LOCUS       40924_12635        6342 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCOLA-AraC-pBAD-UmuD-SP6RNAP, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6342)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            5, 4905 (2014)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-JUL-2014) Centre for Genomic Regulation (CRG), EMBL/CRG Systems 
            Biology Research Unit, Dr. Aiguader 88, Barcelona, Barcelona 08003, 
            Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             126..148
                     /label=RBS
                     /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
     terminator      2990..3037
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3046..3132
                     /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(3442..4254)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(4255..4346)
                     /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(5152..6027)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        6055..6339
                     /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)"
ORIGIN
        1 gaattcattt cacccaagga gctcgttgag ttacctgcag cgattgtact agttgcggcc
       61 gcctgcgcac cacctgcagg acctcagcaa tcgcttggac cagctttctt aagggtacca
      121 ataattttgt ttaactttaa gaaggagata tacatatgct gttcatcaaa ccggctgatc
      181 tgcgcgaaat cgtgacgttc ccgctgttct ctgacctggt gcaatgtggc tttccgagtc
      241 cgggcggcgg tagtggcggt ggctcccata tgcaggatct gcacgcgatt cagctgcagc
      301 tggaagaaga aatgtttaac ggtggcattc gtcgcttcga agcggatcag caacgtcaga
      361 tcgcggccgg ttcagaatcg gacaccgcct ggaatcgtcg cctgctgagc gaactgattg
      421 ccccgatggc ggaaggcatt caggcgtata aagaagaata cgaaggtaaa aaaggccgtg
      481 caccgcgcgc tctggcgttt ctgcagtgcg tcgaaaacga agtggcagct tatatcacca
      541 tgaaagtggt tatggatatg ctgaataccg acgcgacgct gcaggccatt gcaatgtccg
      601 tcgccgaacg tatcgaagat caagtgcgct tttcaaaact ggaaggtcat gcggccaaat
      661 acttcgaaaa agtgaaaaaa tcactgaaag cgtcgcgtac caaaagctac cgccatgccc
      721 acaacgttgc tgtcgtggcg gaaaaatccg tcgccgaaaa agatgcagac tttgatcgct
      781 gggaagcgtg gccgaaagaa acgcagctgc aaattggcac cacgctgctg gaaatcctgg
      841 aaggcagcgt gttttataat ggcgagccgg tgttcatgcg tgccatgcgc acctacggtg
      901 gcaaaacgat ttattacctg cagaccagcg aaagcgtggg ccaatggatc tctgccttca
      961 aagaacacgt cgcacagctg agtccggctt atgcgccgtg tgtgattccg ccgcgtccgt
     1021 ggcgtacccc gtttaacggt ggcttccata cggaaaaagt ggcgagccgt atccgcctgg
     1081 ttaaaggcaa tcgtgaacac gtccgcaaac tgacccagaa acaaatgccg aaagtgtata
     1141 aagccattaa cgcactgcag aatacgcagt ggcaaatcaa caaagatgtt ctggcggtca
     1201 ttgaagaagt gatccgtctg gacctgggtt acggcgtgcc gagctttaaa ccgctgatcg
     1261 ataaagaaaa caaaccggcc aatccggttc cggtcgaatt tcagcatctg cgtggtcgcg
     1321 aactgaaaga aatgctgagt ccggaacaat ggcagcaatt tattaactgg aaaggcgaat
     1381 gcgcccgtct gtataccgca gaaacgaaac gcggtagtaa atccgcagct gttgtccgta
     1441 tggttggcca ggctcgcaaa tactctgcgt tcgaaagtat ctacttcgtg tacgcgatgg
     1501 attcacgttc gcgcgtgtac gttcagagca gcaccctgag cccgcaatct aacgacctgg
     1561 gtaaagccct gctgcgtttt acggaaggtc gcccggtgaa tggcgttgaa gcactgaaat
     1621 ggttctgtat taacggtgct aatctgtggg gctgggataa gaaaaccttt gatgtgcgtg
     1681 tgtctaacgt tctggatgaa gaatttcagg acatgtgccg tgatatcgca gccgacccgc
     1741 tgacctttac gcaatgggcc aaagcagatg ctccgtatga atttctggct tggtgtttcg
     1801 aatatgcgca gtacctggac ctggttgatg aaggccgtgc cgatgaattt cgcacccatc
     1861 tgccggtgca ccaggacggt agctgctctg gcattcaaca ttactctgca atgctgcgtg
     1921 atgaagttgg tgcgaaagcc gtcaacctga aaccgagtga tgcaccgcag gacatttatg
     1981 gcgcagttgc tcaagtggtt attaagaaaa acgcgctgta catggatgct gatgacgcga
     2041 ccacgtttac cagtggttcc gtgaccctgt ccggcacgga actgcgtgcg atggcctccg
     2101 catgggatag cattggcatc acccgctcac tgaccaaaaa accagtgatg acgctgccgt
     2161 atggctcgac ccgtctgacg tgtcgcgaaa gcgtgattga ttacatcgtt gacctggaag
     2221 aaaaagaagc acagaaagct gtcgcggaag gtcgtaccgc taacaaagtg cacccgtttg
     2281 aagatgaccg ccaggattat ctgaccccgg gcgcagctta taattacatg acggcgctga
     2341 tttggccgtc aatctcggaa gtcgtgaaag ctccgattgt tgcgatgaaa atgatccgtc
     2401 agctggcccg ctttgcggcc aaacgcaacg aaggtctgat gtacaccctg ccgacgggct
     2461 tcattctgga acagaaaatc atggcgaccg aaatgctgcg tgtgcgcacg tgcctgatgg
     2521 gtgatattaa aatgtctctg caggtggaaa ccgacatcgt ggatgaagca gctatgatgg
     2581 gtgcggcggc cccgaatttt gttcatggcc acgatgcgtc gcatctgatt ctgaccgttt
     2641 gtgaactggt ggataaaggt gtgacgagca ttgcagtgat ccatgattcc ttcggcaccc
     2701 acgcggacaa caccctgacg ctgcgtgtgg ccctgaaagg ccagatggtt gcaatgtata
     2761 ttgatggcaa tgctctgcaa aaactgctgg aagaacacga agaacgctgg atggtggaca
     2821 ccggcatcga agtgccggaa caaggcgaat ttgacctgaa tgaaatcatg gacagtgaat
     2881 acgtctttgc ctaataagga tccggcgccg ctcattcgct aatcgccacg acgcgtagtc
     2941 gacccgccaa ttcccatggg gcgcgcctgc tgccaccgct gagcaataac tagcataacc
     3001 ccttggggcc tctaaacggg tcttgagggg ttttttgcca ggcatcaaat aaaacgaaag
     3061 gctcagtcgg aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     3121 agtaggacaa atccgccggg agcggatttg aacgttgtga agcaacggcc cggagggtgg
     3181 cgggcaggac gcccgccata aactgccagg catcaaacta agcagaaggc catcctgacg
     3241 gatggccttt ttgcgtttca gatctaccgg taaaccagca atagacataa gcggctattt
     3301 aacgaccctg ccctgaaccg acgacaagct gacgaccggg tctccgcaag tggcactttt
     3361 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3421 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     3481 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     3541 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     3601 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     3661 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     3721 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     3781 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cggtcgctgt taaaaggaca
     3841 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     3901 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     3961 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     4021 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     4081 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     4141 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     4201 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcatactctt
     4261 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4321 tgaatgtatt tagaaaaata aacaaatagg catgctagcg cagaaacgtc ctagaagatg
     4381 ccaggaggat acttagcaga gagacaataa ggccggagcg aagccgtttt tccataggct
     4441 ccgcccccct gacgaacatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     4501 aggactataa agataccagg cgtttccccc tgatggctcc ctcttgcgct ctcctgttcc
     4561 cgtcctgcgg cgtccgtgtt gtggtggagg ctttacccaa atcaccacgt cccgttccgt
     4621 gtagacagtt cgctccaagc tgggctgtgt gcaagaaccc cccgttcagc ccgactgctg
     4681 cgccttatcc ggtaactatc atcttgagtc caacccggaa agacacgaca aaacgccact
     4741 ggcagcagcc attggtaact gagaattagt ggatttagat atcgagagtc ttgaagtggt
     4801 ggcctaacag aggctacact gaaaggacag tatttggtat ctgcgctcca ctaaagccag
     4861 ttaccaggtt aagcagttcc ccaactgact taaccttcga tcaaaccgcc tccccaggcg
     4921 gttttttcgt ttacagagca ggagattacg acgatcgtaa aaggatctca agaagatcct
     4981 ttacggattc ccgacaccat cactctagat ttcagtgcaa tttatctctt caaatgtagc
     5041 acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc
     5101 caccggaagg agctgactgg gttgctccta ggtctgattc gttaccaatt atgacaactt
     5161 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
     5221 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
     5281 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
     5341 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
     5401 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
     5461 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
     5521 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
     5581 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
     5641 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
     5701 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
     5761 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
     5821 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
     5881 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
     5941 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
     6001 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gaggaagaaa
     6061 ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc
     6121 taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca
     6181 tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt
     6241 tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta gcggttccta
     6301 cctgacgctt tttatcgcaa ctctctactg tttctccata cc
//
