LOCUS       40924_39263        3823 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA221, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3823)
  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  JOURNAL   Unpublished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 3823)
  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 3823)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3823)
  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            "Unpublished"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..3823
                     /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"
     primer_bind     complement(121..144)
                     /label=F24
                     /note="F24"
     terminator      155..249
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             381..1193
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            1363..1471
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(1535..2680)
                     /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(3073..3604)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(3731..3817)
                     /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 ttgcggccgc
      121 gtcgtgactg ggaaaaccct ggcgactagt cttggactcc tgttgataga tccagtaatg
      181 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
      241 ggtgagaatc caggggtccc caataattac gatttaaatt tgtgtctcaa aatctctgat
      301 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
      361 acagtaatac aaggggtgtt atgagccata ttcagcgtga aacgagctgt agccgtccgc
      421 gtctgaacag caacatggat gcggatctgt atggctataa atgggcgcgt gataacgtgg
      481 gtcagagcgg cgcgaccatt tatcgtctgt atggcaaacc ggatgcgccg gaactgtttc
      541 tgaaacatgg caaaggcagc gtggcgaacg atgtgaccga tgaaatggtg cgtctgaact
      601 ggctgaccga atttatgccg ctgccgacca ttaaacattt tattcgcacc ccggatgatg
      661 cgtggctgct gaccaccgcg attccgggca aaaccgcgtt tcaggtgctg gaagaatatc
      721 cggatagcgg cgaaaacatt gtggatgcgc tggccgtgtt tctgcgtcgt ctgcatagca
      781 ttccggtgtg caactgcccg tttaacagcg atcgtgtgtt tcgtctggcc caggcgcaga
      841 gccgtatgaa caacggcctg gtggatgcga gcgattttga tgatgaacgt aacggctggc
      901 cggtggaaca ggtgtggaaa gaaatgcata aactgctgcc gtttagcccg gatagcgtgg
      961 tgacccacgg cgattttagc ctggataacc tgattttcga tgaaggcaaa ctgattggct
     1021 gcattgatgt gggccgtgtg ggcattgcgg atcgttatca ggatctggcc attctgtgga
     1081 actgcctggg cgaatttagc ccgagcctgc aaaaacgtct gtttcagaaa tatggcattg
     1141 ataatccgga tatgaacaaa ctgcaatttc atctgatgct ggatgaattt ttctaataat
     1201 taattggacc gcggtccgcg cgttgtcctt ttccgctgca taaccctgct tcggggtcat
     1261 tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag
     1321 ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt
     1381 aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct
     1441 caacgggaat cctgctctgc gaggctggcc gtaggccggc cgcgatgcag gtggctgctg
     1501 aacccccagc cggaactgac cccacaaggc cctagcgttt gcaatgcacc aggtcatcat
     1561 tgacccaggc gtgttccacc aggccgctgc ctcgcaactc ttcgcaggct tcgccgacct
     1621 gctcgcgcca cttcttcacg cgggtggaat ccgatccgca catgaggcgg aaggtttcca
     1681 gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc gaacatccgt cgggccgtcg
     1741 gcgacagctt gcggtacttc tcccatatga atttcgtgta gtggtcgcca gcaaacagca
     1801 cgacgatttc ctcgtcgatc aggacctggc aacgggacgt tttcttgcca cggtccagga
     1861 cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc aacgcggtcg gacgtgaagc
     1921 ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc cttcgtgtaa taccggccat
     1981 tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt gaaggtgatc ggctcgccga
     2041 taggggtgcg cttcgcgtac tccaacacct gctgccacac cagttcgtca tcgtcggccc
     2101 gcagctcgac gccggtgtag gtgatcttca cgtccttgtt gacgtggaaa atgaccttgt
     2161 tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt gaacagggca gagcgggccg
     2221 tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc aatatcgaac aaggaaagct
     2281 gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc ggcctgcttg gcttcgctga
     2341 cctgttttgc caggtcctcg ccggcggttt ttcgcttctt ggtcgtcata gttcctcgcg
     2401 tgtcgatggt catcgacttc gccaaacctg ccgcctcctg ttcgagacga cgcgaacgct
     2461 ccacggcggc cgatggcgcg ggcagggcag ggggagccag ttgcacgctg tcgcgctcga
     2521 tcttggccgt agcttgctgg actatcgagc cgacggactg gaaggtttcg cggggcgcac
     2581 gcatgacggt gcggcttgcg atggtttcgg catcctcggc ggaaaacccc gcgtcgatca
     2641 gttcttgcct gtatgccttc cggtcaaacg tccgattcat tcaccctcct tgcgggattg
     2701 ccccggaatt aattccccgg atcgatccgt cgatcttgat cccctgcgcc atcagatcct
     2761 tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc
     2821 cccagctggc aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca
     2881 tgtaagccca ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat
     2941 agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt
     3001 ggctgccatt tttggggtga ggccgttcgc ggccgagggg cgcagcccct ggggggatgg
     3061 gaggcccgcg ttagcgggcc gggagggttc gagaaggggg ggcacccccc ttcggcgtgc
     3121 gcggtcacgc gcacagggcg cagccctggt taaaaacaag gtttataaat attggtttaa
     3181 aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc
     3241 tggattttct gcctgtggac agcccctcaa atgtcaatag gtgcgcccct catctgtcag
     3301 cactctgccc ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc gcgcccctca
     3361 agtgtcaata ccgcagggca cttatcccca ggcttgtcca catcatctgt gggaaactcg
     3421 cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca gctccacgtc gccggccgaa
     3481 atcgagcctg cccctcatct gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa
     3541 cgtccgcccc tcatctgtca gtgagggcca agttttccgc gaggtatcca caacgccggc
     3601 ggccctacat ggctctgctg tagtgagtgg gttgcgctcc ggcagcggtc ctgatccccc
     3661 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggcgcgc ccagctgtct
     3721 agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     3781 aggcccagtc tttcgactga gcctttcgtt ttatttgatg cct
//
