LOCUS       Exported                7616 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJAC91, complete sequence.
ACCESSION   KF322087
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7616)
  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 7616)
  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 7616)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7616
                     /organism="Cloning vector pJAC91"
                     /mol_type="other DNA"
                     /db_xref="taxon:1403159"
     misc_feature    38..501
                     /label=HIS3 5UTR
                     /note="HIS3 5UTR"
     promoter        314..501
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     misc_feature    502..1915
                     /label=CG HIS3 w promoter and terminator
                     /note="CG HIS3 w promoter and terminator"
     regulatory      1928..2189
                     /regulatory_class="promoter"
                     /note="PCyc1 promoter"
     CDS             2175..3104
                     /codon_start=1
                     /product="Gal4AD-linker-N-Ada"
                     /label=Gal4AD-linker-N-Ada
                     /protein_id="AGW27650.1"
                     /translation="MTELEGAANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALS
                     QPIASSNVHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDV
                     YNYLFDDEDTPPNPKKEGSGSGSGSKKATCLTDDQRWQSVLARDPNADGEFVFAVRTTG
                     IFCRPSCRARHALRENVSFYANASEALAAGFRPCKRCQPEKANAQQHRLDKITHACRLL
                     EQETPVTLEALADQVAMSPFHLHRLFKATTGMTPKAWQQAWRARRLRESLAKGESVTTS
                     ILNAGFPDSSSYYRKADETLGMTAKQFR"
     CDS             2196..2537
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     regulatory      3105..3799
                     /regulatory_class="terminator"
                     /note="C. albicans TadH1 terminator"
     misc_feature    3800..4118
                     /label=HIS3 3UTR
                     /note="HIS3 3UTR"
     promoter        complement(4164..4182)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4189..4205)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4346..4801
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      4351..4791
                     /label=F1 ori
                     /note="F1 ori"
     promoter        5439..5543
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5544..6404
                     /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             5742..6404
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AGW27651.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     rep_origin      6499..7181
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     rep_origin      6575..7163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7451..7472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7487..7517
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7525..7541
                     /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     7549..7565
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7586..7604
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"