LOCUS Exported 4392 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pSKDuet16 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4392) AUTHORS Ellila S, Jurvansuu JM, Iwai H TITLE Evaluation and comparison of protein splicing by exogenous inteins with foreign exteins in Escherichia coli. JOURNAL FEBS Lett. 2011 Oct 10. PUBMED 22001202 REFERENCE 2 (bases 1 to 4392) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4392 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 61..80 /label=T7 /note="T7 promoter, forward primer" promoter 61..79 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 80..104 /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)." CDS 160..177 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 187..204 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 1006..1050 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" primer_bind complement(1088..1106) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1102..1149 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(1379..2194) /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 2102..2121 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" promoter complement(2195..2286) /gene="bla" /label=AmpR promoter rep_origin complement(2302..3051) /direction=LEFT /label=RSF ori /note="Plasmids containing the RSF 1030 origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins." primer_bind complement(2312..2329) /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(2552..2571) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3247..4329) /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 4291..4310 /label=LacI-R /note="LacI, reverse primer" promoter complement(4330..15) /gene="lacI" /label=lacI promoter ORIGIN 1 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaaat 61 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt 121 tgtttaactt taataaggag atataccatg ggcagcagcc atcatcatca tcatcacagc 181 agcggcctgg tgccgcgcgg cagcctagga cagtacaaac ttatcctgaa cggtaaaacc 241 ctgaaaggtg aaaccaccac cgaagctgtt gacgctgcta ccgcggaaaa agttttcaaa 301 cagtacgcta acgacaacgg tgttgacggt gaatggacct acgacgacgc taccaaaacc 361 tacaccgtaa cggaaggatc ctgtttaagc tatgaaacgg aaatattgac agtagaatat 421 ggattattac cgattggtaa aattgtagaa aagcgcatcg aatgtactgt ttatagcgtt 481 gataataatg gaaatattta tacacaacct gtagcacaat ggcacgatcg cggagaacaa 541 gaggtgtttg agtattgttt ggaagatggt tcattgattc gggcaacaaa agaccataag 601 tttatgactg ttgatggtca aatgttgcca attgatgaaa tatttgaacg tgaattggat 661 ttgatgcggg ttgataattt gccgaatatc aaaatagcca cacgtaaata tttaggcaaa 721 caaaatgtct atgacattgg agttgagcgc gaccataatt ttgcactcaa aaatggcttc 781 atagcttcta attgtttcaa tggtacccag tacaaactta tcctgaacgg taaaaccctg 841 aaaggtgaaa ccaccaccga agctgttgac gctgctaccg cggaaaaagt tttcaaacag 901 tacgctaacg acaacggtgt tgacggtgaa tggacctacg acgacgctac caaaacctac 961 accgtaacgg aataagcttg gctgttttgg cggctcgagt ctggtaaaga aaccgctgct 1021 gcgaaatttg aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct 1081 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg 1141 ggttttttgc tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc 1201 aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg 1261 acgacaagct gacgaccggg tctccgcaag tggcactttt cggggaaatg tgcgcggaac 1321 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt 1381 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac 1441 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata 1501 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta 1561 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg 1621 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc 1681 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg 1741 cctgagcgag acgaaatacg cggtcgctgt taaaaggaca attacaaaca ggaatcgaat 1801 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt 1861 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat 1921 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta 1981 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca 2041 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat 2101 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc 2161 tagagcaaga cgtttcccgt tgaatatggc tcatactctt cctttttcaa tattattgaa 2221 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 2281 aacaaatagg catgcagcgc tcttccgctt cctcgctcac tgactcgcta cgctcggtcg 2341 ttcgactgcg gcgagcggtg tcagctcact caaaagcggt aatacggtta tccacagaat 2401 caggggataa agccggaaag aacatgtgag caaaaagcaa agcaccggaa gaagccaacg 2461 ccgcaggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 2521 agccagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 2581 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 2641 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgtt ggtatctcag ttcggtgtag 2701 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 2761 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 2821 gcagccattg gtaactgatt tagaggactt tgtcttgaag ttatgcacct gttaaggcta 2881 aactgaaaga acagattttg gtgagtgcgg tcctccaacc cacttacctt ggttcaaaga 2941 gttggtagct cagcgaacct tgagaaaacc accgttggta gcggtggttt ttctttattt 3001 atgagatgat gaatcaatcg gtctatcaag tcaacgaaca gctattccgt tactctagat 3061 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt 3121 gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct 3181 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg 3241 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 3301 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac 3361 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa 3421 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg 3481 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac 3541 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac 3601 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga 3661 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata 3721 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag 3781 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc 3841 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg 3901 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat 3961 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac 4021 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt 4081 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat 4141 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc 4201 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac 4261 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac 4321 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg 4381 aaaggttttg cg //