LOCUS pAviTag_MBP_SAG1 4324 bp ds-DNA circular 29-AUG-2017 DEFINITION Vector for Expresso(R) cloning to add C-terminal biotinylation and 6xHis tags, with rhamnose-inducible expression. ACCESSION . VERSION . KEYWORDS pAviTag C-His Kan SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2359) AUTHORS Lucigen TITLE Direct Submission JOURNAL Exported Friday, Oct 09, 2015 from SnapGene 2.8.2 http://www.snapgene.com COMMENT COMMENT COMMENT ApEinfo:methylated:1 FEATURES Location/Qualifiers promoter 1..119 /note="rhaB promoter" /note="promoter of the E. coli rhaBAD operon, conferring tight induction with L-rhamnose and repression with D-glucose in the presence of RhaR and RhaS (Giacalone et al., 2006)" /locus_tag="rhaB promoter" /ApEinfo_label="rhaB promoter" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" CDS 2190..2234 /codon_start=1 /product="peptide tag that allows for enzymatic biotinylation" /note="AviTag(TM)" /translation="GLNDIFEAQKIEWHE" /locus_tag="AviTag" /ApEinfo_label="AviTag" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" CDS 2235..2252 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" /locus_tag="6xHis tag" /ApEinfo_label="6xHis tag" /ApEinfo_fwdcolor="#a1bdff" /ApEinfo_revcolor="pink" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature join(1368..1707,1709..2144) /locus_tag="SAG1 31-289" /ApEinfo_label="SAG1 31-289" /ApEinfo_fwdcolor="#40a3ff" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature 186..1367 /locus_tag="6His-MBP-N10-TEV" /ApEinfo_label="6His-MBP-N10-TEV" /ApEinfo_fwdcolor="#ffcf61" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" rep_origin 2480..3067 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /locus_tag="pBR322 ori" /ApEinfo_label="pBR322 ori" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" CDS complement(3130..3321) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /note="rop" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH A DELYRSCLARFGDDGENL" /locus_tag="Rop protein" /ApEinfo_label="Rop protein" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="#ffa47c" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" CDS complement(3322..4137) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /note="KanR" /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY G KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPG K TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVG A SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVG I ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" /locus_tag="KanR" /ApEinfo_label="KanR" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="#a8ff85" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" promoter complement(4138..4228) /note="cat promoter" /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" /locus_tag="cat promoter" /ApEinfo_label="cat promoter" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="#bfbfbf" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" ORIGIN 1 caccacaatt cagcaaattg tgaacatcat cacgttcatc tttccctggt tgccaatggc 61 ccattttcct gtcagtaacg agaaggtcgc gaattcaggc gctttttaga ctggtcgtag 121 ggagaccaca acggtttccc tctagaaata attttgttta actataagaa ggagatatac 181 atatgGGTTC TTCTATGAAA ATCGAAGAAG GTAAACTGGT AATCTGGATT AACGGCGATA 241 AAGGCTATAA CGGTCTCGCT GAAGTCGGTA AGAAATTCGA GAAAGATACC GGAATTAAAG 301 TCACCGTTGA GCATCCGGAT AAACTGGAAG AGAAATTCCC ACAGGTTGCG GCAACTGGCG 361 ATGGCCCTGA CATTATCTTC TGGGCACACG ACCGCTTTGG TGGCTACGCT CAATCTGGCC 421 TGTTGGCTGA AATCACCCCG GACAAAGCGT TCCAGGACAA GCTGTATCCG TTTACCTGGG 481 ATGCCGTACG TTACAACGGC AAGCTGATTG CTTACCCGAT CGCTGTTGAA GCGTTATCGC 541 TGATTTATAA CAAAGATCTG CTGCCGAACC CGCCAAAAAC CTGGGAAGAG ATCCCGGCGC 601 TGGATAAAGA ACTGAAAGCG AAAGGTAAGA GCGCGCTGAT GTTCAACCTG CAAGAACCGT 661 ACTTCACCTG GCCGCTGATT GCTGCTGACG GGGGTTATGC GTTCAAGTAT GAAAACGGCA 721 AGTACGACAT TAAAGACGTG GGCGTGGATA ACGCTGGCGC GAAAGCGGGT CTGACCTTCC 781 TGGTTGACCT GATTAAAAAC AAACACATGA ATGCAGACAC CGATTACTCC ATCGCAGAAG 841 CTGCCTTTAA TAAAGGCGAA ACAGCGATGA CCATCAACGG CCCGTGGGCA TGGTCCAACA 901 TCGACACCAG CAAAGTGAAT TATGGTGTAA CGGTACTGCC GACCTTCAAG GGTCAACCAT 961 CCAAACCGTT CGTTGGCGTG CTGAGCGCAG GTATTAACGC CGCCAGTCCG AACAAAGAGC 1021 TGGCAAAAGA GTTCCTCGAA AACTATCTGC TGACTGATGA AGGTCTGGAA GCGGTTAATA 1081 AAGACAAACC GCTGGGTGCC GTAGCGCTGA AGTCTTACGA GGAAGAGTTG GCGAAAGATC 1141 CACGTATTGC CGCCACTATG GAAAACGCCC AGAAAGGTGA AATCATGCCG AACATCCCGC 1201 AGATGTCCGC TTTCTGGTAT GCCGTGCGTA CTGCGGTGAT CAACGCCGCC AGCGGTCGTC 1261 AGACTGTCGA TGAAGCCCTG AAAGACGCGC AGACTAATTC GAGCTCGAAC AACAACAACA 1321 ATAACAATAA CAACAACCTC GGGATCGAGG AAAACCTGta cttccaaTCG GATCCCCCTC 1381 TTGTTGCCAA TCAAGTTGTC ACCTGCCCAG ATAAAAAATC GACAGCCGCG GTCATTCTCA 1441 CACCGACGGA GAACCACTTC ACTCTCAAGT GCCCTAAAAC AGCGCTCACA GAGCCTCCCA 1501 CTCTTGCGTA CTCACCCAAC AGGCAAATCT GCCCAGCGGG TACTACAAGT AGCTGTACAT 1561 CAAAGGCTGT AACATTGAGC TCCTTGATTC CTGAAGCAGA AGATAGCTGG TGGACGGGGG 1621 ATTCTGCTAG TCTCGACACG GCAGGCATCA AACTCACAGT TCCAATCGAG AAGTTCCCCG 1681 TGACAACGCA GACGTTTGTG GTCGGTTgCA TCAAGGGAGA CGACGCACAG AGTTGTATGG 1741 TCACGGTGAC AGTACAAGCC AGAGCCTCAT CGGTCGTCAA TAATGTCGCA AGGTGCTCCT 1801 ACGGTGCAGA CAGCACTCTT GGTCCTGTCA AGTTGTCTGC GGAAGGACCC ACTACAATGA 1861 CCCTCGTGTG CGGGAAAGAT GGAGTCAAAG TTCCTCAAGA CAACAATCAG TACTGTTCCG 1921 GGACGACGCT GACTGGTTGC AACGAGAAAT CGTTCAAAGA TATTTTGCCA AAATTAACTG 1981 AGAACCCGTG GCAGGGTAAC GCTTCGAGTG ATAAGGGTGC CACGCTAACG ATCAAGAAGG 2041 AAGCATTTCC AGCCGAGTCA AAAAGCGTCA TTATTGGATG CACAGGGGGA TCGCCTGAGA 2101 AGCATCACTG TACCGTGAAA CTGGAGTTTG CCGGGGCTGC AGGGGCCCta agcttatcca 2161 ccccgccgac cccgagcact cctcctaccg gtctgaacga catcttcgag gctcagaaaa 2221 tcgaatggca cgaacatcat caccaccatc actaatagag cggccgccac cgctgagcaa 2281 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 2341 ggaactatat ccgggtaacg aattcaagct tgatatcatt caggacgagc ctcagactcc 2401 agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc tttcttcggt 2461 agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 2521 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 2581 ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc 2641 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 2701 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 2761 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 2821 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 2881 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 2941 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 3001 gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct 3061 atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt gattacattt 3121 gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag 3181 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc 3241 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc 3301 ggttttttcc tgtttggtca tttagaaaaa ctcatcgagc atcaagtgaa actgcaattt 3361 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga 3421 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac 3481 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga 3541 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt 3601 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg caccaaccaa 3661 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc cgttaaaagg 3721 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat 3781 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc ctgggatcgc 3841 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg 3901 cataaattcc gtcagccagt ttagcctgac catctcatct gtaacatcat tggcaacgct 3961 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat 4021 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc 4081 catgttggaa tttaatcgcg gcctcgagca agacgtttcc cgttgaatat ggctcatagc 4141 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg 4201 aaagttggaa cctcttacgt gccgatcaag tcaaaagcct ccggtcggag gcttttgact 4261 ttctgctatg gaggtcaggt atgatttaaa tggtcagtat tgagcgatat ctagagaatt 4321 cgtc //