LOCUS       40924_39298        4516 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA227R, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4516)
  AUTHORS   Silva-Rocha R, Martinez-Garcia E, Chavarria M, Calles B, 
            Arce-Rodriguez A, de las Heras A, Paez-Espino D, Durante-Rodriguez 
            G, Kim J, Nikel PI, Platero R, de Lorenzo V.
  TITLE     The Standard European Vector Architecture (SEVA): a coherent 
            platform for analysis and deployment of complex prokaryotic 
            phenotypes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4516)
  AUTHORS   Silva-Rocha R, Martinez-Garcia E, Chavarria M, Calles B, 
            Arce-Rodriguez A, de las Heras A, Paez-Espino D, Durante-Rodriguez 
            G, Kim J, Nikel PI, Platero R, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-AUG-2012) Systems Biology Program, Centro Nacional de 
            Biotecnologia-CSIC, C/ Darwin 3, Campus de Cantoblanco, Madrid, 
            Madrid 28049, Spain
REFERENCE   3  (bases 1 to 4516)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4516)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-AUG-2012) Systems Biology Program, Centro Nacional de 
            Biotecnologia-CSIC, C/ Darwin 3, Campus de Cantoblanco, Madrid, 
            Madrid 28049, Spain"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: ApE v. v2.0.40
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..32
                     /label=R24
                     /note="R24"
     misc_feature    56..112
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             127..834
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     terminator      848..942
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1074..1886
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2056..2164
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2228..3373)
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(3766..4297)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(4424..4510)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc ttaggaggaa
      121 aaacatatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
      181 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      241 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
      301 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
      361 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
      421 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
      481 caagacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
      541 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
      601 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
      661 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
      721 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
      781 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaact
      841 agtcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
      901 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaggggt ccccaataat
      961 tacgatttaa atttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     1021 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     1081 atattcagcg tgaaacgagc tgtagccgtc cgcgtctgaa cagcaacatg gatgcggatc
     1141 tgtatggcta taaatgggcg cgtgataacg tgggtcagag cggcgcgacc atttatcgtc
     1201 tgtatggcaa accggatgcg ccggaactgt ttctgaaaca tggcaaaggc agcgtggcga
     1261 acgatgtgac cgatgaaatg gtgcgtctga actggctgac cgaatttatg ccgctgccga
     1321 ccattaaaca ttttattcgc accccggatg atgcgtggct gctgaccacc gcgattccgg
     1381 gcaaaaccgc gtttcaggtg ctggaagaat atccggatag cggcgaaaac attgtggatg
     1441 cgctggccgt gtttctgcgt cgtctgcata gcattccggt gtgcaactgc ccgtttaaca
     1501 gcgatcgtgt gtttcgtctg gcccaggcgc agagccgtat gaacaacggc ctggtggatg
     1561 cgagcgattt tgatgatgaa cgtaacggct ggccggtgga acaggtgtgg aaagaaatgc
     1621 ataaactgct gccgtttagc ccggatagcg tggtgaccca cggcgatttt agcctggata
     1681 acctgatttt cgatgaaggc aaactgattg gctgcattga tgtgggccgt gtgggcattg
     1741 cggatcgtta tcaggatctg gccattctgt ggaactgcct gggcgaattt agcccgagcc
     1801 tgcaaaaacg tctgtttcag aaatatggca ttgataatcc ggatatgaac aaactgcaat
     1861 ttcatctgat gctggatgaa tttttctaat aattaattgg accgcggtcc gcgcgttgtc
     1921 cttttccgct gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc
     1981 ctttttcgca cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat
     2041 ccaacggcgt cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt
     2101 cttcactgtc ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg
     2161 gccgtaggcc ggccgcgatg caggtggctg ctgaaccccc agccggaact gaccccacaa
     2221 ggccctagcg tttgcaatgc accaggtcat cattgaccca ggcgtgttcc accaggccgc
     2281 tgcctcgcaa ctcttcgcag gcttcgccga cctgctcgcg ccacttcttc acgcgggtgg
     2341 aatccgatcc gcacatgagg cggaaggttt ccagcttgag cgggtacggc tcccggtgcg
     2401 agctgaaata gtcgaacatc cgtcgggccg tcggcgacag cttgcggtac ttctcccata
     2461 tgaatttcgt gtagtggtcg ccagcaaaca gcacgacgat ttcctcgtcg atcaggacct
     2521 ggcaacggga cgttttcttg ccacggtcca ggacgcggaa gcggtgcagc agcgacaccg
     2581 attccaggtg cccaacgcgg tcggacgtga agcccatcgc cgtcgcctgt aggcgcgaca
     2641 ggcattcctc ggccttcgtg taataccggc cattgatcga ccagcccagg tcctggcaaa
     2701 gctcgtagaa cgtgaaggtg atcggctcgc cgataggggt gcgcttcgcg tactccaaca
     2761 cctgctgcca caccagttcg tcatcgtcgg cccgcagctc gacgccggtg taggtgatct
     2821 tcacgtcctt gttgacgtgg aaaatgacct tgttttgcag cgcctcgcgc gggattttct
     2881 tgttgcgcgt ggtgaacagg gcagagcggg ccgtgtcgtt tggcatcgct cgcatcgtgt
     2941 ccggccacgg cgcaatatcg aacaaggaaa gctgcatttc cttgatctgc tgcttcgtgt
     3001 gtttcagcaa cgcggcctgc ttggcttcgc tgacctgttt tgccaggtcc tcgccggcgg
     3061 tttttcgctt cttggtcgtc atagttcctc gcgtgtcgat ggtcatcgac ttcgccaaac
     3121 ctgccgcctc ctgttcgaga cgacgcgaac gctccacggc ggccgatggc gcgggcaggg
     3181 cagggggagc cagttgcacg ctgtcgcgct cgatcttggc cgtagcttgc tggactatcg
     3241 agccgacgga ctggaaggtt tcgcggggcg cacgcatgac ggtgcggctt gcgatggttt
     3301 cggcatcctc ggcggaaaac cccgcgtcga tcagttcttg cctgtatgcc ttccggtcaa
     3361 acgtccgatt cattcaccct ccttgcggga ttgccccgga attaattccc cggatcgatc
     3421 cgtcgatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
     3481 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg gttcgcttgc
     3541 tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt
     3601 tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg
     3661 tcagcaccgt ttctgcggac tggctttcta cgtggctgcc atttttgggg tgaggccgtt
     3721 cgcggccgag gggcgcagcc cctgggggga tgggaggccc gcgttagcgg gccgggaggg
     3781 ttcgagaagg gggggcaccc cccttcggcg tgcgcggtca cgcgcacagg gcgcagccct
     3841 ggttaaaaac aaggtttata aatattggtt taaaagcagg ttaaaagaca ggttagcggt
     3901 ggccgaaaaa cgggcggaaa cccttgcaaa tgctggattt tctgcctgtg gacagcccct
     3961 caaatgtcaa taggtgcgcc cctcatctgt cagcactctg cccctcaagt gtcaaggatc
     4021 gcgcccctca tctgtcagta gtcgcgcccc tcaagtgtca ataccgcagg gcacttatcc
     4081 ccaggcttgt ccacatcatc tgtgggaaac tcgcgtaaaa tcaggcgttt tcgccgattt
     4141 gcgaggctgg ccagctccac gtcgccggcc gaaatcgagc ctgcccctca tctgtcaacg
     4201 ccgcgccggg tgagtcggcc cctcaagtgt caacgtccgc ccctcatctg tcagtgaggg
     4261 ccaagttttc cgcgaggtat ccacaacgcc ggcggcccta catggctctg ctgtagtgag
     4321 tgggttgcgc tccggcagcg gtcctgatcc cccgcagaaa aaaaggatct caagaagatc
     4381 ctttgatctt ttctacggcg cgcccagctg tctagggcgg cggatttgtc ctactcagga
     4441 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
     4501 gttttatttg atgcct
//
