LOCUS       .                       4541 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_feature    192..202
                     /note="stop codons in all three reading frames"
                     /note="color: #ff0000"
                     /label="stop codons"
     misc_feature    145..192
                     /note="multiple cloning site"
                     /note="color: #99ccff"
                     /label="MCS"
     CDS             complement(1249..2331)
                     /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(3622..4437)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /note="confers resistance to kanamycin"
                     /note="color: #ccffcc"
                     /product="aminoglycoside phosphotransferase"
                     /transl_table=1
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
                     /label="KanR"
     CDS             346..1005
                     /codon_start=1
                     /gene="cat"
                     /note="confers resistance to chloramphenicol"
                     /note="color: #ccffcc"
                     /product="chloramphenicol acetyltransferase"
                     /transl_table=1
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA*"
                     /label="CmR"
     rep_origin      complement(2927..3515)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /note="color: #ffff00"
                     /label="ori"
     terminator      208..302
                     /gene=""
                     /note="transcription terminator from phage lambda"
                     /note="color: #ffffff"
                     /label="lambda t0 terminator"
     terminator      1070..1156
                     /gene="Escherichia coli rrnB"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
                     /note="color: #ffffff"
                     /label="rrnB T1 terminator"
     promoter        complement(2332..2409)
                     /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"
     promoter        10..54
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
                     /note="color: #ffffff"
                     /label="T5 promoter"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     protein_bind    30..46
                     /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"
     protein_bind    62..78
                     /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"
     CDS             115..117
                     /codon_start=1
                     /product="start codon"
                     /transl_table=1
                     /translation="M"
                     /label="ATG"
                     /note="color: #993366"
     RBS             101..106
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
                     /label="RBS"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatcgcatc accatcacca tcacggatcc gcatgcgagc tcggtacccc gggtcgacct
      181 gcagccaagc ttaattagct gagcttggac tcctgttgat agatccagta atgacctcag
      241 aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga
      301 atccaagcta gcttggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
      361 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
      421 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
      481 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
      541 ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg
      601 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
      661 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
      721 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
      781 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
      841 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
      901 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
      961 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt
     1021 tattggtgcc cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg
     1081 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     1141 cctgagtagg acaaatccgc cctctagatt acgtgcagtc gatgataagc tgtcaaacat
     1201 gagaattgtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     1261 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1321 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     1381 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     1441 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     1501 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     1561 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     1621 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     1681 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     1741 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     1801 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     1861 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     1921 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     1981 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     2041 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     2101 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     2161 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     2221 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     2281 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     2341 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcaccattc
     2401 gatggtgtcg gaatttcggg cagcgttggg tcctggccac gggtgcgcat gatctagagc
     2461 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     2521 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     2581 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     2641 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     2701 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     2761 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2821 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2881 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2941 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3001 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3061 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3121 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3181 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3241 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3301 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3361 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3421 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3481 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3541 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg gcgttaaggg
     3601 attttggtca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     3661 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     3721 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     3781 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     3841 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     3901 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     3961 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     4021 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     4081 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     4141 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     4201 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     4261 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     4321 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     4381 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
     4441 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc atgacattaa
     4501 cctataaaaa taggcgtatc acgaggccct ttcgtcttca c
//
