LOCUS       Exported                7589 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector PJAC93, complete sequence.
ACCESSION   KF322089
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7589)
  AUTHORS   Moser F, Horwitz A, Chen J, Lim W, Voigt CA.
  TITLE     Genetic sensor for strong methylating compounds
  JOURNAL   ACS Synth Biol 2 (10), 614-624 (2013)
  PUBMED    24032656
REFERENCE   2  (bases 1 to 7589)
  AUTHORS   Moser F, Horwitz A, Chen J, Lim WA, Voigt CA.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-JUL-2013) Biological Engineering, MIT, 500 Tech 
            Square, Rm 209D, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 7589)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7589
                     /organism="Cloning vector PJAC93"
                     /mol_type="other DNA"
                     /db_xref="taxon:1403154"
     misc_feature    434..1934
                     /label=CG TRP1 w promoter and terminator
                     /note="CG TRP1 w promoter and terminator"
     misc_feature    1941..2027
                     /label=3x Ada operator
                     /note="3x Ada operator"
     regulatory      2028..2274
                     /regulatory_class="promoter"
                     /note="PCyc1 promoter"
     CDS             2275..3012
                     /codon_start=1
                     /product="EGFP"
                     /label=EGFP
                     /protein_id="AGW27654.1"
                     /translation="MTELETSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYG
                     KLTLKFICTTGKLPVPWPTLVTTLCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIF
                     FKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQK
                     NGIKVNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDH
                     MVLLEFVTAAGITHGMDELYNGS"
     CDS             2293..3000
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELY"
     regulatory      3045..3739
                     /regulatory_class="terminator"
                     /note="C. albicans TadH1 terminator"
     promoter        complement(4137..4155)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4162..4178)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4319..4774
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      4324..4764
                     /label=F1 ori
                     /note="F1 ori"
     promoter        5412..5516
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5517..6377
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             5715..6377
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AGW27655.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     rep_origin      6472..7154
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     rep_origin      6548..7136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7424..7445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7460..7490
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7498..7514
                     /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     7522..7538
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7559..7577
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"