LOCUS       40924_39268        5250 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSEVA224, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5250)
  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 5250)
  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 5250)
  AUTHORS   Silva-Rocha R, Martinez-Garcia E, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-SEP-2014) Systems Biology Program, CNB-CSIC, Darwin 3,
            Madrid, Madrid 28029, Spain
REFERENCE   4  (bases 1 to 5250)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5250)
  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"; journalName: "Submitted 
            (30-SEP-2014) Systems Biology Program, CNB-CSIC, Darwin 3, Madrid, 
            Madrid 28029, Spain"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
            On Sep 30, 2014 this sequence version replaced KC847291.1.
FEATURES             Location/Qualifiers
     source          1..5250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..88
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             89..1168
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        1403..1432
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    1440..1456
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     misc_feature    1483..1539
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1548..1571)
                     /label=R24
                     /note="R24"
     terminator      1582..1676
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1808..2620
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            2790..2898
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2962..4107)
                     /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(4500..5031)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(5158..5244)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaatt gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc
       61 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
      121 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
      181 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
      241 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
      301 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
      361 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
      421 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
      481 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
      541 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
      601 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
      661 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
      721 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
      781 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
      841 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
      901 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
      961 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     1021 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     1081 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     1141 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagcgcg
     1201 aattgatctg gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca
     1261 ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt
     1321 cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg
     1381 gcaaatattc tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat
     1441 tgtgagcgga taacaatttc acaccctagg ccgcggccgc gcgaattcga gctcggtacc
     1501 cggggatcct ctagagtcga cctgcaggca tgcaagcttg cggccgcgtc gtgactggga
     1561 aaaccctggc gactagtctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
     1621 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccag
     1681 gggtccccaa taattacgat ttaaatttgt gtctcaaaat ctctgatgtt acattgcaca
     1741 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1801 gggtgttatg agccatattc agcgtgaaac gagctgtagc cgtccgcgtc tgaacagcaa
     1861 catggatgcg gatctgtatg gctataaatg ggcgcgtgat aacgtgggtc agagcggcgc
     1921 gaccatttat cgtctgtatg gcaaaccgga tgcgccggaa ctgtttctga aacatggcaa
     1981 aggcagcgtg gcgaacgatg tgaccgatga aatggtgcgt ctgaactggc tgaccgaatt
     2041 tatgccgctg ccgaccatta aacattttat tcgcaccccg gatgatgcgt ggctgctgac
     2101 caccgcgatt ccgggcaaaa ccgcgtttca ggtgctggaa gaatatccgg atagcggcga
     2161 aaacattgtg gatgcgctgg ccgtgtttct gcgtcgtctg catagcattc cggtgtgcaa
     2221 ctgcccgttt aacagcgatc gtgtgtttcg tctggcccag gcgcagagcc gtatgaacaa
     2281 cggcctggtg gatgcgagcg attttgatga tgaacgtaac ggctggccgg tggaacaggt
     2341 gtggaaagaa atgcataaac tgctgccgtt tagcccggat agcgtggtga cccacggcga
     2401 ttttagcctg gataacctga ttttcgatga aggcaaactg attggctgca ttgatgtggg
     2461 ccgtgtgggc attgcggatc gttatcagga tctggccatt ctgtggaact gcctgggcga
     2521 atttagcccg agcctgcaaa aacgtctgtt tcagaaatat ggcattgata atccggatat
     2581 gaacaaactg caatttcatc tgatgctgga tgaatttttc taataattaa ttggaccgcg
     2641 gtccgcgcgt tgtccttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt
     2701 tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac
     2761 tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg
     2821 agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct
     2881 gctctgcgag gctggccgta ggccggccgc gatgcaggtg gctgctgaac ccccagccgg
     2941 aactgacccc acaaggccct agcgtttgca atgcaccagg tcatcattga cccaggcgtg
     3001 ttccaccagg ccgctgcctc gcaactcttc gcaggcttcg ccgacctgct cgcgccactt
     3061 cttcacgcgg gtggaatccg atccgcacat gaggcggaag gtttccagct tgagcgggta
     3121 cggctcccgg tgcgagctga aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg
     3181 gtacttctcc catatgaatt tcgtgtagtg gtcgccagca aacagcacga cgatttcctc
     3241 gtcgatcagg acctggcaac gggacgtttt cttgccacgg tccaggacgc ggaagcggtg
     3301 cagcagcgac accgattcca ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc
     3361 ctgtaggcgc gacaggcatt cctcggcctt cgtgtaatac cggccattga tcgaccagcc
     3421 caggtcctgg caaagctcgt agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt
     3481 cgcgtactcc aacacctgct gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc
     3541 ggtgtaggtg atcttcacgt ccttgttgac gtggaaaatg accttgtttt gcagcgcctc
     3601 gcgcgggatt ttcttgttgc gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat
     3661 cgctcgcatc gtgtccggcc acggcgcaat atcgaacaag gaaagctgca tttccttgat
     3721 ctgctgcttc gtgtgtttca gcaacgcggc ctgcttggct tcgctgacct gttttgccag
     3781 gtcctcgccg gcggtttttc gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat
     3841 cgacttcgcc aaacctgccg cctcctgttc gagacgacgc gaacgctcca cggcggccga
     3901 tggcgcgggc agggcagggg gagccagttg cacgctgtcg cgctcgatct tggccgtagc
     3961 ttgctggact atcgagccga cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg
     4021 gcttgcgatg gtttcggcat cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta
     4081 tgccttccgg tcaaacgtcc gattcattca ccctccttgc gggattgccc cggaattaat
     4141 tccccggatc gatccgtcga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa
     4201 gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
     4261 tccggttcgc ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg
     4321 caagctacct gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg
     4381 acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtggc tgccattttt
     4441 ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg gggatgggag gcccgcgtta
     4501 gcgggccggg agggttcgag aagggggggc accccccttc ggcgtgcgcg gtcacgcgca
     4561 cagggcgcag ccctggttaa aaacaaggtt tataaatatt ggtttaaaag caggttaaaa
     4621 gacaggttag cggtggccga aaaacgggcg gaaacccttg caaatgctgg attttctgcc
     4681 tgtggacagc ccctcaaatg tcaataggtg cgcccctcat ctgtcagcac tctgcccctc
     4741 aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg cccctcaagt gtcaataccg
     4801 cagggcactt atccccaggc ttgtccacat catctgtggg aaactcgcgt aaaatcaggc
     4861 gttttcgccg atttgcgagg ctggccagct ccacgtcgcc ggccgaaatc gagcctgccc
     4921 ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa gtgtcaacgt ccgcccctca
     4981 tctgtcagtg agggccaagt tttccgcgag gtatccacaa cgccggcggc cctacatggc
     5041 tctgctgtag tgagtgggtt gcgctccggc agcggtcctg atcccccgca gaaaaaaagg
     5101 atctcaagaa gatcctttga tcttttctac ggcgcgccca gctgtctagg gcggcggatt
     5161 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt
     5221 cgactgagcc tttcgtttta tttgatgcct
//
