LOCUS       sequence-211499-        8891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses yeast NAB3 using the endogenous promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8891)
  AUTHORS   Loya TJ, O'Rourke TW, Reines D
  TITLE     A genetic screen for terminator function in yeast identifies a role 
            for a new functional domain in termination factor Nab3.
  JOURNAL   Nucleic Acids Res. 2012 Aug;40(15):7476-91. doi: 10.1093/nar/gks377.
            Epub 2012 May 7.
  PUBMED    22564898
REFERENCE   2  (bases 1 to 8891)
  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"; doi: 
            "10.1093/nar/gks377"; journalName: "Nucleic Acids Res"; date: 
            "2012-08"; volume: "40"; issue: "15"; pages: "7476-91"
COMMENT     Sequence Label: pRS315Nab3-11
FEATURES             Location/Qualifiers
     source          1..8891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..136
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     128..145
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     129..145
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     152..171
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        152..170
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     203..219
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(3183..3201)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3222..3238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3222..3238)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3235..3257)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3246..3262
                     /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(3270..3276,3277..3294,3295..3300))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3315..3336
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3453..3470)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3624..4212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3704..3723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4383..5174,5175..5243))
                     /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"
     primer_bind     5006..5025
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5244..5348)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5385..5888
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5930..5948
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5986..6008)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6108..6127
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(6621..7715)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(7716..8123)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(8424..8879)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8561..8582)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8773..8792
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
       61 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
      121 acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattgga
      181 gctccaccgc ggtggcggcc gctctagaac tagtggatcc agtgtaaccc tgaattgttg
      241 aagagaaaat gaagcggaag aagcaaagga caaaaaacaa ttcatttgat tttgccactt
      301 ctaaatgaag gtctaataaa agctatcttg agcatcttta ttagattctg cacagcaaca
      361 agcgattttc tttggtcaaa atataataat tgacttacgt ttttcccgga ctgtcctttc
      421 ataatataat aaccatctgc aagccatgtc agatgaaaac cataacagtg atgttcaaga
      481 tattccttca cctgaactat ccgtcgatag taactctaac gagaatgaat tgatgaataa
      541 ctcaagcgca gacgatggaa tcgaatttga cgccccagag gaagaaagag aagccgaaag
      601 ggaggaggaa aatgaagaac aacacgaact ggaagatgtg aacgatgaag aggaggaaga
      661 taaggaggaa aaaggagagg aaaacgggga agtaataaac acagaagaag aagaagaaga
      721 agaacatcaa caaaaaggcg gaaatgatga tgacgatgat gataatgaag aggaagaaga
      781 ggaagaagag gatgatgacg atgatgatga tgacgacgat gatgatgaag aagaagaaga
      841 agaagaagaa gaagaaggca acgacaacag ttcggtaggc tcagatagtg ccgctgaaga
      901 cggtgaggat gaggaagaca aaaaggataa aaccaaagat aaagaggtcg aacttcgccg
      961 tgaaacattg gaaaaagaac aaaaggacgt agatgaagct ataaaaaaaa taactcgtga
     1021 agaaaatgat aatactcact ttccaactaa tatggaaaat gttaattacg atcttttaca
     1081 aaagcaagtc aagtacatta tggacagtaa catgctaaat ttgcctcagt ttcaacattt
     1141 acctcaagaa gaaaagatgt ctgcgatttt agcaatgtta aattcaaatt ctgacacagc
     1201 tctttccgta cctcctcatg atagtactat ctcaacaaca gctagcgcct cagccacaag
     1261 cggcgcaaga agcaatgatc aaagaaaacc tccattgtca gatgcccaaa gacgtatgag
     1321 atttcctagg gcggatttat ctaagccgat taccgaagaa gaacacgacc gttatgcagc
     1381 ctatttgcac ggtgaaaata aaatcaccga gatgcacaat attcctccga agtcaagatt
     1441 attcattggt aatttgccgc taaagaacgt ttctaaggag gatttattta ggattttctc
     1501 tccatacggt catatcatgc aaatcaatat caaaaatgcc tttggattca ttcagcttga
     1561 caacactcaa agcgttagag atgcaattga atgcgagtct caagaaatga actttggcaa
     1621 aaagttgatc ctggaagttt ctagctcgaa tgctcgtcct caatttgatc atggtgatca
     1681 cggtacaaac agtagttcta cttttatttc ttccgcaaaa cgaccatttc aaactgaatc
     1741 tggtgacatg tacaatgacg acaatggtgc tggctacaag aaatccagaa gacacaccgt
     1801 ttcttgcaac attttcgtta aaagaaccgc agatcgtacg tatgccattg aggttttcaa
     1861 caggtttagg gacgggactg gtttggaaac tgatatgatt ttcttgaaac caagaatgga
     1921 actgggaaag cttatcaatg atgccgcata taatggggtg tggggcgttg ttttagttaa
     1981 taaaacacac aatgtagatg ttcaaacttt ctacaaaggc tcacaaggtg aaacgaaatt
     2041 tgatgaatat attagcatat ccgctgatga cgcagttgcc atttttaata acatcaaaaa
     2101 caacagaaat aattctcgtc ctactgatta ccgtgctatg agccatcagc aaaacatata
     2161 tggcgctcct cctcttcctg ttccaaacgg cccagctgtc ggacctcctc ctcaaacaaa
     2221 ctattaccag ggttacagta tgcctcctcc acaacaacaa cagcaacagc catatggtaa
     2281 ttatgggatg ccaccaccat cccatgacca aggatatggt tctcaacctc caattccaat
     2341 gaatcagagc tacggtcgct accagacttc tattccacca ccacctccac aacaacaaat
     2401 tcctcaaggg tatggtcgtt atcaggctgg tccgcctcct caaccacctt ctcaaactcc
     2461 aatggaccag caacaactat tatctgccat tcaaaacctt ccacctaacg ttgtatcgaa
     2521 tttgctttca atggcccaac aacagcaaca acaacctcat gctcagcagc aattggttgg
     2581 tttaatacaa tcaatgcaag gccaggctcc tcaacaacag caacaacagt tgggtggata
     2641 ttcctctatg aactcatcct ctccccctcc tatgagtacc aattacaatg gtcaaaatat
     2701 atctgcaaaa ccctctgccc caccaatgtc acaccaacct ccgccacctc aacaacaaca
     2761 acaacaacaa caacagcaac agcaacagca acagcaacct gctggcaata atgttcaaag
     2821 tctattagat agtttagcaa aactacaaaa atagactccc ttttttcaat cttttccatt
     2881 tcttgtacat tatacttgaa ttcttttaat tggacgaaaa atttttaagg aaagtagatt
     2941 gatatggttt gcataaattc gacatagcgg tttatgcttt aatcgttttc ccttcccaaa
     3001 ttggactagg acttaattct tactattgct gcatctcttt cagaacgaaa aactctttac
     3061 cctgttataa atttctttta tttcttcctt atcaaatgct atttggtcta tttatttcac
     3121 ttacacgcac ctcttcattt gtttcctctg ctcgaggggg ggcccggtac cagcttttgt
     3181 tccctttagt gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg
     3241 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3301 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3361 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3421 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3481 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3541 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3601 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3661 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3721 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3781 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3841 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3901 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3961 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4021 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4081 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4141 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4201 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4261 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4321 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4381 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4441 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4501 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4561 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4621 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4681 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4741 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4801 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4861 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4921 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4981 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5041 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5101 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5161 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5221 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5281 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5341 gttccgcgca catttccccg aaaagtgcca cctgggtcct tttcatcacg tgctataaaa
     5401 ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa
     5461 gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc
     5521 caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt
     5581 catcttgtct gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg
     5641 ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt
     5701 acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc
     5761 ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac
     5821 agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacaggc
     5881 aagcgatccg tcctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5941 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     6001 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     6061 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     6121 gattgtactg agagtgcacc atatcgacta cgtcgttaag gccgtttctg acagagtaaa
     6181 attcttgagg gaactttcac cattatggga aatggttcaa gaaggtattg acttaaactc
     6241 catcaaatgg tcaggtcatt gagtgttttt tatttgttgt attttttttt ttttagagaa
     6301 aatcctccaa tatataaatt aggaatcata gtttcatgat tttctgttac acctaacttt
     6361 ttgtgtggtg ccctcctcct tgtcaatatt aatgttaaag tgcaattctt tttccttatc
     6421 acgttgagcc attagtatca atttgcttac ctgtattcct ttacatcctc ctttttctcc
     6481 ttcttgataa atgtatgtag attgcgtata tagtttcgtc taccctatga acatattcca
     6541 ttttgtaatt tcgtgtcgtt tctattatga atttcattta taaagtttat gtacaaatat
     6601 cataaaaaaa gagaatcttt ttaagcaagg attttcttaa cttcttcggc gacagcatca
     6661 ccgacttcgg tggtactgtt ggaaccacct aaatcaccag ttctgatacc tgcatccaaa
     6721 acctttttaa ctgcatcttc aatggcctta ccttcttcag gcaagttcaa tgacaatttc
     6781 aacatcattg cagcagacaa gatagtggcg atagggttga ccttattctt tggcaaatct
     6841 ggagcagaac cgtggcatgg ttcgtacaaa ccaaatgcgg tgttcttgtc tggcaaagag
     6901 gccaaggacg cagatggcaa caaacccaag gaacctggga taacggaggc ttcatcggag
     6961 atgatatcac caaacatgtt gctggtgatt ataataccat ttaggtgggt tgggttctta
     7021 actaggatca tggcggcaga atcaatcaat tgatgttgaa ccttcaatgt agggaattcg
     7081 ttcttgatgg tttcctccac agtttttctc cataatcttg aagaggccaa aacattagct
     7141 ttatccaagg accaaatagg caatggtggc tcatgttgta gggccatgaa agcggccatt
     7201 cttgtgattc tttgcacttc tggaacggtg tattgttcac tatcccaagc gacaccatca
     7261 ccatcgtctt cctttctctt accaaagtaa atacctccca ctaattctct gacaacaacg
     7321 aagtcagtac ctttagcaaa ttgtggcttg attggagata agtctaaaag agagtcggat
     7381 gcaaagttac atggtcttaa gttggcgtac aattgaagtt ctttacggat ttttagtaaa
     7441 ccttgttcag gtctaacact accggtaccc catttaggac cacccacagc acctaacaaa
     7501 acggcatcag ccttcttgga ggcttccagc gcctcatctg gaagtggaac acctgtagca
     7561 tcgatagcag caccaccaat taaatgattt tcgaaatcga acttgacatt ggaacgaaca
     7621 tcagaaatag ctttaagaac cttaatggct tcggctgtga tttcttgacc aacgtggtca
     7681 cctggcaaaa cgacgatctt cttaggggca gacattagaa tggtatatcc ttgaaatata
     7741 tatatatata ttgctgaaat gtaaaaggta agaaaagtta gaaagtaaga cgattgctaa
     7801 ccacctattg gaaaaaacaa taggtcctta aataatattg tcaacttcaa gtattgtgat
     7861 gcaagcattt agtcatgaac gcttctctat tctatatgaa aagccggttc cggcgctctc
     7921 acctttcctt tttctcccaa tttttcagtt gaaaaaggta tatgcgtcag gcgacctctg
     7981 aaattaacaa aaaatttcca gtcatcgaat ttgattctgt gcgatagcgc ccctgtgtgt
     8041 tctcgttatg ttgaggaaaa aaataatggt tgctaagaga ttcgaactct tgcatcttac
     8101 gatacctgag tattcccaca gttaactgcg gtcaagatat ttcttgaatc aggcgcctta
     8161 gaccgctcgg ccaaacaacc aattacttgt tgagaaatag agtataatta tcctataaat
     8221 ataacgtttt tgaacacaca tgaacaagga agtacaggac aattgatttt gaagagaatg
     8281 tggattttga tgtaattgtt gggattccat ttttaataag gcaataatat taggtatgta
     8341 gatatactag aagttctcct cgaccggtcg atatgcggtg tgaaataccg cacagatgcg
     8401 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     8461 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     8521 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     8581 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     8641 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     8701 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     8761 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     8821 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     8881 cattcgccat t
//
