LOCUS       40924_37033        3343 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRhamhIL-10LT, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3343)
  AUTHORS   Guggenbichler F, Buttner C, Rudolph W, Zimmermann K, Gunzer F, 
            Pohlmann C.
  TITLE     Design of a covalently linked human interleukin-10 fusion protein 
            and its secretory expression in Escherichia coli
  JOURNAL   Appl. Microbiol. Biotechnol. (2016) In press
  PUBMED    27430741
REFERENCE   2  (bases 1 to 3343)
  AUTHORS   Auerbach C, Guggenbichler F, Buettner C, Rudolph W, Zimmermann K, 
            Gunzer F, Poehlmann C.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JAN-2016) Institute of Medical Microbiology and 
            Hygiene, TU Dresden, Fetscherstrasse 74, Dresden, Saxony 01307, 
            Germany
REFERENCE   3  (bases 1 to 3343)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3343)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol. (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-JAN-2016) Institute of Medical Microbiology and Hygiene, TU 
            Dresden, Fetscherstrasse 74, Dresden, Saxony 01307, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..119
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     RBS             153..175
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1254..1271
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1303..1350
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1448..1477
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1499..2086
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(2098..2125)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2152..2340)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2344..3156)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(3157..3247)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     terminator      complement(3248..3279)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
ORIGIN
        1 caccacaatt cagcaaattg tgaacatcat cacgttcatc tttccctggt tgccaatggc
       61 ccattttcct gtcagtaacg agaaggtcgc gaattcaggc gctttttaga ctggtcgtag
      121 ggagaccaca acggtttccc tctagaaata attttgttta actataagaa ggagatatac
      181 atatgatgaa gcgcaatatt ctggcagtga tcgtccctgc tctgttagta gcaggtactg
      241 caaacgctag cccaggccaa ggtactcagt ctgaaaactc ctgtacccac tttccgggca
      301 acctgccgaa catgctgcgt gacctgcgtg atgcgtttag ccgtgttaag accttcttcc
      361 agatgaaaga ccagctggat aatctgctgc tgaaagaatc tctgctggaa gatttcaaag
      421 gttatctggg ctgccaggca ctgtccgaaa tgatccagtt ctacctggag gaagtaatgc
      481 cgcaggcgga gaaccaggac ccggatatca aagctcatgt taactccctg ggtgaaaacc
      541 tgaaaacgct gcgcctgcgc ctgcgtcgtt gtcaccgctt cctgccttgc gaaaacaaat
      601 ctaaagcagt ggaacaagtc aagaacgctt tcaacaaact gcaggaaaaa ggtatctata
      661 aagcgatgtc tgagtttgac atcttcatta attacattga agcctacatg accatgaaga
      721 tccgtaacgg tggtggtggt agtggtggcg gtggttcagg tggtggtggc agcagcccag
      781 gccaaggtac tcagtctgaa aactcctgta cccactttcc gggcaacctg ccgaacatgc
      841 tgcgtgacct gcgtgatgcg tttagccgtg ttaagacctt cttccagatg aaagaccagc
      901 tggataatct gctgctgaaa gaatctctgc tggaagattt caaaggttat ctgggctgcc
      961 aggcactgtc cgaaatgatc cagttctacc tggaggaagt aatgccgcag gcggagaacc
     1021 aggacccgga tatcaaagct catgttaact ccctgggtga aaacctgaaa acgctgcgcc
     1081 tgcgcctgcg tcgttgtcac cgcttcctgc cttgcgaaaa caaatctaaa gcagtggaac
     1141 aagtcaagaa cgctttcaac aaactgcagg aaaaaggtat ctataaagcg atgtctgagt
     1201 ttgacatctt cattaattac attgaagcct acatgaccat gaagatccgt aaccatcatc
     1261 accaccatca ctaatagagc ggccgccacc gctgagcaat aactagcata accccttggg
     1321 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cgggtaacga
     1381 attcaagctt gatatcattc aggacgagcc tcagactcca gcgtaactgg actgcaatca
     1441 actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt
     1501 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1561 cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa ctggcttcag
     1621 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1681 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1741 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1801 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1861 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     1921 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     1981 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2041 gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     2101 agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc agaggttttc
     2161 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     2221 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     2281 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcat
     2341 ttagaaaaac tcatcgagca tcaagtgaaa ctgcaattta ttcatatcag gattatcaat
     2401 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     2461 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     2521 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     2581 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
     2641 gccattacgc tcgtcatcaa aatcactcgc accaaccaaa ccgttattca ttcgtgattg
     2701 cgcctgagcg agacgaaata cgcgatcgcc gttaaaagga caattacaaa caggaatcga
     2761 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     2821 ttcttctaat acctggaatg ctgttttccc tgggatcgca gtggtgagta accatgcatc
     2881 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     2941 tagcctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     3001 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     3061 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     3121 cctcgagcaa gacgtttccc gttgaatatg gctcatagct cctgaaaatc tcgataactc
     3181 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     3241 ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta
     3301 tgatttaaat ggtcagtatt gagcgatatc tagagaattc gtc
//
