LOCUS       sequence-484590-        6654 bp    DNA     circular SYN 25-AUG-2025
DEFINITION  Backbone for PYR1 sensor library assembly from oligopools; stop 
            codon removed; PYR1 sensor library assembly from oligopools; .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6654)
  TITLE     Cutler lab 2025
REFERENCE   2  (bases 1 to 6654)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pBD_PYR1_156-F1
FEATURES             Location/Qualifiers
     source          1..6654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        457..558
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             559..1218
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(625..644)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1499..1517
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1555..1577)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1677..1696
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      1713..2877
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        2882..3283
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             3311..3751
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     primer_bind     complement(3957..3976)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3958..3976)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3998..4185
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4053..4077)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(4238..4912)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(4913..5014)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5028..5044)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5028..5044)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5041..5063)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5052..5068
                     /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)."
     promoter        complement(join(5076..5082,5083..5100,5101..5106))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5121..5142
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        complement(5214..5318)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5345..5800
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5432..5451)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5642..5663
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5895..5912)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6066..6654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6146..6165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 gcctgtgacg gaagatcact tcgcagaata aataaatcct ggtgtccctg ttgataccgg
      421 gaagccctgg gccaactttt ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa
      481 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagtttc gagattttca
      541 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
      601 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
      661 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
      721 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt
      781 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
      841 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
      901 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
      961 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     1021 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     1081 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     1141 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     1201 tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggttgc
     1261 tacgcctgaa taagtgataa taagcggatg aatggcagaa attcgaaagc aaattcgacc
     1321 cggtcgtcgg ttcagggcag ggtcgttaaa tagccgctta tgtctattgc tggtttaccg
     1381 gtttattgac taccggaagc agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg
     1441 ctcaggctct ccccgtggag gctaagaaac cattattatc atgacattaa cctataaaaa
     1501 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     1561 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     1621 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     1681 atcagagcag attgtactga gagtgcacca taacgcattt aagcataaac acgcactatg
     1741 ccgttcttct catgtatata tatatacagg caacacgcag atataggtgc gacgtgaaca
     1801 gtgagctgta tgtgcgcagc tcgcgttgca ttttcggaag cgctcgtttt cggaaacgct
     1861 ttgaagttcc tattccgaag ttcctattct ctagctagaa agtataggaa cttcagagcg
     1921 cttttgaaaa ccaaaagcgc tctgaagacg cactttcaaa aaaccaaaaa cgcaccggac
     1981 tgtaacgagc tactaaaata ttgcgaatac cgcttccaca aacattgctc aaaagtatct
     2041 ctttgctata tatctctgtg ctatatccct atataaccta cccatccacc tttcgctcct
     2101 tgaacttgca tctaaactcg acctctacat tttttatgtt tatctctagt attactcttt
     2161 agacaaaaaa attgtagtaa gaactattca tagagtgaat cgaaaacaat acgaaaatgt
     2221 aaacatttcc tatacgtagt atatagagac aaaatagaag aaaccgttca taattttctg
     2281 accaatgaag aatcatcaac gctatcactt tctgttcaca aagtatgcgc aatccacatc
     2341 ggtatagaat ataatcgggg atgcctttat cttgaaaaaa tgcacccgca gcttcgctag
     2401 taatcagtaa acgcgggaag tggagtcagg ctttttttat ggaagagaaa atagacacca
     2461 aagtagcctt cttctaacct taacggacct acagtgcaaa aagttatcaa gagactgcat
     2521 tatagagcgc acaaaggaga aaaaaagtaa tctaagatgc tttgttagaa aaatagcgct
     2581 ctcgggatgc atttttgtag aacaaaaaag aagtatagat tctttgttgg taaaatagcg
     2641 ctctcgcgtt gcatttctgt tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta
     2701 gcgctctcgc gttgcatttt tgttttacaa aaatgaagca cagattcttc gttggtaaaa
     2761 tagcgctttc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa
     2821 aattagcgct ctcgcgttgc atttttgttc tacaaaatga agcacagatg cttcgttgct
     2881 tgcatgcaac ttcttttctt tttttttctt ttctctctcc cccgttgttg tctcaccata
     2941 tccgcaatga caaaaaaatg atggaagaca ctaaaggaaa aaattaacga caaagacagc
     3001 accaacagat gtcgttgttc cagagctgat gaggggtatc tcgaagcaca cgaaactttt
     3061 tccttccttc attcacgcac actactctct aatgagcaac ggtatacggc cttccttcca
     3121 gttacttgaa tttgaaataa aaaaaagttt gctgtcttgc tatcaagtat aaatagacct
     3181 gcaattatta atcttttgtt tcctcgtcat tgttctcgtt ccctttcttc cttgtttctt
     3241 tttctgcaca atatttcaag ctataccaag catacaatca actccaagct tgaagcaagc
     3301 ctcctgaaag atgaagctac tgtcttctat cgaacaagca tgcgatattt gccgacttaa
     3361 aaagctcaag tgctccaaag aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga
     3421 gtgtcgctac tctcccaaaa ccaaaaggtc cccgctgact agggcacatc tgacagaagt
     3481 ggaatcaagg ctagaaagac tggaacagct atttctactg atttttcctc gagaagacct
     3541 tgacatgatt ttgaaaatgg attctttaca ggatataaaa gcattgttaa caggattatt
     3601 tgtacaagat aatgtgaata aagatgccgt cacagataga ttggcttcag tggagactga
     3661 tatgcctcta acattgagac agcatagaat aagtgcgaca tcatcatcgg aagagagtag
     3721 taacaaaggt caaagacagt tgactgtatc gccggaattc atgccttcgg agttaacacc
     3781 agaagaacga tcggaactaa aaaactcaat cgccgagttc cacacatacc aactcgatcc
     3841 aggaagctgt tcatcactcc acgcgcaacg aatccacgcg cctccggaac tcgtctggtc
     3901 aatcgtacga cgattcggag acctcgaggt ctcgtgacgt gaggtcgact ctagagccct
     3961 atagtgagtc gtattactgc agccaagcta attccgggcg aatttcttat gatttatgat
     4021 ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa gtgactctta
     4081 ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca ggttgctttc
     4141 tcaggtatag catgaggtcg ctcttattga ccacacctct accggcatgc cggcaagtgc
     4201 acaaacaata cttaaataaa tactactcag taataaccta tttcttagca tttttgacga
     4261 aatttgctat tttgttagag tcttttacac catttgtctc cacacctccg cttacatcaa
     4321 caccaataac gccatttaat ctaagcgcat caccaacatt ttctggcgtc agtccaccag
     4381 ctaacataaa atgtaagctt tcggggctct cttgccttcc aacccagtca gaaatcgagt
     4441 tccaatccaa aagttcacct gtcccacctg cttctgaatc aaacaaggga ataaacgaat
     4501 gaggtttctg tgaagctgca ctgagtagta tgttgcagtc ttttggaaat acgagtcttt
     4561 taataactgg caaaccgagg aactcttggt attcttgcca cgactcatct ccatgcagtt
     4621 ggacgatatc aatgccgtaa tcattgacca gagccaaaac atcctcctta ggttgattac
     4681 gaaacacgcc aaccaagtat ttcggagtgc ctgaactatt tttatatgct tttacaagac
     4741 ttgaaatttt ccttgcaata accgggtcaa ttgttctctt tctattgggc acacatataa
     4801 tacccagcaa gtcagcatcg gaatctagag cacattctgc ggcctctgtg ctctgcaagc
     4861 cgcaaacttt caccaatgga ccagaactac ctgtgaaatt aataacagac atactccaag
     4921 ctgcctttgt gtgcttaatc acgtatactc acgtgctcaa tagtcaccaa tgccctccct
     4981 cttggccctc tccttttctt ttttcgaccg aattaattcg taatcatgtc atagctgttt
     5041 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5101 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5161 cccgctttcc agtcgggaaa cctgtcgtgc cagctggttc tttccgcctc aggactcttc
     5221 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5281 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5341 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5401 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5461 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5521 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5581 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5641 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5701 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5761 tttaacgcga attttaacaa aatattaacg cttacaattt acgcgcctaa attatgcggc
     5821 cgcatataac tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5881 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5941 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6001 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6061 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6241 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6481 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6541 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6601 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaa
//
