LOCUS       pGS-21a.        6169 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expression of 6xHis- and GST-tagged proteins. 
            Two 6xHis sequences are encoded in the vector.
ACCESSION   .
VERSION     .
KEYWORDS    pGS-21a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6169)
  AUTHORS   GenScript
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6169)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             89..91
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             101..118
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             128..781
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     CDS             818..832
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     misc_feature    834..892
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             893..910
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      977..1024
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1061..1516
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1543..1647
                     /label=AmpR promoter
     CDS             1648..2505
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2679..3267
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3453..3595)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3700..3888)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(4663..4684)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4700..5779)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(5780..5857)
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gtctggttct catcatcatc atcatcatag
      121 cagcggtatg tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg
      181 acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg
      241 tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta
      301 tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa
      361 gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc
      421 ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct
      481 caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg
      541 tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc
      601 tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg
      661 ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta
      721 tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa
      781 atcggatctg ggccacacag gccatagatc tggtaccgac gacgacgaca aggccatggc
      841 tgatatcgga tccgaattcg agctcgtcga caagcttgcg gccgcactcg agcaccacca
      901 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
      961 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1021 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     1081 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1141 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1201 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1261 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     1321 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     1381 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1441 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     1501 ttaacgttta caatttcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1561 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1621 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1681 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1741 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1801 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1861 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     1921 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1981 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2041 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2101 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     2161 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     2221 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     2281 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     2341 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     2401 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2461 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2521 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2581 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2641 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2701 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2761 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2821 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2881 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2941 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3001 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3061 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3121 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3181 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3241 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3301 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3361 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3421 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     3481 ctgtgcggta tttcacaccg catatatggt gcactctcag tacaatctgc tctgatgccg
     3541 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     3601 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     3661 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     3721 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     3781 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     3841 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     3901 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     3961 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     4021 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     4081 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     4141 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     4201 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     4261 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     4321 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
     4381 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     4441 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     4501 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     4561 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     4621 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     4681 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4741 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     4801 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     4861 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     4921 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatgt
     4981 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     5041 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     5101 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     5161 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     5221 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     5281 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     5341 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     5401 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     5461 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     5521 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     5581 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     5641 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     5701 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     5761 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     5821 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     5881 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     5941 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     6001 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     6061 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     6121 acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaat
//
