LOCUS       pMCSG81.        4680 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Compact bacterial vector encoding tRNA genes for rare Arg and Ile 
            codons, for ligation-independent cloning (LIC) and addition of a 
            C-terminal 6xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    pMCSG81
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4680)
  AUTHORS   Midwest Center for Structural Genomics
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4680)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     For LIC, amplify the vector with the F and R primers and treat with 
            T4 DNA polymerase plus dCTP.
FEATURES             Location/Qualifiers
     source          1..4680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             complement(158..160)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(169..189)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(205..229)
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(230..248)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     tRNA            complement(523..599)
                     /label=argU
     tRNA            complement(869..944)
                     /label=ileX
     promoter        1084..1161
                     /label=lacI promoter
     CDS             1162..2241
                     /label=lacI
                     /note="lac repressor"
     protein_bind    2257..2278
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2673..3261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3435..4292)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4293..4396)
                     /label=AmpR promoter
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcaa tggtgatggt gatgatgcat tatctttact ccttcttaaa
      181 gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag
      241 tcgtattaat ttcgcgggat cgagatcgat ctcgatcctc tacgccggac gcatcgtggc
      301 cggcatcacc ggtggattca gtcaggcgtc ccattatcag tgcttcagga aatgggcgga
      361 tgggagtcca tagaaatggt tcgtaggtat gctcaccttg cgcctaatca tttgacagag
      421 catgcgagga aaatagacga catttttggt gataatgtcc caaatatgtc ccactctgga
      481 attatggagg atataaagaa ggcgtaactg attgaattgt aatggcgcgc cctgcaggat
      541 tcgaacctgc ggcccacgac ttagaaggtc gttgctctat ccaactgagc taagggcgcg
      601 ttgataccgc aatgcggtgt aatcgcgtga attatacggt caacccttgc tgagtcaatg
      661 gcttttgatc tcgccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
      721 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
      781 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc aaaaaggctg
      841 acgatttctc gtcagccttt gctatatctg gtggcccctg ctggacttga accagcgacc
      901 aagcgattat gagtcgcctg ctctaaccac tgagctaagg ggccatggta gcggattata
      961 aagtaactcc gtgtcgcaat ccgcatgcac cattccttgc ggcggcggtg ctcaacggcc
     1021 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc
     1081 ccggacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag
     1141 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
     1201 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
     1261 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
     1321 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
     1381 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     1441 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     1501 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     1561 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     1621 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     1681 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     1741 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     1801 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     1861 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     1921 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     1981 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     2041 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     2101 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     2161 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     2221 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
     2281 ggcaccggga tctcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     2341 ggcgcggggc atgactacag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     2401 ggcatcagag cagattgtac tgagagtgca ccatatatgc ggtgtgaaat accgcacaga
     2461 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     2521 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2581 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2641 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2701 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2761 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2821 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2881 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2941 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3001 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3061 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     3121 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3181 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3241 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3301 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3361 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3421 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3481 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3541 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3601 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3661 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3721 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     3781 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3841 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3901 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3961 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4021 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4081 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4141 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4201 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4261 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaaat tattgaagca
     4321 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4381 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatgt
     4441 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4501 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4561 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4621 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
