LOCUS Exported 6412 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses truncated human Ornithine Decarboxylase Antizyme fused to SPM in bacterial cells for calcium-induced covalent coupling to Ornithine Decarboxylase.. ACCESSION . VERSION . KEYWORDS pET28a-His6-OAZ-GSY-SPM-Ctag SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6412) AUTHORS Scheu AHA, Lim SYT, Metzner FJ, Mohammed S, Howarth M. TITLE NeissLock provides an inducible protein anhydride for covalent targeting of endogenous proteins. JOURNAL Nat Commun 12, 717 (2021). REFERENCE 2 (bases 1 to 6412) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6412 /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)" CDS 100..117 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 1277..1294 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1347..1365) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1361..1408 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 1445..1900 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1532..1551) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1742..1763 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(1992..2807) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 2715..2734 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2799..2818 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 2929..3517 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3418..3437 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3671..3688 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3703..3842 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3768..3787) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3928..3950 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3944..4135) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4944..6026) /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 5988..6007 /label=LacI-R /note="LacI, reverse primer" promoter complement(6027..6104) /gene="lacI" /label=lacI promoter primer_bind 6310..6329 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacttc 121 tactcggacg atcgtttgaa tgtgaccgaa gagttgacgt ccaacgacaa aacacgtatc 181 ttgaatgtcc agtcccgtct gactgacgcc aaacgtatta attggcgcac tgtacttagc 241 ggtggcagtc tttacatcga aatcccaggg ggcgccttgc ctgaaggctc taaagattcg 301 tttgccgtgt tgttggagtt tgccgaggag cagttacgcg cagatcatgt gttcatctgt 361 ttccacaaga accgcgaaga tcgtgcagcc cttctgcgta ctttctcctt ccttggcttc 421 gagatcgtgc gtccagggca tccgctggtg cccaagcgtc ctgacgcttg ttttatggca 481 tatactttcg aaggctcgta tgatccactg gcattagatc ttgatggcga cggtatcgaa 541 acggtggcaa cgaaggggtt tgccggctcc ttattcgatc acacaaataa cggaattcgc 601 acagctactg gatgggtaag tgccgacgac ggcctgttgg tgcgcgacct gaatggcaac 661 gggattatcg acaatggcgc ggaacttttt ggagataaca cgaaattggc agacgggagt 721 tttgctaagc acgggtatgc ggcgttggcg gagttggata gtaacggaga caatatcatc 781 aatgcggcag atgcggcatt ccagactttg cgcgtgtggc aagatctgaa tcaggacggt 841 atttcccaag ctaatgagct tcgtacattg gaggagcttg gtatccagtc gttagacctg 901 gcatacaaag atgtcaacaa gaatttaggt aacggtaata ctttagctca gcaaggtagc 961 tacaccaaaa ccgacggaac cactgcaaaa atgggcgact tgttgctggc agcagataat 1021 ctgcactcgc gtttcaagga caaagtagaa cttactgcgg aacaggcaaa ggccgcaaat 1081 ctggccggta ttggtcgttt acgcgactta cgcgaggccg cagctctttc tggagatctg 1141 gctaatatgt taaaagccta ttcagccgcg gagaccaaag aagcccaatt ggctctgttg 1201 gataatttga ttcacaaatg ggccgagacg gacgggagcg gtgagccaga ggcttaagct 1261 tgctaacgca ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg 1321 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc 1381 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa 1441 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 1501 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 1561 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 1621 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 1681 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 1741 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 1801 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 1861 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa 1921 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 1981 tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca 2041 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg 2101 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca 2161 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca 2221 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt 2281 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc 2341 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa 2401 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct 2461 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt 2521 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc 2581 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca 2641 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct 2701 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa 2761 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta 2821 ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt 2881 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 2941 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 3001 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 3061 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt 3121 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 3181 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 3241 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 3301 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 3361 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 3421 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 3481 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 3541 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 3601 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 3661 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct 3721 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct 3781 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg 3841 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat 3901 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt 3961 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt 4021 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg 4081 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg 4141 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga 4201 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta 4261 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc 4321 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga 4381 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga 4441 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc 4501 acgttcgctc gcgtatcggt gattcattct gctaaccggt aaggcaaccc cgccagccta 4561 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga 4621 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga 4681 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa 4741 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga 4801 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc 4861 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct 4921 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 4981 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg 5041 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc 5101 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg 5161 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc 5221 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc 5281 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg 5341 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt 5401 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt 5461 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc 5521 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca 5581 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca 5641 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc 5701 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt 5761 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg 5821 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg 5881 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg 5941 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg 6001 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc 6061 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc 6121 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac 6181 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg 6241 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc 6301 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat 6361 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa at //