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=Ha
                     /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=I
                     /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 acgtctcgat ccagagaccg 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 gggtctctgc
     1141 gtcgagacga 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
//
