LOCUS       40924_39278        6939 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA225T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6939)
  AUTHORS   Silva-Rocha R, Martinez-Garcia E, Calles B, Chavarria M, 
            Arce-Rodriguez A, de Las Heras A, Paez-Espino AD, Durante-Rodriguez 
            G, Kim J, Nikel PI, Platero R, de Lorenzo V.
  TITLE     The Standard European Vector Architecture (SEVA): a coherent 
            platform for the analysis and deployment of complex prokaryotic 
            phenotypes
  JOURNAL   Nucleic Acids Res. 41 (DATABASE ISSUE), D666-D675 (2013)
  PUBMED    23180763
REFERENCE   2  (bases 1 to 6939)
  AUTHORS   Silva-Rocha R, Martinez-Garcia E, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-APR-2013) Systems Biology Program, CNB-CSIC, Darwin 3,
            Madrid, Madrid 28029, Spain
REFERENCE   3  (bases 1 to 6939)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6939)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 41 (DATABASE ISSUE), D666-D675 (2013)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-APR-2013) Systems Biology Program, CNB-CSIC, Darwin 3, Madrid, 
            Madrid 28029, 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..6939
                     /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             111..3173
                     /codon_start=1
                     /label=lacZ
                     /note="beta-galactosidase"
                     /translation="LTDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRP
                     SQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYP
                     ITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDS
                     RLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISD
                     FHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERG
                     GYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLL
                     LLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTL
                     CDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGN
                     ESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWS
                     IKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKY
                     DENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIE
                     VTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWL
                     TVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKR
                     WQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAG
                     HYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVA
                     SDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFP
                     SENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHM
                     GIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     terminator      3271..3365
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3497..4309
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            4479..4587
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(4651..5796)
                     /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(6189..6720)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(6847..6933)
                     /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 ttgacggatt
      121 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
      181 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
      241 gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg tttccggcac
      301 cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat actgtcgtcg
      361 tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac gtaacctatc
      421 ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt tactcgctca
      481 catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt tttgatggcg
      541 ttaactcggc gtttcatctg tggtgcaacg ggcgctgggt cggttacggc caggacagtc
      601 gtttgccgtc tgaatttgac ctgagcgcat ttttacgcgc cggagaaaac cgcctcgcgg
      661 tgatggtgct gcgttggagt gacggcagtt atctggaaga tcaggatatg tggcggatga
      721 gcggcatttt ccgtgacgtg tcgttgctgc ataaaccgac tacacaaatc agcgatttcc
      781 atgttgccac tcgctttaat gatgatttca gccgcgctgt actggaggct gaagttcaga
      841 tgtgcggcga gttgcgtgac tacctacggg taacagtttc tttatggcag ggtgaaacgc
      901 aggtcgccag cggcaccgcg cctttcggcg gtgaaattat cgatgagcgt ggtggttatg
      961 ccgatcgcgt cacactacgt ctgaacgtcg aaaacccgaa actgtggagc gccgaaatcc
     1021 cgaatctcta tcgtgcggtg gttgaactgc acaccgccga cggcacgctg attgaagcag
     1081 aagcctgcga tgtcggtttc cgcgaggtgc ggattgaaaa tggtctgctg ctgctgaacg
     1141 gcaagccgtt gctgattcga ggcgttaacc gtcacgagca tcatcctctg catggtcagg
     1201 tcatggatga gcagacgatg gtgcaggata tcctgctgat gaagcagaac aactttaacg
     1261 ccgtgcgctg ttcgcattat ccgaaccatc cgctgtggta cacgctgtgc gaccgctacg
     1321 gcctgtatgt ggtggatgaa gccaatattg aaacccacgg catggtgcca atgaatcgtc
     1381 tgaccgatga tccgcgctgg ctaccggcga tgagcgaacg cgtaacgcga atggtgcagc
     1441 gcgatcgtaa tcacccgagt gtgatcatct ggtcgctggg gaatgaatca ggccacggcg
     1501 ctaatcacga cgcgctgtat cgctggatca aatctgtcga tccttcccgc ccggtgcagt
     1561 atgaaggcgg cggagccgac accacggcca ccgatattat ttgcccgatg tacgcgcgcg
     1621 tggatgaaga ccagcccttc ccggctgtgc cgaaatggtc catcaaaaaa tggctttcgc
     1681 tacctggaga gacgcgcccg ctgatccttt gcgaatacgc ccacgcgatg ggtaacagtc
     1741 ttggcggttt cgctaaatac tggcaggcgt ttcgtcagta tccccgttta cagggcggct
     1801 tcgtctggga ctgggtggat cagtcgctga ttaaatatga tgaaaacggc aacccgtggt
     1861 cggcttacgg cggtgatttt ggcgatacgc cgaacgatcg ccagttctgt atgaacggtc
     1921 tggtctttgc cgaccgcacg ccgcatccag cgctgacgga agcaaaacac cagcagcagt
     1981 ttttccagtt ccgtttatcc gggcaaacca tcgaagtgac cagcgaatac ctgttccgtc
     2041 atagcgataa cgagctgctg cactggatgg tggcgctgga tggtaagccg ctggcaagcg
     2101 gtgaagtgcc tctggatgtc gctccacaag gtaaacagtt gattgaactg cctgaactac
     2161 cgcagccgga gagcgccggg caactctggc tcacagtacg cgtagtgcaa ccgaacgcga
     2221 ccgcatggtc agaagccggg cacatcagcg cctggcagca gtggcgtctg gcggaaaacc
     2281 tcagtgtgac gctccccgcc gcgtcccacg ccatcccgca tctgaccacc agcgaaatgg
     2341 atttttgcat cgagctgggt aataagcgtt ggcaatttaa ccgccagtca ggctttcttt
     2401 cacagatgtg gattggcgat aaaaaacaac tgctgacgcc gctgcgcgat cagttcaccc
     2461 gtgcaccgct ggataacgac attggcgtaa gtgaagcgac ccgcattgac cctaacgcct
     2521 gggtcgaacg ctggaaggcg gcgggccatt accaggccga agcagcgttg ttgcagtgca
     2581 cggcagatac acttgctgat gcggtgctga ttacgaccgc tcacgcgtgg cagcatcagg
     2641 ggaaaacctt atttatcagc cggaaaacct accggattga tggtagtggt caaatggcga
     2701 ttaccgttga tgttgaagtg gcgagcgata caccgcatcc ggcgcggatt ggcctgaact
     2761 gccagctggc gcaggtagca gagcgggtaa actggctcgg attagggccg caagaaaact
     2821 atcccgaccg ccttactgcc gcctgttttg accgctggga tctgccattg tcagacatgt
     2881 ataccccgta cgtcttcccg agcgaaaacg gtctgcgctg cgggacgcgc gaattgaatt
     2941 atggcccaca ccagtggcgc ggcgacttcc agttcaacat cagccgctac agtcaacagc
     3001 aactgatgga aaccagccat cgccatctgc tgcacgcgga agaaggcaca tggctgaata
     3061 tcgacggttt ccatatgggg attggtggcg acgactcctg gagcccgtca gtatcggcgg
     3121 agttccagct gagcgccggt cgctaccatt accagttggt ctggtgtcaa aaataataat
     3181 aaccgggcag gccatgtctg cccgtatttc gcgtaaggaa atccattatg tactatttaa
     3241 aatcaagctc cttgcaggcc actagtcttg gactcctgtt gatagatcca gtaatgacct
     3301 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg
     3361 agaatccagg ggtccccaat aattacgatt taaatttgtg tctcaaaatc tctgatgtta
     3421 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     3481 taatacaagg ggtgttatga gccatattca gcgtgaaacg agctgtagcc gtccgcgtct
     3541 gaacagcaac atggatgcgg atctgtatgg ctataaatgg gcgcgtgata acgtgggtca
     3601 gagcggcgcg accatttatc gtctgtatgg caaaccggat gcgccggaac tgtttctgaa
     3661 acatggcaaa ggcagcgtgg cgaacgatgt gaccgatgaa atggtgcgtc tgaactggct
     3721 gaccgaattt atgccgctgc cgaccattaa acattttatt cgcaccccgg atgatgcgtg
     3781 gctgctgacc accgcgattc cgggcaaaac cgcgtttcag gtgctggaag aatatccgga
     3841 tagcggcgaa aacattgtgg atgcgctggc cgtgtttctg cgtcgtctgc atagcattcc
     3901 ggtgtgcaac tgcccgttta acagcgatcg tgtgtttcgt ctggcccagg cgcagagccg
     3961 tatgaacaac ggcctggtgg atgcgagcga ttttgatgat gaacgtaacg gctggccggt
     4021 ggaacaggtg tggaaagaaa tgcataaact gctgccgttt agcccggata gcgtggtgac
     4081 ccacggcgat tttagcctgg ataacctgat tttcgatgaa ggcaaactga ttggctgcat
     4141 tgatgtgggc cgtgtgggca ttgcggatcg ttatcaggat ctggccattc tgtggaactg
     4201 cctgggcgaa tttagcccga gcctgcaaaa acgtctgttt cagaaatatg gcattgataa
     4261 tccggatatg aacaaactgc aatttcatct gatgctggat gaatttttct aataattaat
     4321 tggaccgcgg tccgcgcgtt gtccttttcc gctgcataac cctgcttcgg ggtcattata
     4381 gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt
     4441 cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc
     4501 ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac
     4561 gggaatcctg ctctgcgagg ctggccgtag gccggccgcg atgcaggtgg ctgctgaacc
     4621 cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt catcattgac
     4681 ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc cgacctgctc
     4741 gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg tttccagctt
     4801 gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga
     4861 cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa acagcacgac
     4921 gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt ccaggacgcg
     4981 gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg tgaagcccat
     5041 cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc ggccattgat
     5101 cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct cgccgatagg
     5161 ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt cggcccgcag
     5221 ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga ccttgttttg
     5281 cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc
     5341 gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg aaagctgcat
     5401 ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggctt cgctgacctg
     5461 ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc
     5521 gatggtcatc gacttcgcca aacctgccgc ctcctgttcg agacgacgcg aacgctccac
     5581 ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc gctcgatctt
     5641 ggccgtagct tgctggacta tcgagccgac ggactggaag gtttcgcggg gcgcacgcat
     5701 gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc
     5761 ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg ggattgcccc
     5821 ggaattaatt ccccggatcg atccgtcgat cttgatcccc tgcgccatca gatccttggc
     5881 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     5941 gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta
     6001 agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc
     6061 cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtggct
     6121 gccatttttg gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg ggatgggagg
     6181 cccgcgttag cgggccggga gggttcgaga agggggggca ccccccttcg gcgtgcgcgg
     6241 tcacgcgcac agggcgcagc cctggttaaa aacaaggttt ataaatattg gtttaaaagc
     6301 aggttaaaag acaggttagc ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga
     6361 ttttctgcct gtggacagcc cctcaaatgt caataggtgc gcccctcatc tgtcagcact
     6421 ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg
     6481 tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta
     6541 aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg
     6601 agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc
     6661 cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc
     6721 ctacatggct ctgctgtagt gagtgggttg cgctccggca gcggtcctga tcccccgcag
     6781 aaaaaaagga tctcaagaag atcctttgat cttttctacg gcgcgcccag ctgtctaggg
     6841 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     6901 ccagtctttc gactgagcct ttcgttttat ttgatgcct
//
