LOCUS Exported 2939 bp ds-DNA circular SYN 10-JUL-2017 DEFINITION High-copy number vector with a kanamycin resistance marker for efficient blunt cloning of unstable sequences. ACCESSION AF532107 VERSION . KEYWORDS pSMART HCKan SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2939) AUTHORS Lucigen TITLE Direct Submission JOURNAL Exported Friday, Jul 21, 2017 from SnapGene 3.3.4 http://www.snapgene.com COMMENT Linearize at HincII to make blunt cloning vector. FEATURES Location/Qualifiers source 1..2939 /organism="synthetic DNA construct" /lab_host="Escherichia coli" /mol_type="other DNA" misc_feature 2..7 /label=BsmBI /note="color: #a6acb3; direction: RIGHT" misc_feature 9..12 /label=O /note="color: #ffff00" misc_feature complement(14..19) /label=BsaI /note="color: #a6acb3; direction: LEFT" misc_feature 20..41 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" /note="color: #99ccff" protein_bind 142..163 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /note="color: #31849b" promoter 178..208 /label=lac promoter /note="promoter for the E. coli lac operon" /note="This forward directional feature has 3 segments: 1: 178 .. 183 / #ffffff / -35 2: 184 .. 201 / #ffffff 3: 202 .. 208 / #ffffff / -10" protein_bind 216..232 /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)." /note="color: #31849b" CDS 268..945 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /note="color: #ff0000" /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" terminator 990..1061 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" /note="color: #ffffff" terminator 1077..1104 /label=T7Te terminator /note="phage T7 early transcription terminator" /note="color: #ffffff" misc_feature 1111..1131 /label=BioBrick suffix /note="universal suffix for all parts" /note="color: #a6ccff" misc_feature 1132..1137 /label=BsaI /note="color: #a6acb3; direction: RIGHT" misc_feature 1139..1142 /label=P /note="color: #ffff00" misc_feature complement(1144..1149) /label=BsmBI /note="color: #a6acb3; direction: LEFT" terminator 1216..1247 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" /note="color: #ff0000" promoter 1248..1350 /label=cat promoter /note="promoter of the E. coli cat gene" /note="color: #ffffff; direction: RIGHT" CDS 1351..2166 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /note="color: #ccffcc" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" terminator 2190..2217 /label=T7Te terminator /note="phage T7 early transcription terminator" /note="color: #ff0000" rep_origin complement(2229..2816) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" terminator 2838..2867 /label=T3Te terminator /note="phage T3 early transcription terminator" /note="color: #ff0000" ORIGIN 1 acgtctcggc aaagagaccg aattcgcggc cgcttctaga gcaatacgca aaccgcctct 61 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc 121 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt 181 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac 241 atactagaga aagaggagaa atactagatg gcttcctccg aagacgttat caaagagttc 301 atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt 361 gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt 421 ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct 481 tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc 541 aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac ccaggactcc 601 tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc 661 gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac 721 ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt 781 cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt 841 gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta caccatcgtt 901 gaacagtacg aacgtgctga aggtcgtcac tccaccggtg cttaataacg ctgatagtgc 961 tagtgtagat cgctactaga gccaggcatc aaataaaacg aaaggctcag tcgaaagact 1021 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct 1081 caccttcggg tgggcctttc tgcgtttata tactagtagc ggccgctgca gggtctctcc 1141 ctcgagacga agacgaattc tctagatatc gctcaatact gaccatttaa atcatacctg 1201 acctccatag cagaaagtca aaagcctccg accggaggct tttgacttga tcggcacgta 1261 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta 1321 tcgagatttt caggagctaa ggaagctaaa atgagccata ttcaacggga aacgtcttgc 1381 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc 1441 gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca 1501 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc 1561 aggctaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact 1621 cctgatgatg catggttact caccactgcg atcccaggga aaacagcatt ccaggtatta 1681 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg 1741 ttgcattcga ttcctgtttg taattgtcct tttaacggcg atcgcgtatt tcgtctcgct 1801 caggcgcaat cacgaatgaa taacggtttg gttggtgcga gtgattttga tgacgagcgt 1861 aatggctggc ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg 1921 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa 1981 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc 2041 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa 2101 tatggtattg ataatcctga tatgaataaa ttgcagtttc acttgatgct cgatgagttt 2161 ttctaatgag ggcccaaatg taatcacctg gctcaccttc gggtgggcct ttctgcgttg 2221 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg atgctcaagt 2281 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 2341 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 2401 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 2461 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 2521 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 2581 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 2641 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag 2701 ccagttacct cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 2761 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 2821 atcctttgat tttctaccga agaaaggccc acccgtgaag gtgagccagt gagttgattg 2881 cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgtt //