4986 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 4993..5007 /codon_start=1 /product="enterokinase recognition and cleavage site" /label=enterokinase site /translation="DDDDK" CDS 6481..6510 /codon_start=1 /product="10xHis affinity tag" /label=10xHis /translation="HHHHHHHHHH" primer_bind complement(6533..6551) /label=T7 Term /note="T7 terminator, reverse primer" terminator 6547..6594 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" ORIGIN 1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat 481 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 1021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 1081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 1141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 1201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 1261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 1321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 1381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 1441 atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg 1501 atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac 1561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 1621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 1681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 1741 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 1801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 1861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 1921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 1981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 2041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 2101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 2161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 2221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 2281 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 2341 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt 2401 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat 2461 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc 2521 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc 2581 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt 2641 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga 2701 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc 2761 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc 2821 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga 2881 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt 2941 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa 3001 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg 3061 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa 3121 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag 3181 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc 3241 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc tagtcatgcc ccgcgcccac 3301 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa 3361 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 3421 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 3481 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac 3541 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa 3601 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta 3661 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc 3721 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag 3781 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat 3841 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga 3901 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg 3961 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg 4021 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc 4081 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt 4141 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct 4201 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag 4261 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc 4321 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt 4381 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc 4441 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc 4501 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat 4561 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc 4621 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc 4681 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc 4741 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg 4801 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg 4861 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa 4921 taattttgtt taactttaag aaggagatat accatggcaa gctggagcca cccgcagttc 4981 gaaaagggtg cagatgacga cgacaaggta ccgcatggat ccgaaatcgg tactggcttt 5041 ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt 5101 ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg 5161 cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga cctgatcggt 5221 atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt ccgcttcatg 5281 gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca cgactggggc 5341 tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg tattgcattt 5401 atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc ccgcgagacc 5461 ttccaggcct tccgcaccag cggcgaccaa atgggccaga tcgaaaaaat ttttaaggtg 5521 gtgtaccctg tggatgatca tcactttaag gtgatcctgc actatggcac actggtaatc 5581 gacggggtta cgccgaacat gatcgactat ttcggacggc cgtatgaagg catcgccgtg 5641 ttcgacggca aaaagatcac tgtaacaggg accctgtgga acggcaacaa aattatcgac 5701 gagcgcctga tcaaccccga cggctccctg ctgttccgag taaccatcaa cggagtgacc 5761 ggctggcggc tgtgcgaacg cattctggcg ggtggcaccg gtggctctgg cggtaccggt 5821 ggctctatgg tcttcacact cgaagatttc gttggggact ggcgacagac agccggctac 5881 aacctggacc aagtccttga acagggaggt gtgtccagtt tgtttcagaa tctcggggtg 5941 tccgtaactc cgatccaaag gattgtcctg agcggtgaaa atgggctgaa gatcgacatc 6001 catgtcatca tcccgtatga aggtctggac gtcggccgca agctgatcat cgatcagaac 6061 gtttttatcg agggtacgct gccgatgggt gtcgtccgcc cgctgactga agtcgagatg 6121 gaccattacc gcgagccgtt cctgaatcct gttgaccgcg agccactgtg gcgcttccca 6181 aacgagctgc caatcgccgg tgagccagcg aacatcgtcg cgctggtcga agaatacatg 6241 gactggctgc accagtcccc tgtcccgaag ctgctgttct ggggcacccc aggcgttctg 6301 atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac 6361 atcggcccgg gtctgaatct gctgcaagaa gacaacccgg acctgatcgg cagcgagatc 6421 gcgcgctggc tgtctactct ggagatttcc ggtgctccgg gcttctcctc aatttccgct 6481 catcaccacc atcatcacca tcaccaccac taattaacct aggctgctgc caccgctgag 6541 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa 6601 ggaggaacta tatccggat //