LOCUS       40924_48113        6181 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pYFPbglS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6181)
  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 6181)
  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 6181)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6181)
  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..6181
                     /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             1206..1922
                     /codon_start=1
                     /label=EYFP
                     /note="enhanced YFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1927..1943)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1976..1994)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2001..2017)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2595..3386
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     primer_bind     complement(3546..3562)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     3565..3962
                     /label=bglS back
                     /note="bglS back"
     promoter        complement(3996..4014)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4035..4051)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4059..4075)
                     /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(4083..4113)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4128..4149)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4437..5025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5199..6056)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6057..6161)
                     /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 tggattcaat agaaaaggta agcgaatttg
     1201 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     1261 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1321 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1381 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     1441 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1501 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1561 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1621 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1681 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1741 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1801 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1861 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1921 agtaaagatc cactagttct agagcggccg ccaccgcggt ggagctccaa ttcgccctat
     1981 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccga
     2041 attcgatatc aaagcttatc gataaaccca gcgaaccatt tgaggtgata ggtaagatta
     2101 taccgaggta tgaaaacgag aattggacct ttacagaatt actctatgaa gcgccatatt
     2161 taaaaagcta ccaagacgaa gaggatgaag aggatgagga ggcagattgc cttgaatata
     2221 ttgacaatac tgataagata atatatcttt tatatagaag atatcgccgt atgtaaggat
     2281 ttcagggggc aaggcatagg cagcgcgctt atcaatatat ctatagaatg ggcaaagcat
     2341 aaaaacttgc atggactaat gcttgaaacc caggacaata accttatagc ttgtaaattc
     2401 tatcataatt gtggtttcaa aatcggctcc gtcgatacta tgttatacgc caactttcaa
     2461 aacaactttg aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa
     2521 ttggagttcg tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag
     2581 gaaggaaata ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa
     2641 aataccgctg cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg
     2701 gagaaaatga aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg
     2761 atgtggaacg ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg
     2821 tcctgcactt tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg
     2881 tcctttgctc ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg
     2941 cggagtgcat caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct
     3001 tagacagccg cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt
     3061 gcgaaaactg ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa
     3121 agacggaaaa gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca
     3181 tctttgtgaa agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg
     3241 acaagtggta tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac
     3301 agtatgtcga gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat
     3361 attatatttt actggatgaa ttgttttagt acctagattt agatgtctaa aaagctttaa
     3421 ctacaagctt tttagacatc taatcttttc tgaagtacat ccgcaactgt ccatactctg
     3481 atgttttata tcttttctaa aagttcgcta gataggggtc ccgagcgcct acgaggaatt
     3541 tgtatcgata ccgtcgacct cgagcttcac ttacacaggt ccaacagatg gaactccttg
     3601 ggatgagatt gatatcgaat ttttaggaaa agacacaacg aaggttcaat ttaactatta
     3661 tacaaatggt gcaggaaacc atgagaagat tgttgatctc gggtttgatg cagccaatgc
     3721 ctatcatacg tatgcattcg attggcagcc aaactctatt aaatggtatg tcgacgggca
     3781 attaaaacat actgcaacaa accaaattcc gacaacacct ggaaagatca tgatgaactt
     3841 gtggaatggc acgggtgtcg atgaatggct tggctcctac aatggtgtaa atccgctata
     3901 cgctcattat gactgggtgc gctatacaaa aaaataatgc caaatgtgaa agagcctgct
     3961 gcctcgaggg ggggcccggt acccagcttt tgttcccttt agtgagggtt aatttcgagc
     4021 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4081 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4141 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4201 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4261 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4321 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4381 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4441 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4501 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4561 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4621 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4681 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4741 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4801 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4861 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4921 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4981 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5041 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5101 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5161 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5221 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5281 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5341 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5401 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5461 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5521 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5581 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5641 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5701 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5761 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5821 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5881 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5941 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6001 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6061 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6121 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6181 c
//
