LOCUS       40924_39293        4483 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA227D, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4483)
  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  JOURNAL   Unpublished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 4483)
  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 4483)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4483)
  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            "Unpublished"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..4483
                     /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..801
                     /codon_start=1
                     /label=DsRed2
                     /note="improved tetrameric variant of DsRed fluorescent
                     protein"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     terminator      815..909
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1041..1853
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2023..2131
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2195..3340)
                     /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(3733..4264)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(4391..4477)
                     /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 cctcctccga gaacgtcatc accgagttca tgcgcttcaa ggtgcgcatg
      181 gagggcaccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
      241 gagggccaca acaccgtgaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
      301 gacatcctgt ccccccagtt ccagtacggc tccaaggtgt acgtgaagca ccccgccgac
      361 atccccgact acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
      421 ttcgaggacg gcggcgtggc gaccgtgacc caggactcct ccctgcaaga cggctgcttc
      481 atctacaagg tgaagttcat cggcgtgaac ttcccctccg acggccccgt gatgcagaag
      541 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag
      601 ggcgagaccc acaaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag
      661 tccatctaca tggccaagaa gcccgtgcag ctgcccggct actactacgt ggacgccaag
      721 ctggacatca cctcccacaa cgaggactac accatcgtgg agcagtacga gcgcaccgag
      781 ggccgccacc acctgttcct gtagactagt cttggactcc tgttgataga tccagtaatg
      841 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
      901 ggtgagaatc caggggtccc caataattac gatttaaatt tgtgtctcaa aatctctgat
      961 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     1021 acagtaatac aaggggtgtt atgagccata ttcagcgtga aacgagctgt agccgtccgc
     1081 gtctgaacag caacatggat gcggatctgt atggctataa atgggcgcgt gataacgtgg
     1141 gtcagagcgg cgcgaccatt tatcgtctgt atggcaaacc ggatgcgccg gaactgtttc
     1201 tgaaacatgg caaaggcagc gtggcgaacg atgtgaccga tgaaatggtg cgtctgaact
     1261 ggctgaccga atttatgccg ctgccgacca ttaaacattt tattcgcacc ccggatgatg
     1321 cgtggctgct gaccaccgcg attccgggca aaaccgcgtt tcaggtgctg gaagaatatc
     1381 cggatagcgg cgaaaacatt gtggatgcgc tggccgtgtt tctgcgtcgt ctgcatagca
     1441 ttccggtgtg caactgcccg tttaacagcg atcgtgtgtt tcgtctggcc caggcgcaga
     1501 gccgtatgaa caacggcctg gtggatgcga gcgattttga tgatgaacgt aacggctggc
     1561 cggtggaaca ggtgtggaaa gaaatgcata aactgctgcc gtttagcccg gatagcgtgg
     1621 tgacccacgg cgattttagc ctggataacc tgattttcga tgaaggcaaa ctgattggct
     1681 gcattgatgt gggccgtgtg ggcattgcgg atcgttatca ggatctggcc attctgtgga
     1741 actgcctggg cgaatttagc ccgagcctgc aaaaacgtct gtttcagaaa tatggcattg
     1801 ataatccgga tatgaacaaa ctgcaatttc atctgatgct ggatgaattt ttctaataat
     1861 taattggacc gcggtccgcg cgttgtcctt ttccgctgca taaccctgct tcggggtcat
     1921 tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag
     1981 ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt
     2041 aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct
     2101 caacgggaat cctgctctgc gaggctggcc gtaggccggc cgcgatgcag gtggctgctg
     2161 aacccccagc cggaactgac cccacaaggc cctagcgttt gcaatgcacc aggtcatcat
     2221 tgacccaggc gtgttccacc aggccgctgc ctcgcaactc ttcgcaggct tcgccgacct
     2281 gctcgcgcca cttcttcacg cgggtggaat ccgatccgca catgaggcgg aaggtttcca
     2341 gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc gaacatccgt cgggccgtcg
     2401 gcgacagctt gcggtacttc tcccatatga atttcgtgta gtggtcgcca gcaaacagca
     2461 cgacgatttc ctcgtcgatc aggacctggc aacgggacgt tttcttgcca cggtccagga
     2521 cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc aacgcggtcg gacgtgaagc
     2581 ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc cttcgtgtaa taccggccat
     2641 tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt gaaggtgatc ggctcgccga
     2701 taggggtgcg cttcgcgtac tccaacacct gctgccacac cagttcgtca tcgtcggccc
     2761 gcagctcgac gccggtgtag gtgatcttca cgtccttgtt gacgtggaaa atgaccttgt
     2821 tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt gaacagggca gagcgggccg
     2881 tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc aatatcgaac aaggaaagct
     2941 gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc ggcctgcttg gcttcgctga
     3001 cctgttttgc caggtcctcg ccggcggttt ttcgcttctt ggtcgtcata gttcctcgcg
     3061 tgtcgatggt catcgacttc gccaaacctg ccgcctcctg ttcgagacga cgcgaacgct
     3121 ccacggcggc cgatggcgcg ggcagggcag ggggagccag ttgcacgctg tcgcgctcga
     3181 tcttggccgt agcttgctgg actatcgagc cgacggactg gaaggtttcg cggggcgcac
     3241 gcatgacggt gcggcttgcg atggtttcgg catcctcggc ggaaaacccc gcgtcgatca
     3301 gttcttgcct gtatgccttc cggtcaaacg tccgattcat tcaccctcct tgcgggattg
     3361 ccccggaatt aattccccgg atcgatccgt cgatcttgat cccctgcgcc atcagatcct
     3421 tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc
     3481 cccagctggc aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca
     3541 tgtaagccca ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat
     3601 agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt
     3661 ggctgccatt tttggggtga ggccgttcgc ggccgagggg cgcagcccct ggggggatgg
     3721 gaggcccgcg ttagcgggcc gggagggttc gagaaggggg ggcacccccc ttcggcgtgc
     3781 gcggtcacgc gcacagggcg cagccctggt taaaaacaag gtttataaat attggtttaa
     3841 aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc
     3901 tggattttct gcctgtggac agcccctcaa atgtcaatag gtgcgcccct catctgtcag
     3961 cactctgccc ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc gcgcccctca
     4021 agtgtcaata ccgcagggca cttatcccca ggcttgtcca catcatctgt gggaaactcg
     4081 cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca gctccacgtc gccggccgaa
     4141 atcgagcctg cccctcatct gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa
     4201 cgtccgcccc tcatctgtca gtgagggcca agttttccgc gaggtatcca caacgccggc
     4261 ggccctacat ggctctgctg tagtgagtgg gttgcgctcc ggcagcggtc ctgatccccc
     4321 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggcgcgc ccagctgtct
     4381 agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     4441 aggcccagtc tttcgactga gcctttcgtt ttatttgatg cct
//
