LOCUS       Exported                6267 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJFR4, complete sequence.
ACCESSION   KX011467
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6267)
  AUTHORS   Fernandez-Rodriguez J, Moser F, Voigt CA.
  TITLE     Multichromatic Bacterial Photography
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6267)
  AUTHORS   Fernandez-Rodriguez J, Moser F, Voigt CA.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAR-2016) Biological Engineering, MIT, 500 Tech 
            Square, Rm 140, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 6267)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6267
                     /organism="Cloning vector pJFR4"
                     /mol_type="other DNA"
                     /db_xref="taxon:1978235"
     regulatory      253..275
                     /regulatory_class="promoter"
                     /note="Promoter T3 (T7 RNAP)"
     CDS             311..1003
                     /codon_start=1
                     /product="BFP"
                     /label=BFP
                     /protein_id="ARG47680.1"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGH"
     misc_feature    1022..1171
                     /label=Terminator DT5
                     /note="Terminator DT5"
     regulatory      1212..1234
                     /regulatory_class="promoter"
                     /note="Promoter K1F (T7 RNAP)"
     misc_feature    1276..1938
                     /label=similar to mRFP1
                     /note="similar to mRFP1"
     misc_feature    1945..1971
                     /label=similar to FLAG tag
                     /note="similar to FLAG tag"
     CDS             1945..1968
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1982..2140
                     /label=Terminator DT16
                     /note="Terminator DT16"
     terminator      2176..2223
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    complement(2176..2222)
                     /label=Terminator T7
                     /note="Terminator T7"
     CDS             complement(2244..2960)
                     /codon_start=1
                     /product="sfGFP"
                     /label=sfGFP
                     /protein_id="ARG47681.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(2247..2960)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      2996..3018
                     /regulatory_class="promoter"
                     /note="Promoter CCG (T7 RNAP)"
     CDS             complement(3570..4520)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3573..4520
                     /label=pSC101 origin
                     /note="pSC101 origin"
     rep_origin      4568..4790
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     misc_feature    5275..5356
                     /label=Terminator T0
                     /note="Terminator T0"
     terminator      5282..5376
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5407..6267)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGTRVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(5407..6069)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ARG47682.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     regulatory      complement(6070..6267)
                     /regulatory_class="promoter"
                     /note="Promoter AmpR"