LOCUS       sequence-213355-        7767 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 7767)
  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 7767)
  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: pRS316Nab3
FEATURES             Location/Qualifiers
     source          1..7767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(2557..2575)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2596..2612)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2596..2612)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2609..2631)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2620..2636
                     /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(2644..2650,2651..2668,2669..2674))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2689..2710
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2827..2844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2998..3586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3078..3097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3757..4548,4549..4617))
                     /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     4380..4399
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4618..4722)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4759..5262
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5303..5321
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5359..5381)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5481..5500
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5524..5739
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5740..6543
                     /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"
     rep_origin      complement(6674..7129)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6811..6832)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7023..7042
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7255..7277
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7269..7286
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7270..7286
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7293..7312
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7293..7311
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7344..7360
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7344..7360
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 aactggaaga tgtgaacgat gaagaggagg aagataagga ggaaaaagga gaggaaaacg
       61 gggaagtaat aaacacagaa gaagaagaag aagaagaaca tcaacaaaaa ggcggaaatg
      121 atgatgacga tgatgataat gaagaggaag aagaggaaga agaggatgat gacgatgatg
      181 atgatgacga cgatgatgat gaagaagaag aagaagaaga agaagaagaa ggcaacgaca
      241 acagttcggt aggctcagat agtgccgctg aagacggtga ggatgaggaa gacaaaaagg
      301 ataaaaccaa agataaagag gtcgaacttc gccgtgaaac attggaaaaa gaacaaaagg
      361 acgtagatga agctataaaa aaaataactc gtgaagaaaa tgataatact cactttccaa
      421 ctaatatgga aaatgttaat tacgatcttt tacaaaagca agtcaagtac attatggaca
      481 gtaacatgct aaatttgcct cagtttcaac atttacctca agaagaaaag atgtctgcga
      541 ttttagcaat gttaaattca aattctgaca cagctctttc cgtacctcct catgatagta
      601 ctatctcaac aacagctagc gcctcagcca caagcggcgc aagaagcaat gatcaaagaa
      661 aacctccatt gtcagatgcc caaagacgta tgagatttcc tagggcggat ttatctaagc
      721 cgattaccga agaagaacac gaccgttatg cagcctattt gcacggtgaa aataaaatca
      781 ccgagatgca caatattcct ccgaagtcaa gattattcat tggtaatttg ccgctaaaga
      841 acgtttctaa ggaggattta tttaggattt tctctccata cggtcatatc atgcaaatca
      901 atatcaaaaa tgcctttgga ttcattcagt ttgacaaccc tcaaagcgtt agagatgcaa
      961 ttgaatgcga gtctcaagaa atgaactttg gcaaaaagtt gatcctggaa gtttctagct
     1021 cgaatgctcg tcctcaattt gatcatggtg atcacggtac aaacagtagt tctactttta
     1081 tttcttccgc aaaacgacca tttcaaactg aatctggtga catgtacaat gacgacaatg
     1141 gtgctggcta caagaaatcc agaagacaca ccgtttcttg caacattttc gttaaaagaa
     1201 ccgcagatcg tacgtatgcc attgaggttt tcaacaggtt tagggacggg actggtttgg
     1261 aaactgatat gattttcttg aaaccaagaa tggaactggg aaagcttatc aatgatgccg
     1321 catataatgg ggtgtggggc gttgttttag ttaataaaac acacaatgta gatgttcaaa
     1381 ctttctacaa aggctcacaa ggtgaaacga aatttgatga atatattagc atatccgctg
     1441 atgacgcagt tgccattttt aataacatca aaaacaacag aaataattct cgtcctactg
     1501 attaccgtgc tatgagccat cagcaaaaca tatatggcgc tcctcctctt cctgttccaa
     1561 acggcccagc tgtcggacct cctcctcaaa caaactatta ccagggttac agtatgcctc
     1621 ctccacaaca acaacagcaa cagccatatg gtaattatgg gatgccacca ccatcccatg
     1681 accaaggata tggttctcaa cctccaattc caatgaatca gagctacggt cgctaccaga
     1741 cttctattcc accaccacct ccacaacaac aaattcctca agggtatggt cgttatcagg
     1801 ctggtccgcc tcctcaacca ccttctcaaa ctccaatgga ccagcaacaa ctattatctg
     1861 ccattcaaaa ccttccacct aacgttgtat cgaatttgct ttcaatggcc caacaacagc
     1921 aacaacaacc tcatgctcag cagcaattgg ttggtttaat acaatcaatg caaggccagg
     1981 ctcctcaaca acagcaacaa cagttgggtg gatattcctc tatgaactca tcctctcccc
     2041 ctcctatgag taccaattac aatggtcaaa atatatctgc aaaaccctct gccccaccaa
     2101 tgtcacacca acctccgcca cctcaacaac aacaacaaca acaacaacag caacagcaac
     2161 agcaacagca acctgctggc aataatgttc aaagtctatt agatagttta gcaaaactac
     2221 aaaaatagac tccctttttt caatcttttc catttcttgt acattatact tgaattcttt
     2281 taattggacg aaaaattttt aaggaaagta gattgatatg gtttgcataa attcgacata
     2341 gcggtttatg ctttaatcgt tttcccttcc caaattggac taggacttaa ttcttactat
     2401 tgctgcatct ctttcagaac gaaaaactct ttaccctgtt ataaatttct tttatttctt
     2461 ccttatcaaa tgctatttgg tctatttatt tcacttacac gcacctcttc atttgtttcc
     2521 tctgctcgag ggggggcccg gtaccagctt ttgttccctt tagtgagggt taatttcgag
     2581 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2641 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2701 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2761 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2821 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2881 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2941 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3001 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3061 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3121 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3181 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3241 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3301 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3361 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3421 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3481 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3541 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3601 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3661 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3721 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3781 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3841 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3901 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3961 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4021 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4081 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4141 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4201 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4261 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4321 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4381 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4441 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4501 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4561 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4621 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4681 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4741 gccacctggg tccttttcat cacgtgctat aaaaataatt ataatttaaa ttttttaata
     4801 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
     4861 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
     4921 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac
     4981 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
     5041 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
     5101 ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt tactttctaa
     5161 aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc
     5221 attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtctaag aaaccattat
     5281 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     5341 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     5401 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5461 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accataccac
     5521 cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc ttgaaatttt
     5581 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
     5641 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
     5701 ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc tacatataag
     5761 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
     5821 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
     5881 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
     5941 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
     6001 ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
     6061 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
     6121 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
     6181 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
     6241 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
     6301 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
     6361 gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt ctctacagga
     6421 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
     6481 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
     6541 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
     6601 taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6661 ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     6721 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     6781 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     6841 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     6901 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     6961 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     7021 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     7081 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat
     7141 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     7201 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     7261 cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg
     7321 agctccaccg cggtggcggc cgctctagaa ctagtggatc cagtgtaacc ctgaattgtt
     7381 gaagagaaaa tgaagcggaa gaagcaaagg acaaaaaaca attcatttga ttttgccact
     7441 tctaaatgaa ggtctaataa aagctatctt gagcatcttt attagattct gcacagcaac
     7501 aagcgatttt ctttggtcaa aatataataa ttgacttacg tttttcccgg actgtccttt
     7561 cataatataa taaccatctg caagccatgt cagatgaaaa ccataacagt gatgttcaag
     7621 atattccttc acctgaacta tccgtcgata gtaactctaa cgagaatgaa ttgatgaata
     7681 actcaagcgc agacgatgga atcgaatttg acgccccaga ggaagaaaga gaagccgaaa
     7741 gggaggagga aaatgaagaa caacacg
//
