LOCUS       .                       5915 bp    DNA     linear   UNK 01-JAN-1980
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     RBS             484..489
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
                     /label="RBS"
     CDS             complement(149..475)
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /transl_table=1
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
                     /label="TrxA"
                     /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(110..127)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     CDS             complement(83..100)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /transl_table=1
                     /translation="LVPRGS"
                     /label="thrombin site"
                     /note="color: #cc99b2"
     CDS             complement(5823..5840)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     promoter        complement(545..563)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff"
                     /label="T7 promoter"
     protein_bind    520..544
                     /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(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"
     terminator      5709..5756
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
                     /note="color: #ffffff"
                     /label="T7 terminator"
     promoter        876..953
                     /gene="lacI"
                     /note=""
                     /note="color: #ffffff"
                     /label="lacI promoter"
     promoter        complement(5086..5190)
                     /gene="bla"
                     /label="AmpR promoter"
                     /note="color: #ffffff"
     CDS             2845..3036
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /transl_table=1
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL*"
                     /label="rop"
                     /note="color: #993366"
     rep_origin      complement(5217..5672)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
                     /label="f1 ori"
     rep_origin      complement(3466..4054)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /note="color: #ffff00"
                     /label="ori"
     CDS             complement(4225..5085)
                     /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
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     CDS             954..2036
                     /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"
     misc_feature    5841..5899
                     /note="multiple cloning site"
                     /note="color: #99ccff"
                     /label="MCS"
ORIGIN
        1 tcttgtcgtc gtcatcggta cccagatctg ggctgtccat gtgctggcgt tcgaatttag
       61 cagcagcggt ttctttcata ccagaaccgc gtggcaccag accagaagaa tgatgatgat
      121 gatggtgcat atggccagaa ccagaaccgg ccaggttagc gtcgaggaac tctttcaact
      181 gacctttaga cagtgcaccc actttggttg ccgccacttc accgtttttg aacagcagca
      241 gagtcgggat accacggatg ccatatttcg gcgcagtgcc agggttttga tcgatgttca
      301 gttttgcaac ggtcagtttg ccctgatatt cgtcagcgat ttcatccaga atcggggcga
      361 tcattttgca cggaccgcac cactctgccc agaaatcgac gaggatcgcc ccgtccgctt
      421 tgagtacatc cgtgtcaaaa ctgtcgtcag tcaggtgaat aattttatcg ctcatatgta
      481 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa
      541 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga tcctctacgc
      601 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
      661 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
      721 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
      781 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
      841 gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct
      901 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
      961 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     1021 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     1081 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     1141 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     1201 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     1261 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     1321 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     1381 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     1441 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     1501 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     1561 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     1621 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     1681 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     1741 gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     1801 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     1861 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     1921 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     1981 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     2041 aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg
     2101 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     2161 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc
     2221 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta
     2281 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc
     2341 ggcgagaagc aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc
     2401 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
     2461 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
     2521 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
     2581 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
     2641 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
     2701 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
     2761 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
     2821 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
     2881 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
     2941 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc
     3001 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3061 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3121 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     3181 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatata tgcggtgtga
     3241 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     3301 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3361 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3421 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3661 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3721 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3901 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4021 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4081 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4141 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4201 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4261 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4321 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4381 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4441 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4501 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg
     4561 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4621 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4681 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4741 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4801 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4861 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4921 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4981 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5041 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5101 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5161 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat
     5221 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     5281 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     5341 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     5401 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     5461 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     5521 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     5581 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     5641 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccaatccgga tatagttcct
     5701 cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat
     5761 tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc agccggatct
     5821 cagtggtggt ggtggtggtg ctcgagtgcg gccgcaagct tgtcgacgga gctcgaattc
     5881 ggatccgata tcgccatggt tgaggagaag cccgg
//
