LOCUS       .                       4404 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     CDS             413..415
                     /codon_start=1
                     /product="start codon"
                     /transl_table=1
                     /translation="M"
                     /label="ATG"
                     /note="color: #993366"
     RBS             369..373
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
                     /label="RBS"
     misc_feature    346..354
                     /note="translational enhancer from gene 10 of bacteriophage
                     T7 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
                     /label="gene 10 translational enhancer"
     misc_feature    505..554
                     /note="multiple cloning site"
                     /note="color: #99ccff"
                     /label="MCS"
     CDS             3283..4365
                     /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             1074..1934
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="color: #ccffcc"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      2105..2693
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /note="color: #ffff00"
                     /label="ori"
     promoter        982..1073
                     /gene="bla"
                     /label="AmpR promoter"
                     /note="color: #ffffff"
     terminator      757..843
                     /gene="Escherichia coli rrnB"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
                     /note="color: #ffffff"
                     /label="rrnB T1 terminator"
     promoter        3205..3282
                     /gene="lacI (mutant)"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
                     /note="color: #ffffff"
                     /label="lacIq promoter"
     misc_feature    263..332
                     /note="antiterminator from the E. coli rrnG leader region
                     (Berg et al., 1989)"
                     /note="color: #ffffff"
                     /label="rrnG antiterminator"
     CDS             446..478
                     /codon_start=1
                     /note="promotes efficient translation in E. coli"
                     /note="color: #cc99b2"
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /transl_table=1
                     /translation="MASMTGGQQMG"
                     /label="T7 tag (gene 10 leader)"
     misc_feature    193..222
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
                     /note="color: #ffffff"
                     /label="trc promoter"
     terminator      935..962
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
                     /note="color: #ffffff"
                     /label="rrnB T2 terminator"
     CDS             383..409
                     /codon_start=1
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /note="color: #993366"
                     /product="synthetic cistron containing a ribosome binding
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /transl_table=1
                     /translation="MYRLNKEE*"
                     /label="minicistron"
     CDS             482..505
                     /codon_start=1
                     /product="Xpress™ epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /transl_table=1
                     /translation="DLY,DDDDK"
                     /label="Xpress™ tag"
                     /note="color: #cc99b2"
     CDS             425..442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     protein_bind    230..246
                     /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"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
      301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
      361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatgggggg
      421 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      481 ggatctgtac gacgatgacg ataaggatcc gagctcgaga tctgcagctg gtaccatatg
      541 ggaattcgaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
      601 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
      661 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
      721 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
      781 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
      841 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
      901 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
      961 tttgcgtttc tacaaactct ttttgtttat ttttctaaat acattcaaat atgtatccgc
     1021 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1081 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1141 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1201 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1261 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     1321 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1381 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1441 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1501 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1561 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1621 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1681 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1741 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1801 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1861 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1921 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1981 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2041 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2101 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2161 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2221 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     2281 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2341 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2401 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2461 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2521 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2581 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2641 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2701 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2761 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2821 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2881 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     2941 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     3001 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3061 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3121 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa
     3181 ggcgaagcgg catgcattta cgttgacacc atcgaatggt gcaaaacctt tcgcggtatg
     3241 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
     3301 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
     3361 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
     3421 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
     3481 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     3541 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     3601 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     3661 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     3721 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     3781 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     3841 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     3901 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     3961 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     4021 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     4081 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta
     4141 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     4201 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     4261 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4321 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4381 tgtgagttag cgcgaattga tctg
//
