LOCUS       Exported                7420 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pYFPamy, complete sequence.
ACCESSION   HM204937
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7420)
  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 7420)
  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 7420)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7420
                     /organism="Cloning vector pYFPamy"
                     /lab_host="Bacillus subtilis"
                     /mol_type="other DNA"
                     /db_xref="taxon:881626"
     misc_recomb     115..651
                     /label=amyE front
                     /note="amyE front"
     gene            complement(658..1532)
                     /gene="iyfp"
                     /label=iyfp
     primer_bind     685..701
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        708..726
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     759..775
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(777..1532)
                     /codon_start=1
                     /gene="iyfp"
                     /product="IYFP"
                     /label=iyfp
                     /protein_id="ADM95149.1"
                     /translation="MDSIEKVSEFATMVSKGEELFTGVVPILVELDGDVNGHKFSVSGE
                     GEGDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEG
                     YVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNV
                     YIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSK
                     DPNEKRDHMVLLEFVTAAGITLGMDELYK"
     CDS             complement(777..1496)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      1493..1502
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    complement(1527..1562)
                     /label=LIC site
                     /note="LIC site"
     CDS             1721..2371
                     /codon_start=1
                     /product="chloramphenicol resistance determinant"
                     /label=chloramphenicol resistance determinant
                     /protein_id="ADM95150.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     misc_recomb     2736..3772
                     /label=amyE back
                     /note="amyE back"
     CDS             4972..5163
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    5265..5405
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5570..6189)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      complement(5591..6179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6350..7210)
                     /codon_start=1
                     /product="ampicillin resistance determinant"
                     /label=ampicillin resistance determinant
                     /protein_id="ADM95151.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7211..7315)
                     /gene="bla"
                     /label=AmpR promoter