LOCUS       Exported                5042 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCZ-NAT, complete sequence.
ACCESSION   JN580989
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5042)
  AUTHORS   Levy SF, Ziv N, Siegal ML.
  TITLE     Bet hedging in yeast by heterogeneous, age-correlated expression of 
            a stress protectant
  JOURNAL   PLoS Biol. 10 (5), E1001325 (2012)
  PUBMED    22589700
REFERENCE   2  (bases 1 to 5042)
  AUTHORS   Levy SF, Lyman CE, Ziv N, Siegal ML.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-AUG-2011) Genetics, Stanford University, S225 Grant 
            Building, Stanford University Medical School, 300 Pasteur Drive, 
            Stanford, CA 94305-5120, USA
REFERENCE   3  (bases 1 to 5042)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5042
                     /organism="Cloning vector pCZ-NAT"
                     /mol_type="other DNA"
                     /db_xref="taxon:1131568"
     regulatory      70..448
                     /regulatory_class="promoter"
                     /note="TEF promoter"
     promoter        105..448
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             449..1021
                     /codon_start=1
                     /gene="nat1"
                     /product="nourseothricin resistance"
                     /label=nat1
                     /protein_id="AEX28238.1"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     gene            449..1021
                     /gene="nat1"
                     /label=nat1
     protein_bind    1076..1092
                     /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     1100..1116
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1137..1155
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1206..1604
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     1608..1624
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1641..2351
                     /codon_start=1
                     /gene="mCherry"
                     /product="mCherry fluorophore"
                     /label=mCherry
                     /protein_id="AEX28237.1"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     gene            1641..2351
                     /gene="mCherry"
                     /label=mCherry
     terminator      2375..2572
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(2680..2698)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2956..3544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3715..4575)
                     /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"
     promoter        complement(4576..4680)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5026..2
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"