LOCUS       sequence-214073-        8493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8493)
  TITLE     yeast expression vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8493)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pTAU-C-DM
FEATURES             Location/Qualifiers
     source          1..8493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      27..615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     516..535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1597..1614
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1722..1937
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             1938..2741
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     protein_bind    2932..2953
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2968..2973,2974..2991,2992..2998)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3006..3022
                     /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     3011..3033
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3030..3046
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3030..3046
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3087..3106
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3334..3350
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3360..3379
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3360..3378
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(3397..3644)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3610..3628
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    3661..3785
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3963..4065
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4066..4749
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     complement(4132..4151)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5069..5374
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     5253..5272
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5803..5927)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6226..6242)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6451..6470)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6570..6592
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6691..7881)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     7510..7529
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(7929..7934,7935..7951,7952..7957))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        8070..8174
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
       61 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
      121 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
      181 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
      241 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      301 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      361 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      421 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
      481 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
      541 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
      601 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
      661 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
      721 gcctttgagt gagcatcgat tagagcatga cggtatattt atagattttg aagtgaggaa
      781 acctagtagt aataattcta ttaatatcaa agtctgaata atagaatgag ttacatacgt
      841 aaacgtgtag tattattatt attattctat ttcctttccg tttttattat ttatatatat
      901 atatatatat atatatattt gggtgtttat atttaggtag ggcataagga tttactgtcg
      961 gcatgaacct acttgaacaa aaaaataaaa atcaaaagaa aaataatgta ggcttttgca
     1021 catcgactat atataaatat atatatacac acttctgtgc agaagaaaag gaataaaacg
     1081 tcaaactaat ttatttcttc ttgcaacgat tccgacttct cctttaaacg tgcaatttcg
     1141 tcatcgcgat gggtcgaagt tgttaatgct aaatactcca agaaaaagaa caacaaaaat
     1201 aagtaaaata atatataaac ctgtataata taaccttgaa gactatattt cttttcatca
     1261 cgtgctataa aaataattat aatttaaatt ttttaatata aatatataaa ttaaaaatag
     1321 aaagtaaaaa aagaaattaa agaaaaaata gtttttgttt tccgaagatg taaaataggt
     1381 tgaaagttag aaattagtat tataatagca aaaaaaattt aaagttagaa attagaattt
     1441 aaggctctac acacgtttac gatgatattg gacgaccgac acgaaaagac agttttaggc
     1501 tttcttttgt cttgctcttg cgttgaaagt acgtaatttc cattcgactc tagcatcgat
     1561 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     1621 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     1681 agatctgagc agattgtact gagagtgcac cataccacct tttcaattca tcattttttt
     1741 tttattcttt tttttgattt cggtttcctt gaaatttttt tgattcggta atctccgaac
     1801 agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc atatgtagtg
     1861 ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca aaaacctgca
     1921 ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct actcatccta
     1981 gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt
     2041 cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta ggtcccaaaa
     2101 tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag ggcacagtta
     2161 agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaagac agaaaatttg
     2221 ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga atagcagaat
     2281 gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc ggtttgaagc
     2341 aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca gaattgtcat
     2401 gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt gcgaagagcg
     2461 acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga gatgaaggtt
     2521 acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagac gcattgggtc
     2581 aacagtatag aaccgtggat gatgtggtct ctacaggatc tgacattatt attgttggaa
     2641 gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac agaaaagcag
     2701 gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta ttataagtaa
     2761 atgcatgtat actaaactca caaattagag cttcaattta attatatcag ttattaccct
     2821 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gaaagatctg
     2881 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     2941 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     3001 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     3061 gcttgggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3121 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3181 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     3241 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     3301 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt
     3361 aatacgactc actatagggc gaattgggta ccggccgcaa attaaagcct tcgagcgtcc
     3421 caaaaccttc tcaagcaagg ttttcagtat aatgttacat gcgtacacgc gtctgtacag
     3481 aaaaaaaaga aaaatttgaa atataaataa cgttcttaat actaacataa ctataaaaaa
     3541 ataaataggg acctagactt caggttgtct aactccttcc ttttcggtta gagcggatgt
     3601 ggggggaggg cgtgaatgta agcgtgacat aactaattac atgactcgag ttagttactt
     3661 accactttgt acaagaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta
     3721 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca
     3781 ctatgaatca actacttaga tggtattagt gacctgtagt cgactaagtt ggcagcatca
     3841 cccgacgcac tttgcgccga ataaatacct gtgacggaag atcacttcgc agaataaata
     3901 aatcctggtg tccctgttga taccgggaag ccctgggcca acttttggcg aaaatgagac
     3961 gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
     4021 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     4081 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     4141 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     4201 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     4261 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
     4321 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     4381 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     4441 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     4501 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     4561 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     4621 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     4681 aatgaattac aacagtactg cgatgagtgg cagggggggc gtaaacgccg cgtggatccg
     4741 gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa
     4801 tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat
     4861 tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc
     4921 tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg
     4981 aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct
     5041 tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga
     5101 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg
     5161 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
     5221 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     5281 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
     5341 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca
     5401 gtctgcaggt cgatacagta gaaattacag aaactttatc acgtttagta agtatagagg
     5461 ctgaaaatcc agatgaagcc gaacgacttg taagagaaaa gtataagagt tgtgaaattg
     5521 ttcttgatgc agatgatttt caggactatg acactagcgt atatgaatag gtagatgttt
     5581 ttattttgtc acacaaaaaa gaggctcgca cctctttttc ttatttcttt ttatgattta
     5641 atacggcatt gaggacaata gcgagtaggc tggatacgac gattccgttt gagaagaaca
     5701 tttggaaggc tgtcggtcga ctaagttggc agcatcaccc gaagaacatt tggaaggctg
     5761 tcggtcgact acaggtcact aataccatct aagtagttga ttcatagtga ctggatatgt
     5821 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
     5881 tttatatcat tttacgtttc tcgttcagct tttttgtaca aacttgtttc tcgaggtcga
     5941 ggcggccgcg atatcggatc cgtcgacccc attttatgtg atgattgatt gattgtacag
     6001 tttgtttttc ttaatatcta tttcgatgac ttctatatga tattgcacta acaagaagat
     6061 attataatgc aattgataca agacaaggag ttatttgctt ctcttttata tgattctgac
     6121 aatccatatt gcgttggtag tcttttttgc tggaacggtt cagcggaaaa gacgcatcgc
     6181 tctttttgct tctagaagaa atgccagcaa aaggagctcg aattcactgg ccgtcgtttt
     6241 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     6301 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     6361 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     6421 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     6481 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     6541 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     6601 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     6661 tgtcatgata ataatggttt cttagacggg tcaggtcgag gtggcccggc tccatgcacc
     6721 gcgacgcaac gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc
     6781 gttccatgtg ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga
     6841 agttaggctg gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac
     6901 ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat
     6961 cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc
     7021 gtcggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
     7081 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
     7141 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
     7201 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
     7261 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgacgctc
     7321 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     7381 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     7441 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     7501 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     7561 gccggccacg atgcgtccgg cgtagaggat ccacaggacg ggtgtggtcg ccatgatcgc
     7621 gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc
     7681 ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag
     7741 cacgccatag tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag
     7801 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
     7861 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
     7921 actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa
     7981 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     8041 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     8101 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     8161 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc cggtatcatt
     8221 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     8281 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     8341 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     8401 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     8461 taacgtgagt tttcgttcca ctgagcgtca gac
//
