LOCUS       40924_25481        8703 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression shuttle vector pIMB4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8703)
  AUTHORS   Zhu Y, Wang L, Du Y, Wang S, Yu T, Hong B.
  TITLE     Heterologous expression of human interleukin-6 in Streptomyces 
            lividans TK24 using novel secretory expression vectors
  JOURNAL   Biotechnol. Lett. 33 (2), 253-261 (2011)
  PUBMED    20931350
REFERENCE   2  (bases 1 to 8703)
  AUTHORS   Zhu Y, Wang L, Du Y, Wang S, Yu T, Hong B.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JUL-2010) Key Laboratory of Biotechnology of 
            Antibiotics of Ministry of Health, Institute of Medicinal 
            Biotechnology, Chinese Academy of Medical Sciences and Peking Union 
            Medical College, No. 1 Tiantan Xili, Beijing, Beijing 100050, China
REFERENCE   3  (bases 1 to 8703)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8703)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
            Lett."; date: "2011"; volume: "33"; issue: "2"; pages: "253-261"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-JUL-2010) Key Laboratory of Biotechnology of Antibiotics of 
            Ministry of Health, Institute of Medicinal Biotechnology, Chinese 
            Academy of Medical Sciences and Peking Union Medical College, No. 1 
            Tiantan Xili, Beijing, Beijing 100050, China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             177..977
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
     misc_feature    1452..1481
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    complement(1482..1580)
                     /label=signal peptide of CagA
                     /note="signal peptide of CagA"
     regulatory      complement(1581..1839)
                     /label=cagA promoter
                     /note="cagA promoter"
                     /regulatory_class="promoter"
     regulatory      complement(1846..2057)
                     /label=ermE* promoter
                     /note="ermE* promoter"
                     /regulatory_class="promoter"
     primer_bind     complement(2066..2082)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2112..2130)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2151..2167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2175..2191)
                     /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(2199..2229)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2244..2265)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2553..3141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3315..4172)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4173..4277)
                     /label=AmpR promoter
     rep_origin      complement(4303..4758)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     4900..4916
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4926..4944
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(5303..6739)
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="replication protein of pSGL1 in Streptomyces"
                     /protein_id="ADM21212.1"
                     /translation="MRTRDARWTLREGLQRVTEDKGLKGCGRSAIAGGVGVVLNGKTAH
                     LSGVATCGKIHLCPVCSAKIRAARSEEIDTVTGAWQAAEHGLAMMTLTLRHCRRMPLGT
                     IRRAERGGLIGVQPGHLHIHTIVLWIRARWVRSVEGQGATGPTTVDSGSTSPGRSAQQF
                     SRYLFKSQDGKARFERWAPGAELTSADKAGRKASRMPFEVAAGAAAGLAEDVPLWQSTS
                     TAPMGSGQSTGPTACGHCSASSCKLDERTDEEIASEDVGGTGVALIPAETWYGVILRHR
                     GRSLDLIRGRRDRRRRRGPVAHHRMGAGVGPGRSPGPGDPDRLTLFWPGGSSRPPGRIG
                     LHRICAGRTGSASRNPLRAPGRARAPGARRRRPIRRISWHGRSRPADERDDDGREPVRL
                     VRRAGPTVRDRAQARVLRADASGAGLPGTAAAAGGGRGGCPGARGRSVGIVSCQLSKRG
                     PRNGVPIRQLSKRLRGRSAP"
     gene            complement(5303..6739)
                     /gene="rep"
                     /label=rep
     rep_origin      complement(6740..7139)
                     /direction=LEFT
                     /label=origin of pSGL1
                     /note="origin of pSGL1"
     CDS             complement(8042..8410)
                     /label=traJ
                     /note="oriT-recognizing protein"
     oriT            complement(8443..8552)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
ORIGIN
        1 ctcacggtaa ctgatgccgt atttgcagta ccagcgtacg gcccacagaa tgatgtcacg
       61 ctgaaaatgc cggcctttga atgggttcat gtgcagctcc atcagcaaaa ggggatgata
      121 agtttatcac caccgactat ttgcaacagt gccgttgatc gtgctatgat cgactgatgt
      181 catcagcggt ggagtgcaat gtcgtgcaat acgaatggcg aaaagccgag ctcatcggtc
      241 agcttctcaa ccttggggtt acccccggcg gtgtgctgct ggtccacagc tccttccgta
      301 gcgtccggcc cctcgaagat gggccacttg gactgatcga ggccctgcgt gctgcgctgg
      361 gtccgggagg gacgctcgtc atgccctcgt ggtcaggtct ggacgacgag ccgttcgatc
      421 ctgccacgtc gcccgttaca ccggaccttg gagttgtctc tgacacattc tggcgcctgc
      481 caaatgtaaa gcgcagcgcc catccatttg cctttgcggc agcggggcca caggcagagc
      541 agatcatctc tgatccattg cccctgccac ctcactcgcc tgcaagcccg gtcgcccgtg
      601 tccatgaact cgatgggcag gtacttctcc tcggcgtggg acacgatgcc aacacgacgc
      661 tgcatcttgc cgagttgatg gcaaaggttc cctatggggt gccgagacac tgcaccattc
      721 ttcaggatgg caagttggta cgcgtcgatt atctcgagaa tgaccactgc tgtgagcgct
      781 ttgccttggc ggacaggtgg ctcaaggaga agagccttca gaaggaaggt ccagtcggtc
      841 atgcctttgc tcggttgatc cgctcccgcg acattgtggc gacagccctg ggtcaactgg
      901 gccgagatcc gttgatcttc ctgcatccgc cagaggcggg atgcgaagaa tgcgatgccg
      961 ctcgccagtc gattggctga gctcatgagc ggagaacgag atgacgttgg aggggcaagg
     1021 tcgcgctgat tgctggggca acacgtggag cggatcgggg attgtctttc ttcagctcgc
     1081 tgatgatatg ctgacgctca atgccgtttg gcctccgact aacgaaaatc ccgcatttgg
     1141 acggctgatc cgattggcac ggcggacggc gaatggcgga gcagacgctc gtccgggggc
     1201 aatgagatat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     1261 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     1321 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     1381 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
     1441 ttgacattgg ggaattcaga tctgatatcc cgcggcatat gggcgaaggc gacggactgt
     1501 gcggcggcgg ccaggccgat ggatgcgacg gccacgacgc cgaacctgga agcacggcgg
     1561 gacatgtgac gcagcgacat gcgtaggctc cgattcgagg agggggttga tcactccatg
     1621 aaaggatcac ctcgccggac ggccgcctgc atctccctct gtgctctcgt ggatttccgg
     1681 cacgcactcc cgtcgacggc cgcccgcaga atgcggcaga ccccccgcac ctcctccggc
     1741 cccaccgccg taccggtggg cagcgacacg acccgctcgg tgagcgcctc caccttcggg
     1801 agcggatcgg gcgcgtggcg cgcgaggtcg gaccggtagt ctagatgggg tcctcctgtg
     1861 gagtggttct gtatcctacc aaccggcacg attggcccac aacagcatcg cggtgccacg
     1921 tgtggaccgc gtcggtcaga tcctccccgc acctctcgcc agccgtcaag atggaccgcg
     1981 tgcacctgcg atcgccgatc aaccgcgact agcatcgggc gcaagccgcc actcgaacgg
     2041 acactcgcat ggacgtcaag cttatcgata ccgtcgacct cgaggggggg cccggtaccc
     2101 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     2161 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2221 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2281 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2341 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2401 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2461 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2521 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2581 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2641 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2701 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2761 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2821 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2881 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2941 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     3001 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3061 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3121 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3181 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3241 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3301 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3361 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3421 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3481 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3541 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3601 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3661 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3721 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3781 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3841 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3901 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3961 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4021 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4081 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4141 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4201 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4261 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     4321 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4381 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4441 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4501 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4561 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4621 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4681 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4741 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     4801 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     4861 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc
     4921 gcgcgtaata cgactcacta tagggcgaat tggagctccc gcccgaaccg ccagacctcc
     4981 cgccggatca cgagtctccg cacgacctcc ggcccgcagt agatcgccac cagcgggccg
     5041 tacgtcaccg ccaccacagg ccagagccca agcaattcgc ggcctatcag caccccaccg
     5101 atcagcagtg ccacagccag ggtcagcgcg agactgtctc gctcgctcac gatcacaccc
     5161 ccgtacgtag atcaacacgc accgtagtga tcaccgctga caatgtcgcg ggattaccgg
     5221 ccctcgtccc gctcgacacc cggcggaggc ggcagcagcc gccgccgccg cttcgccggg
     5281 gccgcccgcc acgcggacgg aatcacgggg ctgacctgcc ccggagacgt ttcgacaact
     5341 gacggatcgg gaccccgttt cgcgggcccc gtttcgacaa ctgacaactg acgattccga
     5401 cgctgcggcc gcgagcgcct gggcaaccgc ctcgtccacc gcccgctgct gccgccgtgc
     5461 ccggtaggcc agctcccgat gcgtccgccc gcagtactcg cgcttgcgcc cgatcccgga
     5521 ctgtcggacc ggctcgccgc accaggcgca cgggctccct gccgtcgtcg tctcgctcat
     5581 ccgccggacg cgaccggccg tgccatgaga tacgccgtat cgggcgacga cgacgagcac
     5641 caggagcacg agcacgacca ggagcacgaa gagggttcct cgaggcggat ccggttcggc
     5701 cggcacaaat gcggtgcagg ccgatccggc ccggcggccg tgatgaaccg ccgggccaga
     5761 acagggtcag tcggtcgggg tcgccgggac cgggagaacg tcccggcccc actccagccc
     5821 ccatccggtg atgagcgact ggaccgcgcc gacgccgccg atctcggcgg ccccgaatca
     5881 ggtcgagcga gcggccacgg tgccggagga tcacgccgta ccaggtctct gccgggatca
     5941 gggcgacgcc agtgccgcca acgtcctcgg aggcaatctc ctcgtcggtc cgctcgtcga
     6001 gcttgcacga gcttgccgag cagtgcccgc aggccgttgg accagtagat tgcccggatc
     6061 ccatgggcgc cgtgctcgta ctctgccaca gcggcacgtc ctcggcgagc ccggccgccg
     6121 cccctgccgc cacctcgaac ggcatccggg acgccttacg cccggccttg tcggcactgg
     6181 tcagctccgc gcccggcgcc cagcgctcga accgcgcctt gccgtcctgg gacttgaaca
     6241 ggtagcggct gaactgttgg gcgctgcggc cgggggacgt cgatccggag tccacggtcg
     6301 ttgggccggt agccccctga ccctccacgg agcggaccca acgggcccgg atccacagga
     6361 cgatggtatg aatgtgcagg tgcccgggct gaacgccgat caggccgccc ctctcggccc
     6421 gccggatggt gccgagcggc atccgccggc agtgccggag ggtgagcgtc atcatcgcca
     6481 gcccatgctc tgctgcctgc caggcgcccg tgaccgtgtc gatctcctcg gaccgggcgg
     6541 cccggatctt ggctgagcag accgggcaga ggtggatctt tccgcacgtg gcaacgccgg
     6601 acaggtgggc cgtcttgccg ttcaggacga cgccgacgcc gcccgcgatg gcggaccggc
     6661 cgcagccctt cagccccttg tcctcggtga cccgctggag gccttcccgg agggtccacc
     6721 tggcatcgcg ggtccgcatc ccgggcctgt ccgcttttgc atttccggac acgtccgcag
     6781 cgctctgacc tgcacgggtt gcactttctc cggtagtagc tacgcgcctg tccggatttg
     6841 ctgcggccgg tcgtgcctca gcggtcgtgg tgggcagtgc agtgccccgc tcgatggctc
     6901 gctttgcccc ttccggcagg cgtccgagca gtacacacgc ggccgcttgc ccggcgtccg
     6961 ggcgatgggg gtcccgcagt gacagcgggg tccggcgggg cgaggtaccg ggttacgctg
     7021 atctctcgta cgctcgtcac agagcaaacg tcctcactcg gcatgcgacg ccgattcggg
     7081 ggcggcggcc ccggcctcct acagccgggg tcgtgccatt tcctgggccc catgaggccc
     7141 gttcagatcc ttgcagccgt tggggcattc gttgcccccg gtcctcctga cgcgccgtgt
     7201 ggcggatgtg atcaggagag agctaagatt acatagccat cgcgttgcct gcaatgccat
     7261 cgcgatgttg gtgtgtccag acagccgaaa gccccgcccg gatctccggg cggggctcgg
     7321 ctcgttgctg gtcagctctc gtcgggccag ctccgcaggt agagcgagac cgtaacgctc
     7381 ttgccgttct tcgcggtccg ctcggtgcgg ccgtggtagg tcacttcgcc gtcactgtcg
     7441 ggtcgggccg tcagctcgat gccgaggtga tcggcccacg cggtgagcgc ggccagtcgc
     7501 tccgggcggg tcccgctcgg cggctcgccc tggagctggg cggtggtggc cactcgccag
     7561 gtgatcgggg agaggtcagc ctccgtgatc gtggagagca cgttgatcgc ggcgtactgc
     7621 cagcgtcggc ggtcggtgtc ggtgatgggc atggagtctc tcctcagtca ggcggcgggt
     7681 gcgagtcggg cggcaagctg ggcggctgtc accaggatca gccggacatc ggcggcggct
     7741 tcgggcggga tcggggcccc gaccgattcg gccggcacat tcggcggagg ggtcgggacg
     7801 gggcggcggc ggagggccac cgcgaggacg acgaccgcga ccaggagcga gagcaggtag
     7861 gcgagagcca tggtcatgcc cccgccgccg tcgacctgca ggtccccggg gatcggtctt
     7921 gccttgctcg tcggtgatgt acttcaccag ctccgcgaag tcgctcttct tgatggagcg
     7981 catggggacg tgcttggcaa tcacgcgcac cccccggccg ttttagcggc taaaaaagtc
     8041 atggctctgc cctcgggcgg accacgccca tcatgacctt gccaagctcg tcctgcttct
     8101 cttcgatctt cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt
     8161 cgtcggtgag ccagagtttc agcaggccgc ccaggcggcc caggtcgcca ttgatgcggg
     8221 ccagctcgcg gacgtgctca tagtccacga cgcccgtgat tttgtagccc tggccgacgg
     8281 ccagcaggta ggccgacagg ctcatgccgg ccgccgccgc cttttcctca atcgctcttc
     8341 gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt ggcttggttt
     8401 catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag
     8461 caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc
     8521 tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa
     8581 agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg
     8641 aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaagatc cgtcgacctg
     8701 cag
//
