LOCUS       Exported                6181 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pYFPbglS, complete sequence.
ACCESSION   HM204936
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6181
                     /organism="Cloning vector pYFPbglS"
                     /lab_host="Bacillus subtilis"
                     /mol_type="other DNA"
                     /db_xref="taxon:881627"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(138..444)
                     /direction=LEFT
                     /label=f1(+) origin
                     /note="f1(+) origin"
     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"
     gene            1170..2045
                     /gene="iyfp"
                     /label=iyfp
     CDS             1170..1925
                     /codon_start=1
                     /gene="iyfp"
                     /product="IYFP"
                     /label=iyfp
                     /protein_id="ADM95146.1"
                     /translation="MDSIEKVSEFATMVSKGEELFTGVVPILVELDGDVNGHKFSVSGE
                     GEGDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEG
                     YVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNV
                     YIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSK
                     DPNEKRDHMVLLEFVTAAGITLGMDELYK"
     regulatory      1200..1209
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1206..1925
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /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..3389
                     /codon_start=1
                     /product="kanamycin resistance determinant"
                     /label=kanamycin resistance determinant
                     /protein_id="ADM95147.1"
                     /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    4059..4075
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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    4128..4149
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4422..5041)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      complement(4437..5025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5196..6056)
                     /codon_start=1
                     /product="ampicillin resistance determinant"
                     /label=ampicillin resistance determinant
                     /protein_id="ADM95148.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6057..6161)
                     /gene="bla"
                     /label=AmpR promoter