LOCUS       40924_14085        5042 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCZ-NAT, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5042)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Biol."; date: "2012"; volume: "10"; issue: "5"; pages: "E1001325"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-AUG-2011) Genetics, Stanford University, S225 Grant Building, 
            Stanford University Medical School, 300 Pasteur Drive, Stanford, CA 
            94305-5120, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        105..448
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             455..1018
                     /codon_start=1
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
                     /translation="TTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFTL
                     REVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRRL
                     TVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTLC
                     GLDTALYDGTASDGEQALYMSMPCP"
     protein_bind    1076..1092
                     /label=lac operator
                     /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        1205..1604
                     /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..2348
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     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(3718..4575)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4576..4680)
                     /label=AmpR promoter
     promoter        5026..5042
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gcttgccttg tccccgccgg gtcacccggc cagcgacatg gaggcccaga
      121 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
      181 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
      241 cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc
      301 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
      361 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
      421 ctcacatcac atccgaacat aaacaaccat gggtaccact cttgacgaca cggcttaccg
      481 gtaccgcacc agtgtcccgg gggacgccga ggccatcgag gcactggatg ggtccttcac
      541 caccgacacc gtcttccgcg tcaccgccac cggggacggc ttcaccctgc gggaggtgcc
      601 ggtggacccg cccctgacca aggtgttccc cgacgacgaa tcggacgacg aatcggacga
      661 cggggaggac ggcgacccgg actcccggac gttcgtcgcg tacggggacg acggcgacct
      721 ggcgggcttc gtggtcgtct cgtactccgg ctggaaccgc cggctgaccg tcgaggacat
      781 cgaggtcgcc ccggagcacc gggggcacgg ggtcgggcgc gcgttgatgg ggctcgcgac
      841 ggagttcgcc cgcgagcggg gcgccgggca cctctggctg gaggtcacca acgtcaacgc
      901 accggcgatc cacgcgtacc ggcggatggg gttcaccctc tgcggcctgg acaccgccct
      961 gtacgacggc accgcctcgg acggcgagca ggcgctctac atgagcatgc cctgccccta
     1021 actcgaggca tgcggtaccc acgtgttatg cttccggctc ctatgttgtg tggaattgtg
     1081 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     1141 aaccctcact aaagggcccc aattggctag cccgcgggcg gccgctggga acaaaagctg
     1201 gagctcatag cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc
     1261 cgcgcatcgc cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc
     1321 ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc
     1381 ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt
     1441 gaaaattttt tttttgattt ttttctcttt cgatgacctc ccattgatat ttaagttaat
     1501 aaacggtctt caatttctca agtttcagtt tcatttttct tgttctatta caactttttt
     1561 tacttcttgc tcattagaaa gaaagcatag caatctaatc taagttttct agaactagtg
     1621 gatcccccgg gttaattaac atggtgagca agggcgagga ggataacatg gccatcatca
     1681 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
     1741 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     1801 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     1861 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     1921 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     1981 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     2041 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     2101 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     2161 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     2221 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     2281 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     2341 tgtacaagta gcttaagtac gtacctaggt cagtactgac aataaaaaga ttcttgtttt
     2401 caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt
     2461 tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc
     2521 gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat
     2581 tcgatactaa cgccgccatc cagtgtcgaa aacgagctcg aattcatcga tgatatcaga
     2641 tccactagtg gcctatgcgg ccgcggatct gccggtctcc ctatagtgag tcgtattaat
     2701 ttcgataagc caggttaacc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2761 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2821 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2881 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2941 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3001 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3061 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3121 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     3181 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3241 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3301 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3361 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3421 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3481 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3541 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3601 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3661 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3721 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3781 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3841 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3901 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3961 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4021 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4081 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4141 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4201 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4261 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4321 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4381 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4441 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4501 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4561 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4621 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4681 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     4741 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     4801 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4861 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     4921 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atggacatat tgtcgttaga
     4981 acgcggctac aattaataca taaccttatg tatcatacac atacgattta ggtgacacta
     5041 ta
//
