LOCUS       .                       5437 bp    DNA     linear   UNK 01-JAN-1980
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_feature    5363..5421
                     /note="multiple cloning site"
                     /note="color: #99ccff"
                     /label="MCS"
     CDS             609..1691
                     /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             3831..4646
                     /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"
     rep_origin      complement(3121..3709)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /note="color: #ffff00"
                     /label="ori"
     rep_origin      complement(4739..5194)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
                     /label="f1 ori"
     CDS             2500..2691
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /transl_table=1
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL*"
                     /label="rop"
                     /note="color: #993366"
     promoter        531..608
                     /gene="lacI"
                     /note=""
                     /note="color: #ffffff"
                     /label="lacI promoter"
     terminator      5231..5278
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
                     /note="color: #ffffff"
                     /label="T7 terminator"
     CDS             complement(32..76)
                     /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    175..199
                     /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        complement(200..218)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff"
                     /label="T7 promoter"
     CDS             complement(5345..5362)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     CDS             complement(110..127)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     CDS             complement(2..16)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /transl_table=1
                     /translation="DDDDK"
                     /label="enterokinase site"
                     /note="color: #cc99b2"
     CDS             complement(128..130)
                     /codon_start=1
                     /product="start codon"
                     /transl_table=1
                     /translation="M"
                     /label="ATG"
                     /note="color: #993366"
     CDS             complement(83..100)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /transl_table=1
                     /translation="LVPRGS"
                     /label="thrombin site"
                     /note="color: #cc99b2"
     RBS             139..144
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
                     /label="RBS"
ORIGIN
        1 tcttgtcgtc gtcatcggta cccagatctg ggctgtccat gtgctggcgt tcgaatttag
       61 cagcagcggt ttctttcata ccagaaccgc gtggcaccag accagaagaa tgatgatgat
      121 gatggtgcat atgtatatct ccttcttaaa gttaaacaaa attatttcta gaggggaatt
      181 gttatccgct cacaattccc ctatagtgag tcgtattaat ttcgcgggat cgagatcgat
      241 ctcgatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
      301 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat
      361 gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc
      421 catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact
      481 gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgagatcccg gacaccatcg
      541 aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
      601 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
      661 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
      721 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
      781 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
      841 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
      901 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
      961 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
     1021 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
     1081 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
     1141 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
     1201 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
     1261 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
     1321 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
     1381 ccgagtccgg gctgcgcgtt ggtgcggaca tctcggtagt gggatacgac gataccgaag
     1441 acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
     1501 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
     1561 tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct
     1621 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     1681 gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc accgggatct
     1741 cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg
     1801 actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg
     1861 gcagcgctct gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc
     1921 ctgtcgcttg cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc
     1981 gccaccaaac gtttcggcga gaagcaggcc attatcgccg gcatggcggc cccacgggtg
     2041 cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt
     2101 tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct
     2161 gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg
     2221 cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta
     2281 ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt
     2341 ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg
     2401 gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt
     2461 acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc
     2521 gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac
     2581 gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag
     2641 ctttaccgca gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2701 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     2761 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     2821 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     2881 atatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     2941 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3001 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3061 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3121 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3181 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3241 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3301 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3361 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3421 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3481 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3541 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3601 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3661 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3721 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3781 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     3841 ttcaacggga aacgtcttgc tctaggccgc gattaaattc caacatggat gctgatttat
     3901 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt
     3961 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     4021 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     4081 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggga
     4141 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     4201 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     4261 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     4321 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     4381 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     4441 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     4501 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     4561 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     4621 atttgatgct cgatgagttt ttctaagaat taattcatga gcggatacat atttgaatgt
     4681 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgaa
     4741 attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4801 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4861 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4921 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4981 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     5041 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     5101 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     5161 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccaatccg gatatagttc
     5221 ctcctttcag caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctagtt
     5281 attgctcagc ggtggcagca gccaactcag cttcctttcg ggctttgtta gcagccggat
     5341 ctcagtggtg gtggtggtgg tgctcgagtg cggccgcaag cttgtcgacg gagctcgaat
     5401 tcggatccga tatcgccatg gttgaggaga agcccgg
//
