LOCUS Exported 5860 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial expression of Ubiquitin K6C for cross linking studies. ACCESSION . VERSION . KEYWORDS Ubiquitin K6C SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5860) AUTHORS Liu J, Chen Q, Rozovsky S TITLE Utilizing Selenocysteine for Expressed Protein Ligation and Bioconjugations. JOURNAL J Am Chem Soc. 2017 Feb 10. doi: 10.1021/jacs.6b10991. PUBMED 28186733 REFERENCE 2 (bases 1 to 5860) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5860 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 3..80 /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." CDS 81..1163 /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 complement(100..119) /label=LacI-R /note="LacI, reverse primer" promoter 1406..1434 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 1442..1458 /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 1447..1469 /label=M13/pUC Reverse /note="In lacZ gene" CDS 1537..1554 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind 2620..2643 /label=MBP-F /note="Maltose binding protein, forward primer" CDS 2659..2679 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" terminator 2946..3032 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 3124..3151 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 3179..3269 /gene="bla" /label=AmpR promoter CDS 3270..4130 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAWQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3488..3507) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4218..4806 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4707..4726 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4960..4977 /label=L4440 /note="L4440 vector, forward primer" primer_bind 5160..5182 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(5176..5367) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" ORIGIN 1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga 61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg 121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa 181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac 241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc 301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg 361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc 421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca 481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga 541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc 601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg 661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag 721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga 781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa 841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatttcggta gtgggatacg 901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc 961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga 1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata 1081 cacaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag 1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg 1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg 1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt 1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga 1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga 1501 gcaattgacc aacaaggacc atagattatg aaaatccatc accatcacca tcacgaagaa 1561 ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt 1621 aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa 1681 gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac 1741 gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg 1801 ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt 1861 gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac 1921 ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag 1981 agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac 2041 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat 2101 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg 2161 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg 2221 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta 2281 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca 2341 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg 2401 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg 2461 aagtcttacg aggaagagtt ggtgaaagat ccgcgtattg ccgccactat ggaaaacgcc 2521 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt 2581 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg 2641 cagactaatt cgagctctga aaacctgtat tttcagtccg ctcagatctt cgtctgcacg 2701 ttaaccggta aaaccataac tctggaagtt gaaccatccg ataccatcga aaacgttaag 2761 gctaaaattc aagacaagga aggaattcca cctgatcaac aaagattgat ctttgccggt 2821 aagcagctcg aggacggtag aacgctgtct gattacaaca ttcagaagga gtcgacctta 2881 catcttgtct taagactaag aggtggttaa ggatccgaat tccctgcagg taattaaata 2941 agcttcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt 3001 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga 3061 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta 3121 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt tcggtccgtt 3181 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 3241 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 3301 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 3361 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 3421 gcggtaagat ccttgagagt tttcgccccg aagaacgttt cccaatgatg agcactttta 3481 aagttctgct atgtggcgcg gtattatccc gtgttgacgc ctggcaagag caactcggtc 3541 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 3601 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 3661 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 3721 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 3781 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 3841 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 3901 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 3961 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 4021 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 4081 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 4141 aagtttactc atatatactt tagattgatt tccttaggac tgagcgtcaa ccccgtagaa 4201 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 4261 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 4321 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 4381 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 4441 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 4501 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 4561 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 4621 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 4681 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 4741 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 4801 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 4861 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 4921 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 4981 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc 5041 atataaggtg cactgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga 5101 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc 5161 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg 5221 gtaaagctca tcagcgtggt cgtgcagcga ttcacagatg tctgcctgtt catccgcgtc 5281 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt 5341 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat 5401 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga 5461 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga 5521 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc 5581 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga 5641 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca 5701 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt 5761 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat 5821 catgcgcacc cgtggccagg acccaacgct gcccgaaatt //