LOCUS       .                       3669 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     RBS             58..63
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
                     /label="RBS"
     CDS             71..73
                     /codon_start=1
                     /product="start codon"
                     /transl_table=1
                     /translation="M"
                     /label="ATG"
                     /note="color: #993366"
     CDS             complement(2447..3529)
                     /codon_start=1
                     /gene="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-β-D-thiogalactopyranoside (IPTG)."
                     /note="color: #993366"
                     /product="lac repressor"
                     /transl_table=1
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ*"
                     /label="lacI"
     CDS             complement(579..1394)
                     /codon_start=1
                     /note="confers resistance to kanamycin in bacteria or G418
                     (Geneticin®) in eukaryotes"
                     /note="color: #ccffcc"
                     /product="aminoglycoside phosphotransferase"
                     /transl_table=1
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
                     /label="KanR"
     rep_origin      complement(1502..2251)
                     /direction=LEFT
                     /note="Plasmids containing the RSF 1030 origin of
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
                     /note="color: #ffff00"
                     /label="RSF ori"
     promoter        complement(1395..1486)
                     /gene="bla"
                     /label="AmpR promoter"
                     /note="color: #ffffff"
     promoter        complement(3530..3607)
                     /gene="lacI"
                     /note=""
                     /note="color: #ffffff"
                     /label="lacI promoter"
     terminator      302..349
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
                     /note="color: #ffffff"
                     /label="T7 terminator"
     CDS             206..250
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /transl_table=1
                     /translation="KETAAAKFERQHMDS"
                     /label="S-Tag"
                     /note="color: #cc99b2"
     protein_bind    3..27
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
                     /label="lac operator"
     promoter        2..3653
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff"
                     /label="T7 promoter"
     CDS             77..94
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     CDS             113..127
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /transl_table=1
                     /translation="DDDDK"
                     /label="enterokinase site"
                     /note="color: #cc99b2"
     misc_feature    69..278
                     /note="multiple cloning site"
                     /note="color: #99ccff"
                     /label="MCS"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggcacatc accaccacca tcacgtgggt accggttcga atgatgacga
      121 cgacaagagt ccggatccca attgggagct cgtgtacacg gcgcgcctgc aggtcgacaa
      181 gcttgcggcc gcactcgagt ctggtaaaga aaccgctgct gcgaaatttg aacgccagca
      241 catggactcg tctactagcg cagcttaatt aacctaggct gctgccaccg ctgagcaata
      301 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca
      361 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
      421 atagacataa gcggctattt aacgaccctg ccctgaaccg acgacaagct gacgaccggg
      481 tctccgcaag tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
      541 aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc
      601 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
      661 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
      721 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
      781 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
      841 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
      901 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
      961 cggtcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     1021 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     1081 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     1141 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     1201 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     1261 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     1321 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
     1381 tgaatatggc tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     1441 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg catgcagcgc
     1501 tcttccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggtg
     1561 tcagctcact caaaagcggt aatacggtta tccacagaat caggggataa agccggaaag
     1621 aacatgtgag caaaaagcaa agcaccggaa gaagccaacg ccgcaggcgt ttttccatag
     1681 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agccagaggt ggcgaaaccc
     1741 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1801 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1861 ttctcatagc tcacgctgtt ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1921 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1981 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccattg gtaactgatt
     2041 tagaggactt tgtcttgaag ttatgcacct gttaaggcta aactgaaaga acagattttg
     2101 gtgagtgcgg tcctccaacc cacttacctt ggttcaaaga gttggtagct cagcgaacct
     2161 tgagaaaacc accgttggta gcggtggttt ttctttattt atgagatgat gaatcaatcg
     2221 gtctatcaag tcaacgaaca gctattccgt tactctagat ttcagtgcaa tttatctctt
     2281 caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat
     2341 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     2401 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     2461 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2521 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     2581 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     2641 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     2701 cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa
     2761 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     2821 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     2881 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     2941 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     3001 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     3061 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     3121 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     3181 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     3241 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     3301 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     3361 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     3421 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     3481 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     3541 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     3601 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa attaatacga
     3661 ctcactata
//
