LOCUS       40924_39273        6964 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA225, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6964)
  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  JOURNAL   Unpublished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 6964)
  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 6964)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6964)
  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            "Unpublished"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..6964
                     /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..3198
                     /codon_start=1
                     /label=lacZ
                     /note="beta-galactosidase"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     terminator      3296..3390
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3522..4334
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            4504..4612
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(4676..5821)
                     /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(6214..6745)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(6872..6958)
                     /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 aaacatatga ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa
      181 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
      241 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
      301 tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat
      361 cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg
      421 cccatctaca ccaacgtaac ctatcccatt acggtcaatc cgccgtttgt tcccacggag
      481 aatccgacgg gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc
      541 cagacgcgaa ttatttttga tggcgttaac tcggcgtttc atctgtggtg caacgggcgc
      601 tgggtcggtt acggccagga cagtcgtttg ccgtctgaat ttgacctgag cgcattttta
      661 cgcgccggag aaaaccgcct cgcggtgatg gtgctgcgtt ggagtgacgg cagttatctg
      721 gaagatcagg atatgtggcg gatgagcggc attttccgtg acgtgtcgtt gctgcataaa
      781 ccgactacac aaatcagcga tttccatgtt gccactcgct ttaatgatga tttcagccgc
      841 gctgtactgg aggctgaagt tcagatgtgc ggcgagttgc gtgactacct acgggtaaca
      901 gtttctttat ggcagggtga aacgcaggtc gccagcggca ccgcgccttt cggcggtgaa
      961 attatcgatg agcgtggtgg ttatgccgat cgcgtcacac tacgtctgaa cgtcgaaaac
     1021 ccgaaactgt ggagcgccga aatcccgaat ctctatcgtg cggtggttga actgcacacc
     1081 gccgacggca cgctgattga agcagaagcc tgcgatgtcg gtttccgcga ggtgcggatt
     1141 gaaaatggtc tgctgctgct gaacggcaag ccgttgctga ttcgaggcgt taaccgtcac
     1201 gagcatcatc ctctgcatgg tcaggtcatg gatgagcaga cgatggtgca ggatatcctg
     1261 ctgatgaagc agaacaactt taacgccgtg cgctgttcgc attatccgaa ccatccgctg
     1321 tggtacacgc tgtgcgaccg ctacggcctg tatgtggtgg atgaagccaa tattgaaacc
     1381 cacggcatgg tgccaatgaa tcgtctgacc gatgatccgc gctggctacc ggcgatgagc
     1441 gaacgcgtaa cgcgaatggt gcagcgcgat cgtaatcacc cgagtgtgat catctggtcg
     1501 ctggggaatg aatcaggcca cggcgctaat cacgacgcgc tgtatcgctg gatcaaatct
     1561 gtcgatcctt cccgcccggt gcagtatgaa ggcggcggag ccgacaccac ggccaccgat
     1621 attatttgcc cgatgtacgc gcgcgtggat gaagaccagc ccttcccggc tgtgccgaaa
     1681 tggtccatca aaaaatggct ttcgctacct ggagagacgc gcccgctgat cctttgcgaa
     1741 tacgcccacg cgatgggtaa cagtcttggc ggtttcgcta aatactggca ggcgtttcgt
     1801 cagtatcccc gtttacaggg cggcttcgtc tgggactggg tggatcagtc gctgattaaa
     1861 tatgatgaaa acggcaaccc gtggtcggct tacggcggtg attttggcga tacgccgaac
     1921 gatcgccagt tctgtatgaa cggtctggtc tttgccgacc gcacgccgca tccagcgctg
     1981 acggaagcaa aacaccagca gcagtttttc cagttccgtt tatccgggca aaccatcgaa
     2041 gtgaccagcg aatacctgtt ccgtcatagc gataacgagc tgctgcactg gatggtggcg
     2101 ctggatggta agccgctggc aagcggtgaa gtgcctctgg atgtcgctcc acaaggtaaa
     2161 cagttgattg aactgcctga actaccgcag ccggagagcg ccgggcaact ctggctcaca
     2221 gtacgcgtag tgcaaccgaa cgcgaccgca tggtcagaag ccgggcacat cagcgcctgg
     2281 cagcagtggc gtctggcgga aaacctcagt gtgacgctcc ccgccgcgtc ccacgccatc
     2341 ccgcatctga ccaccagcga aatggatttt tgcatcgagc tgggtaataa gcgttggcaa
     2401 tttaaccgcc agtcaggctt tctttcacag atgtggattg gcgataaaaa acaactgctg
     2461 acgccgctgc gcgatcagtt cacccgtgca ccgctggata acgacattgg cgtaagtgaa
     2521 gcgacccgca ttgaccctaa cgcctgggtc gaacgctgga aggcggcggg ccattaccag
     2581 gccgaagcag cgttgttgca gtgcacggca gatacacttg ctgatgcggt gctgattacg
     2641 accgctcacg cgtggcagca tcaggggaaa accttattta tcagccggaa aacctaccgg
     2701 attgatggta gtggtcaaat ggcgattacc gttgatgttg aagtggcgag cgatacaccg
     2761 catccggcgc ggattggcct gaactgccag ctggcgcagg tagcagagcg ggtaaactgg
     2821 ctcggattag ggccgcaaga aaactatccc gaccgcctta ctgccgcctg ttttgaccgc
     2881 tgggatctgc cattgtcaga catgtatacc ccgtacgtct tcccgagcga aaacggtctg
     2941 cgctgcggga cgcgcgaatt gaattatggc ccacaccagt ggcgcggcga cttccagttc
     3001 aacatcagcc gctacagtca acagcaactg atggaaacca gccatcgcca tctgctgcac
     3061 gcggaagaag gcacatggct gaatatcgac ggtttccata tggggattgg tggcgacgac
     3121 tcctggagcc cgtcagtatc ggcggagttc cagctgagcg ccggtcgcta ccattaccag
     3181 ttggtctggt gtcaaaaata ataataaccg ggcaggccat gtctgcccgt atttcgcgta
     3241 aggaaatcca ttatgtacta tttaaaatca agctccttgc aggccactag tcttggactc
     3301 ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt
     3361 tgccgccggg cgttttttat tggtgagaat ccaggggtcc ccaataatta cgatttaaat
     3421 ttgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     3481 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcagcgtg
     3541 aaacgagctg tagccgtccg cgtctgaaca gcaacatgga tgcggatctg tatggctata
     3601 aatgggcgcg tgataacgtg ggtcagagcg gcgcgaccat ttatcgtctg tatggcaaac
     3661 cggatgcgcc ggaactgttt ctgaaacatg gcaaaggcag cgtggcgaac gatgtgaccg
     3721 atgaaatggt gcgtctgaac tggctgaccg aatttatgcc gctgccgacc attaaacatt
     3781 ttattcgcac cccggatgat gcgtggctgc tgaccaccgc gattccgggc aaaaccgcgt
     3841 ttcaggtgct ggaagaatat ccggatagcg gcgaaaacat tgtggatgcg ctggccgtgt
     3901 ttctgcgtcg tctgcatagc attccggtgt gcaactgccc gtttaacagc gatcgtgtgt
     3961 ttcgtctggc ccaggcgcag agccgtatga acaacggcct ggtggatgcg agcgattttg
     4021 atgatgaacg taacggctgg ccggtggaac aggtgtggaa agaaatgcat aaactgctgc
     4081 cgtttagccc ggatagcgtg gtgacccacg gcgattttag cctggataac ctgattttcg
     4141 atgaaggcaa actgattggc tgcattgatg tgggccgtgt gggcattgcg gatcgttatc
     4201 aggatctggc cattctgtgg aactgcctgg gcgaatttag cccgagcctg caaaaacgtc
     4261 tgtttcagaa atatggcatt gataatccgg atatgaacaa actgcaattt catctgatgc
     4321 tggatgaatt tttctaataa ttaattggac cgcggtccgc gcgttgtcct tttccgctgc
     4381 ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg
     4441 atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca
     4501 gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc
     4561 ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cgtaggccgg
     4621 ccgcgatgca ggtggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt
     4681 tgcaatgcac caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact
     4741 cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc
     4801 acatgaggcg gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt
     4861 cgaacatccg tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt
     4921 agtggtcgcc agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg
     4981 ttttcttgcc acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc
     5041 caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg
     5101 ccttcgtgta ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg
     5161 tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca
     5221 ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt
     5281 tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg
     5341 tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg
     5401 caatatcgaa caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg
     5461 cggcctgctt ggcttcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct
     5521 tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct
     5581 gttcgagacg acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca
     5641 gttgcacgct gtcgcgctcg atcttggccg tagcttgctg gactatcgag ccgacggact
     5701 ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg
     5761 cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca
     5821 ttcaccctcc ttgcgggatt gccccggaat taattccccg gatcgatccg tcgatcttga
     5881 tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt
     5941 cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg tccataaaac
     6001 cgcccagtct agctatcgcc atgtaagccc actgcaagct acctgctttc tctttgcgct
     6061 tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt
     6121 ctgcggactg gctttctacg tggctgccat ttttggggtg aggccgttcg cggccgaggg
     6181 gcgcagcccc tggggggatg ggaggcccgc gttagcgggc cgggagggtt cgagaagggg
     6241 gggcaccccc cttcggcgtg cgcggtcacg cgcacagggc gcagccctgg ttaaaaacaa
     6301 ggtttataaa tattggttta aaagcaggtt aaaagacagg ttagcggtgg ccgaaaaacg
     6361 ggcggaaacc cttgcaaatg ctggattttc tgcctgtgga cagcccctca aatgtcaata
     6421 ggtgcgcccc tcatctgtca gcactctgcc cctcaagtgt caaggatcgc gcccctcatc
     6481 tgtcagtagt cgcgcccctc aagtgtcaat accgcagggc acttatcccc aggcttgtcc
     6541 acatcatctg tgggaaactc gcgtaaaatc aggcgttttc gccgatttgc gaggctggcc
     6601 agctccacgt cgccggccga aatcgagcct gcccctcatc tgtcaacgcc gcgccgggtg
     6661 agtcggcccc tcaagtgtca acgtccgccc ctcatctgtc agtgagggcc aagttttccg
     6721 cgaggtatcc acaacgccgg cggccctaca tggctctgct gtagtgagtg ggttgcgctc
     6781 cggcagcggt cctgatcccc cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6841 ctacggcgcg cccagctgtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg
     6901 acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat
     6961 gcct
//
