LOCUS       .                       5924 bp    DNA     linear   UNK 01-JAN-1980
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_feature    5849..5907
                     /note="multiple cloning site"
                     /note="color: #99ccff"
                     /label="MCS"
     RBS             492..497
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
                     /label="RBS"
     CDS             complement(157..483)
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /transl_table=1
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
                     /label="TrxA"
                     /note="color: #cc99b2"
     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(5924)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /transl_table=1
                     /label="Factor Xa site"
                     /note="color: #cc99b2"
     CDS             complement(118..135)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     CDS             complement(91..108)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /transl_table=1
                     /translation="LVPRGS"
                     /label="thrombin site"
                     /note="color: #cc99b2"
     CDS             complement(5831..5848)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
                     /note="color: #cc99b2"
     promoter        complement(553..571)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff"
                     /label="T7 promoter"
     protein_bind    528..552
                     /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      5717..5764
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
                     /note="color: #ffffff"
                     /label="T7 terminator"
     promoter        884..961
                     /gene="lacI"
                     /note=""
                     /note="color: #ffffff"
                     /label="lacI promoter"
     promoter        complement(5094..5198)
                     /gene="bla"
                     /label="AmpR promoter"
                     /note="color: #ffffff"
     CDS             2853..3044
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /transl_table=1
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL*"
                     /label="rop"
                     /note="color: #993366"
     rep_origin      complement(5225..5680)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
                     /label="f1 ori"
     rep_origin      complement(3474..4062)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /note="color: #ffff00"
                     /label="ori"
     CDS             complement(4233..5093)
                     /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             962..2044
                     /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 ggccagaacc agaaccggcc aggttagcgt cgaggaactc
      181 tttcaactga cctttagaca gtgcacccac tttggttgcc gccacttcac cgtttttgaa
      241 cagcagcaga gtcgggatac cacggatgcc atatttcggc gcagtgccag ggttttgatc
      301 gatgttcagt tttgcaacgg tcagtttgcc ctgatattcg tcagcgattt catccagaat
      361 cggggcgatc attttgcacg gaccgcacca ctctgcccag aaatcgacga ggatcgcccc
      421 gtccgctttg agtacatccg tgtcaaaact gtcgtcagtc aggtgaataa ttttatcgct
      481 catatgtata tctccttctt aaagttaaac aaaattattt ctagagggga attgttatcc
      541 gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc gatctcgatc
      601 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
      661 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
      721 tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc
      781 ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc
      841 ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg
      901 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa
      961 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
     1021 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
     1081 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
     1141 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
     1201 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
     1261 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
     1321 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
     1381 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
     1441 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
     1501 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
     1561 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
     1621 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
     1681 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
     1741 cgggctgcgc gttggtgcgg acatctcggt agtgggatac gacgataccg aagacagctc
     1801 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
     1861 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
     1921 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg
     1981 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     2041 gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga
     2101 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     2161 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     2221 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc
     2281 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
     2341 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg gtgcgcatga
     2401 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
     2461 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
     2521 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
     2581 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg
     2641 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt
     2701 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt
     2761 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca
     2821 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta
     2881 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
     2941 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
     3001 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     3061 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     3121 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     3181 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatatg
     3241 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     3301 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3361 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3421 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3481 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3541 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3601 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3661 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3721 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3781 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3841 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3901 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3961 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4021 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4081 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4141 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4201 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4261 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4321 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4381 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4441 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4501 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     4561 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4621 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4681 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4741 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4801 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4861 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4921 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4981 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5041 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5101 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5161 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5221 cctgaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5281 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     5341 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5401 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5461 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5521 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     5581 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5641 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc aatccggata
     5701 tagttcctcc tttcagcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     5761 ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag
     5821 ccggatctca gtggtggtgg tggtggtgct cgagtgcggc cgcaagcttg tcgacggagc
     5881 tcgaattcgg atccgatatc gccatggcca gaggagagtt agag
//
