LOCUS       40924_10511        6032 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCFPbglS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6032)
  AUTHORS   Bisicchia P, Botella E, Devine KM.
  TITLE     Suite of novel vectors for ectopic insertion of GFP, CFP and IYFP 
            transcriptional fusions in single copy at the amyE and bglS loci in 
            Bacillus subtilis
  JOURNAL   Plasmid 64 (3), 143-149 (2010)
  PUBMED    20600285
REFERENCE   2  (bases 1 to 6032)
  AUTHORS   Bisicchia P.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAY-2010) Trinity College Dublin, 13 Meades Terrace, 
            Dublin 2, Ireland
REFERENCE   3  (bases 1 to 6032)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6032)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2010"; volume: "64"; issue: "3"; pages: "143-149"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-MAY-2010) Trinity College Dublin, 13 Meades Terrace, Dublin 2, 
            Ireland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     695..1121
                     /label=bglS front
                     /note="bglS front"
     misc_feature    1134..1169
                     /label=LIC site
                     /note="LIC site"
     CDS             1170..1886
                     /codon_start=1
                     /label=mECFP
                     /note="enhanced CFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2446..3237
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     primer_bind     complement(3397..3413)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     3416..3813
                     /label=bglS back
                     /note="bglS back"
     promoter        complement(3847..3865)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3886..3902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3910..3926)
                     /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)."
     promoter        complement(3934..3964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3979..4000)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4288..4876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5050..5907)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5908..6012)
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactagtgg atccgcttca ctattatcgg ttcgtcaccc
      721 acttaaagtt tttcgcgcag cgtctattta acggcactca catggaaagc caggacgatt
      781 ttttgctgga tacagtgaaa gaaaagtatc atcgcgcgta tgaatgcacg aagaaaatcc
      841 aaacctacat tgagcgggag tatgagcaca agctcacaag tgacgagctg ctgtatttaa
      901 ccattcacat agaaagggta gttaaacaag cataatgaga gcgctgacat ttgtgtttcc
      961 ttgtgttcac tttttcttac attcacatat gaaaatggta ggattgttac tgataaagca
     1021 ggcaaaacct aaattgcaat gagtgcggat catctctgtc tgtgctgatg gtaatttagg
     1081 tttttatttt tttcagaggg aagatgatga tagttacagg actgcaggaa ttcttttacc
     1141 gcgggctttc ccgggaagga ggaactacta tggtttcaaa aggcgaagaa ctgtttacgg
     1201 gagtagtgcc gattctggtc gaattagatg gcgatgtcaa tggccataaa ttttctgtct
     1261 caggcgaggg agaaggagat gctacgtacg gcaaactgac gcttaagttc atctgcacaa
     1321 caggaaagtt acctgtccca tggcctacat tagtcacgac acttacgtgg ggagtacagt
     1381 gcttcagcag atacccggat cacatgaaac agcacgattt cttcaaatca gccatgccgg
     1441 aaggctatgt gcaagaacgc acgatttttt tcaaggacga cggaaattat aaaacacgcg
     1501 cggaagtcaa atttgaaggc gacacgcttg tcaatagaat cgaactgaaa ggaattgact
     1561 ttaaggagga tggcaacatc ttaggacaca aactggaata taattacatc tctcacaatg
     1621 tttatatcac agcagataag caaaaaaacg gaatcaaggc taatttcaaa atccgccata
     1681 acattgagga cggaagcgtt caattagcag atcattacca gcagaataca ccgattggag
     1741 atggaccggt tcttcttcct gacaaccatt acttaagcac gcaatctaag ctgtcaaaag
     1801 accctaacga aaagcgcgat cacatggttc tgttagagtt tgtcacagct gcgggcatta
     1861 cacttggcat ggacgaactt tataagtaag aattcgatat caaagcttat cgataaaccc
     1921 agcgaaccat ttgaggtgat aggtaagatt ataccgaggt atgaaaacga gaattggacc
     1981 tttacagaat tactctatga agcgccatat ttaaaaagct accaagacga agaggatgaa
     2041 gaggatgagg aggcagattg ccttgaatat attgacaata ctgataagat aatatatctt
     2101 ttatatagaa gatatcgccg tatgtaagga tttcaggggg caaggcatag gcagcgcgct
     2161 tatcaatata tctatagaat gggcaaagca taaaaacttg catggactaa tgcttgaaac
     2221 ccaggacaat aaccttatag cttgtaaatt ctatcataat tgtggtttca aaatcggctc
     2281 cgtcgatact atgttatacg ccaactttca aaacaacttt gaaaaagctg ttttctggta
     2341 tttaaggttt tagaatgcaa ggaacagtga attggagttc gtcttgttat aattagcttc
     2401 ttggggtatc tttaaatact gtagaaaaga ggaaggaaat aataaatggc taaaatgaga
     2461 atatcaccgg aattgaaaaa actgatcgaa aaataccgct gcgtaaaaga tacggaagga
     2521 atgtctcctg ctaaggtata taagctggtg ggagaaaatg aaaacctata tttaaaaatg
     2581 acggacagcc ggtataaagg gaccacctat gatgtggaac gggaaaagga catgatgcta
     2641 tggctggaag gaaagctgcc tgttccaaag gtcctgcact ttgaacggca tgatggctgg
     2701 agcaatctgc tcatgagtga ggccgatggc gtcctttgct cggaagagta tgaagatgaa
     2761 caaagccctg aaaagattat cgagctgtat gcggagtgca tcaggctctt tcactccatc
     2821 gacatatcgg attgtcccta tacgaatagc ttagacagcc gcttagccga attggattac
     2881 ttactgaata acgatctggc cgatgtggat tgcgaaaact gggaagaaga cactccattt
     2941 aaagatccgc gcgagctgta tgatttttta aagacggaaa agcccgaaga ggaacttgtc
     3001 ttttcccacg gcgacctggg agacagcaac atctttgtga aagatggcaa agtaagtggc
     3061 tttattgatc ttgggagaag cggcagggcg gacaagtggt atgacattgc cttctgcgtc
     3121 cggtcgatca gggaggatat cggggaagaa cagtatgtcg agctattttt tgacttactg
     3181 gggatcaagc ctgattggga gaaaataaaa tattatattt tactggatga attgttttag
     3241 tacctagatt tagatgtcta aaaagcttta actacaagct ttttagacat ctaatctttt
     3301 ctgaagtaca tccgcaactg tccatactct gatgttttat atcttttcta aaagttcgct
     3361 agataggggt cccgagcgcc tacgaggaat ttgtatcgat accgtcgacc tcgagcttca
     3421 cttacacagg tccaacagat ggaactcctt gggatgagat tgatatcgaa tttttaggaa
     3481 aagacacaac gaaggttcaa tttaactatt atacaaatgg tgcaggaaac catgagaaga
     3541 ttgttgatct cgggtttgat gcagccaatg cctatcatac gtatgcattc gattggcagc
     3601 caaactctat taaatggtat gtcgacgggc aattaaaaca tactgcaaca aaccaaattc
     3661 cgacaacacc tggaaagatc atgatgaact tgtggaatgg cacgggtgtc gatgaatggc
     3721 ttggctccta caatggtgta aatccgctat acgctcatta tgactgggtg cgctatacaa
     3781 aaaaataatg ccaaatgtga aagagcctgc tgcctcgagg gggggcccgg tacccagctt
     3841 ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat agctgtttcc
     3901 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3961 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4021 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4081 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4141 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4201 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4261 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4321 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4381 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4441 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4501 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4561 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4621 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4681 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4741 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4801 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4861 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4921 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4981 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5041 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5101 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5161 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5221 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5281 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5341 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5401 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5461 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5521 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5581 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5641 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5701 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5761 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5821 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5881 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5941 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6001 aggggttccg cgcacatttc cccgaaaagt gc
//
