LOCUS       .                       5446 bp    DNA     linear   UNK 01-JAN-1980
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_feature    5371..5432
                     /note="multiple cloning site"
                     /note="color: #99ccff"
                     /label="MCS"
     CDS             complement(1..9)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /transl_table=1
                     /translation="IEG"
                     /label="Factor Xa site"
                     /note="color: #cc99b2"
     CDS             complement(5446)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /transl_table=1
                     /label="Factor Xa site"
                     /note="color: #cc99b2"
     RBS             147..152
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
                     /label="RBS"
     CDS             complement(136..138)
                     /codon_start=1
                     /product="start codon"
                     /transl_table=1
                     /translation="M"
                     /label="ATG"
                     /note="color: #993366"
     CDS             complement(118..135)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     CDS             complement(5353..5370)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     promoter        complement(208..226)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff"
                     /label="T7 promoter"
     protein_bind    183..207
                     /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             complement(40..84)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /transl_table=1
                     /translation="KETAAAKFERQHMDS"
                     /label="S-Tag"
                     /note="color: #cc99b2"
     terminator      5239..5286
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
                     /note="color: #ffffff"
                     /label="T7 terminator"
     promoter        539..616
                     /gene="lacI"
                     /note=""
                     /note="color: #ffffff"
                     /label="lacI promoter"
     CDS             2508..2699
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /transl_table=1
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL*"
                     /label="rop"
                     /note="color: #993366"
     rep_origin      complement(4747..5202)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
                     /label="f1 ori"
     rep_origin      complement(3129..3717)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /note="color: #ffff00"
                     /label="ori"
     CDS             3839..4654
                     /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             617..1699
                     /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"
ORIGIN
        1 accctcaata ccggagccac caccggtacc cagatctggg ctgtccatgt gctggcgttc
       61 gaatttagca gcagcggttt ctttcatacc agaaccgcgt ggcaccagac cagaagaatg
      121 atgatgatga tggtgcatat gtatatctcc ttcttaaagt taaacaaaat tatttctaga
      181 ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg
      241 agatcgatct cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
      301 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
      361 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
      421 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
      481 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga
      541 caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
      601 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
      661 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
      721 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
      781 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
      841 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
      901 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
      961 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     1021 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     1081 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
     1141 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     1201 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
     1261 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
     1321 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
     1381 cgccattacc gagtccgggc tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga
     1441 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     1501 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     1561 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
     1621 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     1681 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
     1741 cgggatctcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     1801 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     1861 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga
     1921 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca
     1981 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccc
     2041 cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc
     2101 ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca
     2161 aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc
     2221 tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct
     2281 gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag
     2341 tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca
     2401 gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg
     2461 gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg
     2521 aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga
     2581 aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg
     2641 ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2701 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2761 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     2821 gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     2881 agtgcaccat atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     2941 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3001 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3061 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3121 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3181 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3241 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3301 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3361 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3421 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3481 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3541 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     3601 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3661 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3721 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3781 attttggtca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     3841 gagccatatt caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc
     3901 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     3961 tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     4021 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     4081 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     4141 ccccgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     4201 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     4261 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     4321 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     4381 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     4441 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     4501 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     4561 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     4621 gcagtttcat ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat
     4681 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4741 cacctgaaat tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4801 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4861 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4921 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4981 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     5041 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     5101 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     5161 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccaatccgga
     5221 tatagttcct cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta
     5281 tgctagttat tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc
     5341 agccggatct cagtggtggt ggtggtggtg ctcgagtgcg gccgcaagct tgtcgacgga
     5401 gctcgaattc ggatccgata tcgccatggc cagaggagag ttagag
//
