LOCUS       40924_6566        8005 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle expression vector pBJ2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8005)
  AUTHORS   Equbal MJ, Agarwal GP.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   2  (bases 1 to 8005)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8005)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (01-JUN-2010) Department of Biochemical Engineering "
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8005
                     /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"
     misc_feature    complement(158..203)
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             complement(207..239)
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             complement(243..260)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(270..287)
                     /label=6xHis
                     /note="6xHis affinity tag"
     RBS             complement(306..328)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(343..367)
                     /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(368..386)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             461..1117
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        1672..1749
                     /label=lacI promoter
     misc_feature    2701..5451
                     /note="derived from BL1 replicon of Corynebacterium
                     glutamicum plasmid pBL1 INSD accession X03987; involved in 
                     plasmid pBJ1 replication in Corynebacterium glutamicum"
     misc_feature    5593..5735
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5921..6509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             6631..7443
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(7539..7994)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgcgaccc atttgctgtc caccagtcat gctagccata
      241 tggctgccgc gcggcaccag gccgctgctg tgatgatgat gatgatggct gctgcccatg
      301 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
      361 caattcccct atagtgagtc gtattaattt cgcgggatcg agatcactac cgggcgtatt
      421 ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
      481 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
      541 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
      601 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
      661 gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag
      721 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
      781 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
      841 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
      901 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
      961 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     1021 ggcgattcag gttcatcatg ccgtctgtga tggcttccat gtcggcagaa tgcttaatga
     1081 attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg
     1141 gtgcccttaa acgcctggtg ctacgcctga ataagtgata ataagcggat gaatggcaga
     1201 aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt
     1261 atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct
     1321 caaatgcctg aggccagttt gctcaggctc tccccgtgga ggtaataatt gacgatatga
     1381 tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
     1441 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
     1501 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg
     1561 ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac
     1621 tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc
     1681 gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     1741 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
     1801 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
     1861 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     1921 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     1981 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     2041 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     2101 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     2161 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     2221 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
     2281 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
     2341 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
     2401 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
     2461 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
     2521 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
     2581 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
     2641 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     2701 aacaacaaga cccatcatag tttgcccccg cgacattgac cataaattca tcgcacaaaa
     2761 tatcgaacgg ggtttatgcc gcttttagtg ggtgcgaaga atagtctgct cattacccgc
     2821 gaacaccgcc gcattcagat cacgcttagt agcgtcccca tgagtaggca gaaccgcgtc
     2881 caagtccaca tcatccataa cgatcatgca cggggtggaa tccacaccca gacttgccag
     2941 cacctcatta gcgacacgtt gcgcagcggc cacgtcctta gccttatcca cgcaatctag
     3001 aacgtactgc ctaaccgcga aatcagactg aatcagtttc caatcatcgg gcttcaccaa
     3061 agcaacagca acgcgggttg attcgacccg ttccggtgct tccagaccgg cgagcttgta
     3121 cagttcttct tccatttcac gacgtacatc agcgtctatg taatcaatgc ccaaagcacg
     3181 cttagcccca cgtgaccagg acgaacgcag gtttttagaa ccaacctcat actcacgcca
     3241 ccgagccacc aaaacagcgt ccatatcctc gccggcgtcg ctttgatcgg ccaacatatc
     3301 caacatctga aacggcgtgt acgacccctt agacgcggtt ttagtagcgg agccagtcag
     3361 ttcctgagac atgcccttag cgaggtaggt tgccattttc gcagcgtctc caccccaggt
     3421 agacacctga tcaagtttga ccccgtgctc acgcagtggc gcgtccatac cggccttaac
     3481 cacaccagca gaccagcggg aaaacatgga atcctcaaac gccttgagtt catcgtcaga
     3541 cagtggacga tccaagaaca acagcatgtt gcggtgcaag tgccaaccgt tcgcccaaga
     3601 gtctgtgacc tcatagtcac tataggtgtg ctccaccccg taccgtgcac gttctttctt
     3661 ccactgagat gttttcacca tcgaagagta cgcagtctta atacccgctt caacctgcgc
     3721 aaatgactgt gagcggttgt gtcgaacagt gcccacaaac atcatgagcg cgccacccgc
     3781 cgccaagtga ttcttagtag caatagccag ctcaatgcgg cgttcgccca tgacttccaa
     3841 ttcagccaga ggtgaccccc agcgagagtg agagttttgc agaccctcaa actgcgaagc
     3901 accgttagac gaccaggaca ccgcaacagc ttcgtccctg cgccacctat ggcaccccgc
     3961 cagagcctta ctattggtga tcttgtacat gacgttttgc ctacgccacg ccctagcgcg
     4021 agtgacctta gaaccctcat tgacctgcgg ttccttagag gtgttcactt ctatttcagt
     4081 gttactcagt gttacctaga cccgatgttg tgcggggttg cgcagtgcga gtttgtgcgg
     4141 gtgttgtgcc cgttgtctta gctagtgcta tggttgtcaa ttgaaacccc ttcgggttat
     4201 gtggcccccg tgcatatgag ttggtagctc gcacgggggt ttgtcttgtc tagggactat
     4261 taatttttag tggtgtttgg tggccgccta gcttggctat gcgtgccagc ttacccgtac
     4321 tcaatgttaa agatttgcat cgacatggga gggttacgtg tccgatacct agggggggta
     4381 tccgcgacta ggtgccccgg tgctcactgt ctgtaccggc ggggcaagcc ccacaccccg
     4441 catggacagg gtggctccgc cccctgcacc cccagcaatc tgcatgtaca tgttttacac
     4501 attagcacga catgactgca tgtgcatgca ctgcatgcag actaggtaaa tatgagtatg
     4561 tacgactagt aacaggagca ctgcacataa tgaatgagtt gcaggacaat gtttgctacg
     4621 catgcgcatg acatatcgca ggaaagctac tagagtctta aagcatggca accaaggcac
     4681 agctagaaca gcaactacaa gaagctcaac aggcactaca ggcgcagcaa gcgcaggcac
     4741 aagccaccat cgaagcacta gaagcgcagg caaaggctaa gcccgtcgtg gtcaccgcac
     4801 gcgttccttt ggcactacgt gaggacatga agcgcgcagg catgcagaac ggtgaaaacc
     4861 tccaagagtt catgatcgcc gcgtttaccg agcggctaga aaagctcacc accaccgaca
     4921 acgaggaaaa caatgtctaa cccactagtt ctctttgccc accgtgaccc ggtaaatgac
     4981 gtgacgttcg agtgcattga gcacgccacc tacgacacac tttcacacgc taaagaccag
     5041 atcaccgccc aaatgcaagc cctagacgaa gaagccgccc tactgcccta atgggtgttt
     5101 catgggtgtt tccctagtgt ttcatggtgt tttcacctaa gctagggaat tgcgcgagaa
     5161 gtctcgcaaa aatcagcaac ccccggaacc acacagttca cgggggttct tctatgccag
     5221 aaatcagaaa ggggaaccag tgaacgaccc cgaatattgg atcacagcgc agcaggtcgc
     5281 cgcccgcgta gctctcaccc cggccaccat taaaaagtgg gcaaacgagg gaaaaatcac
     5341 cgcatacaag atcggcaagt ccgtccgatt caaagcatca gacgtagaca agcttgccac
     5401 accccgagac gcagcctaga agccattcta agcgactatc tagccccggt tctgcctcgc
     5461 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     5521 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     5581 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt
     5641 aactatgcgg catcagagca gattgtactg agagtgcacc atatatgcgg tgtgaaatac
     5701 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     5761 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5821 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5881 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5941 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6001 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6061 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6121 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6181 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6241 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6301 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6361 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6421 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6481 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6541 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgaacaat aaaactgtct
     6601 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc
     6661 tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     6721 gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca
     6781 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     6841 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     6901 cctgatgatg catggttact caccactgcg atccccggga aaacagcatt ccaggtatta
     6961 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     7021 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     7081 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     7141 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     7201 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     7261 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     7321 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     7381 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     7441 ttctaagaat taattcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7501 aggggttccg cgcacatttc cccgaaaagt gccacctgaa attgtaaacg ttaatatttt
     7561 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     7621 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     7681 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     7741 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     7801 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     7861 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     7921 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     7981 gctacagggc gcgtcccatt cgcca
//
