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" terminator 73..102 /label=T3Te terminator /note="phage T3 early transcription terminator" /note="color: #ff0000" rep_origin 124..711 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" terminator 723..750 /label=T7Te terminator /note="phage T7 early transcription terminator" /note="color: #ff0000" CDS complement(774..1589) /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" promoter complement(1590..1692) /label=cat promoter /note="promoter of the E. coli cat gene" /note="color: #ffffff; direction: LEFT" terminator 1693..1724 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" /note="color: #ff0000" misc_feature 1790..1795 /label=BsmBI /note="color: #a6acb3; direction: RIGHT" misc_feature 1797..1800 /label=X /note="color: #ffff00" misc_feature complement(1802..1807) /label=BsaI /note="color: #a6acb3; direction: LEFT" misc_feature 1808..1829 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" /note="color: #99ccff" protein_bind 1930..1951 /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 1966..1996 /label=lac promoter /note="promoter for the E. coli lac operon" /note="This forward directional feature has 3 segments: 1: 1966 .. 1971 / #ffffff / -35 2: 1972 .. 1989 / #ffffff 3: 1990 .. 1996 / #ffffff / -10" protein_bind 2004..2020 /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 2056..2733 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /note="color: #ff0000" /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" terminator 2778..2849 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" /note="color: #ffffff" terminator 2865..2892 /label=T7Te terminator /note="phage T7 early transcription terminator" /note="color: #ffffff" misc_feature 2899..2919 /label=BioBrick suffix /note="universal suffix for all parts" /note="color: #a6ccff" misc_feature 2920..2925 /label=BsaI /note="color: #a6acb3; direction: RIGHT" misc_feature 2927..2930 /label=Y /note="color: #ffff00" misc_feature complement(2932..2937) /label=BsmBI /note="color: #a6acb3; direction: LEFT" ORIGIN 1 aacgaattca agcttgatat cattcaggac gagcctcaga ctccagcgta actggactgc 61 aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc 121 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 181 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc 241 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 301 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 361 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 421 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 481 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 541 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 601 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 661 cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 721 aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc tcattagaaa 781 aactcatcga gcatcaagtg aaactgcaat ttattcatat caggattatc aataccatat 841 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg 901 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat 961 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc 1021 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta 1081 cgctcgtcat caaaatcact cgcaccaacc aaaccgttat tcattcgtga ttgcgcctga 1141 gcgagacgaa atacgcgatc gccgttaaaa ggacaattac aaacaggaat cgaatgcaac 1201 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct 1261 aatacctgga atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga 1321 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagcctg 1381 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct 1441 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg 1501 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag 1561 caagacgttt cccgttgaat atggctcatt ttagcttcct tagctcctga aaatctcgat 1621 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt 1681 acgtgccgat caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc 1741 aggtatgatt taaatggtca gtattgagcg atatctagag aattcgtcac gtctcgcacg 1801 agagaccgaa ttcgcggccg cttctagagc aatacgcaaa ccgcctctcc ccgcgcgttg 1861 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg 1921 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct 1981 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacat actagagaaa 2041 gaggagaaat actagatggc ttcctccgaa gacgttatca aagagttcat gcgtttcaaa 2101 gttcgtatgg aaggttccgt taacggtcac gagttcgaaa tcgaaggtga aggtgaaggt 2161 cgtccgtacg aaggtaccca gaccgctaaa ctgaaagtta ccaaaggtgg tccgctgccg 2221 ttcgcttggg acatcctgtc cccgcagttc cagtacggtt ccaaagctta cgttaaacac 2281 ccggctgaca tcccggacta cctgaaactg tccttcccgg aaggtttcaa atgggaacgt 2341 gttatgaact tcgaagacgg tggtgttgtt accgttaccc aggactcctc cctgcaagac 2401 ggtgagttca tctacaaagt taaactgcgt ggtaccaact tcccgtccga cggtccggtt 2461 atgcagaaaa aaaccatggg ttgggaagct tccaccgaac gtatgtaccc ggaagacggt 2521 gctctgaaag gtgaaatcaa aatgcgtctg aaactgaaag acggtggtca ctacgacgct 2581 gaagttaaaa ccacctacat ggctaaaaaa ccggttcagc tgccgggtgc ttacaaaacc 2641 gacatcaaac tggacatcac ctcccacaac gaagactaca ccatcgttga acagtacgaa 2701 cgtgctgaag gtcgtcactc caccggtgct taataacgct gatagtgcta gtgtagatcg 2761 ctactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt 2821 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg 2881 ggcctttctg cgtttatata ctagtagcgg ccgctgcagg gtctctactg cgagacgaa //