LOCUS       Exported                6607 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pVLG6, complete sequence.
ACCESSION   JF812998
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6607)
  AUTHORS   Zeigler DR.
  TITLE     Sequence of pVLG6, a shuttle vector for constructing GFP fusions in 
            Bacillus megaterium and other Gram-positive bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6607)
  AUTHORS   Zeigler DR.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-APR-2011) Bacillus Genetic Stock Center, The Ohio 
            State University, 484 W 12th Ave, Columbus, OH 43210, USA
REFERENCE   3  (bases 1 to 6607)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6607
                     /organism="Cloning vector pVLG6"
                     /mol_type="other DNA"
                     /note="plasmid depositied in Bacillus Genetic Stock Center 
                     by V. Lukyanchuk and P. S. Vary, Northern Illinois 
                     University"
                     /db_xref="taxon:1086586"
     primer_bind     18..34
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             61..777
                     /codon_start=1
                     /product="GFPmut1"
                     /label=GFPmut1
                     /note="green fluorescent protein; enhanced fluorescence GFP
                     at 488 nm"
                     /protein_id="AEP13835.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLSPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(792..808)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1202..1387
                     /codon_start=1
                     /gene="cop"
                     /product="Cop"
                     /label=cop
                     /note="copy control regulatory protein for Gram-positive 
                     replication"
                     /protein_id="AEP13839.1"
                     /translation="MLGGTVMVVDRKEEKKVAVTLRLTTEENEILNRIKEKYNISKSDA
                     TGILIKKYAKEEYGAF"
     gene            1202..1387
                     /gene="cop"
                     /label=cop
     CDS             1468..2067
                     /codon_start=1
                     /gene="repF"
                     /product="RepF"
                     /label=repF
                     /note="initiation protein for Gram-positive replication"
                     /protein_id="AEP13838.1"
                     /translation="MSENVIKETENKKNSRGRNWTFVLYPESAKAEWLEYLKELHIQFV
                     VSPLHDRDTDTEGRMKKEHYHILVMYEGNKSYEQIKIITEELNATIPQIAGSVKGLVRY
                     MLHMDDPNKFKYQKEDMIVYGGVDVDELLKKTTTDRYKLIKEMIEFIDEQGIVEFKSLM
                     DYAMKFKFDDWFPLLCDNSAYVIQEYIKSNRYKSDR"
     gene            1468..2067
                     /gene="repF"
                     /label=repF
     promoter        complement(2703..2721)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2728..2744)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2885..3340)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3367..3471
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3472..4332
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /note="beta-lactamase; ampicillin resistance"
                     /protein_id="AEP13837.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            3472..4332
                     /gene="bla"
                     /label=bla
     rep_origin      4503..5091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5455..6105)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="chloramphenicol resistance"
                     /protein_id="AEP13836.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            complement(5455..6105)
                     /gene="cat"
                     /label=cat
     protein_bind    6443..6464
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6479..6509
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6517..6533
                     /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)."
     primer_bind     6541..6557
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6578..6596
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"