LOCUS       Exported                6342 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCOLA-AraC-pBAD-UmuD-SP6RNAP, complete sequence.
ACCESSION   KM229376
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6342)
  AUTHORS   Schaerli Y, Munteanu A, Gili M, Cotterell J, Sharpe J, Isalan M.
  TITLE     A unified design space of synthetic stripe-forming networks
  JOURNAL   Nat Commun 5, 4905 (2014)
  PUBMED    25247316
REFERENCE   2  (bases 1 to 6342)
  AUTHORS   Schaerli Y, Munteanu A, Gili M, Cotterell J, Sharpe J, Isalan M.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JUL-2014) Centre for Genomic Regulation (CRG), 
            EMBL/CRG Systems Biology Research Unit, Dr. Aiguader 88, Barcelona, 
            Barcelona 08003, Spain
REFERENCE   3  (bases 1 to 6342)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6342
                     /organism="Cloning vector pCOLA-AraC-pBAD-UmuD-SP6RNAP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1531946"
     RBS             126..148
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             156..2894
                     /codon_start=1
                     /gene="umuD-SP6 RNAP"
                     /product="UmuD-SP6 RNA polymerase"
                     /label=umuD-SP6 RNAP
                     /note="SP6 RNAP with N-terminal degradation tag (UmuD)"
                     /protein_id="AIO10992.1"
                     /translation="MLFIKPADLREIVTFPLFSDLVQCGFPSPGGGSGGGSHMQDLHAI
                     QLQLEEEMFNGGIRRFEADQQRQIAAGSESDTAWNRRLLSELIAPMAEGIQAYKEEYEG
                     KKGRAPRALAFLQCVENEVAAYITMKVVMDMLNTDATLQAIAMSVAERIEDQVRFSKLE
                     GHAAKYFEKVKKSLKASRTKSYRHAHNVAVVAEKSVAEKDADFDRWEAWPKETQLQIGT
                     TLLEILEGSVFYNGEPVFMRAMRTYGGKTIYYLQTSESVGQWISAFKEHVAQLSPAYAP
                     CVIPPRPWRTPFNGGFHTEKVASRIRLVKGNREHVRKLTQKQMPKVYKAINALQNTQWQ
                     INKDVLAVIEEVIRLDLGYGVPSFKPLIDKENKPANPVPVEFQHLRGRELKEMLSPEQW
                     QQFINWKGECARLYTAETKRGSKSAAVVRMVGQARKYSAFESIYFVYAMDSRSRVYVQS
                     STLSPQSNDLGKALLRFTEGRPVNGVEALKWFCINGANLWGWDKKTFDVRVSNVLDEEF
                     QDMCRDIAADPLTFTQWAKADAPYEFLAWCFEYAQYLDLVDEGRADEFRTHLPVHQDGS
                     CSGIQHYSAMLRDEVGAKAVNLKPSDAPQDIYGAVAQVVIKKNALYMDADDATTFTSGS
                     VTLSGTELRAMASAWDSIGITRSLTKKPVMTLPYGSTRLTCRESVIDYIVDLEEKEAQK
                     AVAEGRTANKVHPFEDDRQDYLTPGAAYNYMTALIWPSISEVVKAPIVAMKMIRQLARF
                     AAKRNEGLMYTLPTGFILEQKIMATEMLRVRTCLMGDIKMSLQVETDIVDEAAMMGAAA
                     PNFVHGHDASHLILTVCELVDKGVTSIAVIHDSFGTHADNTLTLRVALKGQMVAMYIDG
                     NALQKLLEEHEERWMVDTGIEVPEQGEFDLNEIMDSEYVFA"
     gene            156..2894
                     /gene="umuD-SP6 RNAP"
                     /label=umuD-SP6 RNAP
     regulatory      2990..3242
                     /regulatory_class="terminator"
     terminator      2990..3037
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3046..3132
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3224..3251
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3439..4254)
                     /codon_start=1
                     /gene="kanR"
                     /product="kanamycin resistance protein"
                     /label=kanR
                     /note="confers resistance to kanamycin"
                     /protein_id="AIO10994.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(3439..4254)
                     /gene="kanR"
                     /label=kanR
     promoter        complement(4255..4346)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4364..4999)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     CDS             complement(5149..6027)
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /note="DNA-binding transcriptional regulator"
                     /protein_id="AIO10993.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            complement(5149..6027)
                     /gene="araC"
                     /label=araC
     promoter        6055..6339
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     regulatory      6302..6330
                     /regulatory_class="promoter"
                     /note="pBAD promoter"