LOCUS Exported 5807 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses mGinger1 tagged N-terminally with an MHHHHHHGSG tag. ACCESSION . VERSION . KEYWORDS pET-mGinger1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5807) AUTHORS Wannier TM, Gillespie SK, Hutchins N, McIsaac RS, Wu SY, Shen Y, Campbell RE, Brown KS, Mayo SL TITLE Monomerization of far-red fluorescent proteins. JOURNAL Proc Natl Acad Sci U S A. 2018 Nov 27;115(48):E11294-E11301. doi: 10.1073/pnas.1807449115. Epub 2018 Nov 13. PUBMED 30425172 REFERENCE 2 (bases 1 to 5807) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5807 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 812..831 /label=LacI-R /note="LacI, reverse primer" promoter complement(851..928) /gene="lacI" /label=lacI promoter primer_bind 1134..1153 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 1241..1260 /label=T7 /note="T7 promoter, forward primer" promoter 1241..1259 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1260..1284 /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 1299..1321 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 1331..1348 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 2045..2068 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" primer_bind complement(2095..2113) /label=T7 Term /note="T7 terminator, reverse primer" terminator 2109..2156 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 2193..2648 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2280..2299) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2490..2511 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(2763..3623) /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 3386..3405 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3624..3716) /gene="bla" /label=AmpR promoter rep_origin 3797..4385 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4286..4305 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4539..4556 /label=L4440 /note="L4440 vector, forward primer" misc_feature 4571..4710 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(4636..4655) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4796..4818 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(4812..5003) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(5575..850) /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" ORIGIN 1 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa 61 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa 121 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg 181 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat 241 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca 301 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt 361 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg 421 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa 481 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga 541 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt 601 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa 661 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct 721 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca 781 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta 841 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc 901 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc 961 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat 1021 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc 1081 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg 1141 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt 1201 ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact cactataggg 1261 gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag 1321 atataccatg caccaccatc atcaccatgg cagcggcctg ctgaaagaaa atatgcatat 1381 caagatgtat atggaaggca ccgtgaatgg tcactacttt aagtgtaagg gtgaaggcga 1441 tggtaatcca ttcgctggca cacagtccat gcgtgttcat gttaccgagg gcgctccgct 1501 ccctttcgcc tttgacatcc tcgccccgtg ctgtgaatat ggctctaaga cctttgttcg 1561 ctatccagcg gatatcccgg actttttcaa acagtccttc ccggagggct tcacttggga 1621 acgtacgact acctacgagg acggtggtat cctgaccgcg caccaggaca ccagcctgga 1681 gggtaactgc ctgatttaca aggttaaggt tcatggcacg aatttcccgg ctgacggtcc 1741 ggttatgaaa aaggaaactt gtggttggga accgtccacc gagagcgttt acccggaaaa 1801 cggtgttttg cgcggtcgta atcacatggc gctcaaggtt ggtgatagcc acctccactg 1861 ccaccactcc acgacctacc gctctaaaaa ggcggaacgc gccctgatta tgccgccgcg 1921 ccacagcact gattactgtc tgcaaataac gagtcataag gatgatgagt acttcgagct 1981 gtacgaaacg tctgtggcgc gttactctga ctaatgagct ttcttgtaca aagtggtttc 2041 cgcttggagc cacccgcagt tcgaaaagta agtgattaac ctaggctgct gccaccgctg 2101 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga 2161 aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc 2221 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 2281 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 2341 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 2401 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 2461 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 2521 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg 2581 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt 2641 tacaatttct ggcggcacga tggcatgaga ttatcaaaaa ggatcttcac ctagatcctt 2701 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 2761 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 2821 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 2881 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 2941 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 3001 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 3061 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 3121 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 3181 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 3241 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 3301 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 3361 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 3421 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 3481 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 3541 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 3601 acacggaaat gttgaatact catactcttc ctttttcaat catgattgaa gcatttatca 3661 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 3721 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 3781 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 3841 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 3901 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt 3961 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 4021 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 4081 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 4141 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 4201 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 4261 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 4321 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 4381 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 4441 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt 4501 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag 4561 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca 4621 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg 4681 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg 4741 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg 4801 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa 4861 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc 4921 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg 4981 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg 5041 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat 5101 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag 5161 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag 5221 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc 5281 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc 5341 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc 5401 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg 5461 ctagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc 5521 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg 5581 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 5641 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg 5701 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct 5761 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcg //