LOCUS       sequence-213347-        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: pRS316Nab3-11
FEATURES             Location/Qualifiers
     source          1..7767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..59
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     51..68
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     52..68
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     75..94
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        75..93
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     126..142
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     126..142
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(3106..3124)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3145..3161)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3145..3161)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3158..3180)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3169..3185
                     /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(3193..3199,3200..3217,3218..3223))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3238..3259
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3376..3393)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3547..4135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3627..3646)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4306..5097,5098..5166))
                     /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     4929..4948
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5167..5271)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5308..5811
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5852..5870
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5908..5930)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6030..6049
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6073..6288
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             6289..7092
                     /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(7223..7678)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7360..7381)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7572..7591
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
       61 cggccagtga attgtaatac gactcactat agggcgaatt ggagctccac cgcggtggcg
      121 gccgctctag aactagtgga tccagtgtaa ccctgaattg ttgaagagaa aatgaagcgg
      181 aagaagcaaa ggacaaaaaa caattcattt gattttgcca cttctaaatg aaggtctaat
      241 aaaagctatc ttgagcatct ttattagatt ctgcacagca acaagcgatt ttctttggtc
      301 aaaatataat aattgactta cgtttttccc ggactgtcct ttcataatat aataaccatc
      361 tgcaagccat gtcagatgaa aaccataaca gtgatgttca agatattcct tcacctgaac
      421 tatccgtcga tagtaactct aacgagaatg aattgatgaa taactcaagc gcagacgatg
      481 gaatcgaatt tgacgcccca gaggaagaaa gagaagccga aagggaggag gaaaatgaag
      541 aacaacacga actggaagat gtgaacgatg aagaggagga agataaggag gaaaaaggag
      601 aggaaaacgg ggaagtaata aacacagaag aagaagaaga agaagaacat caacaaaaag
      661 gcggaaatga tgatgacgat gatgataatg aagaggaaga agaggaagaa gaggatgatg
      721 acgatgatga tgatgacgac gatgatgatg aagaagaaga agaagaagaa gaagaagaag
      781 gcaacgacaa cagttcggta ggctcagata gtgccgctga agacggtgag gatgaggaag
      841 acaaaaagga taaaaccaaa gataaagagg tcgaacttcg ccgtgaaaca ttggaaaaag
      901 aacaaaagga cgtagatgaa gctataaaaa aaataactcg tgaagaaaat gataatactc
      961 actttccaac taatatggaa aatgttaatt acgatctttt acaaaagcaa gtcaagtaca
     1021 ttatggacag taacatgcta aatttgcctc agtttcaaca tttacctcaa gaagaaaaga
     1081 tgtctgcgat tttagcaatg ttaaattcaa attctgacac agctctttcc gtacctcctc
     1141 atgatagtac tatctcaaca acagctagcg cctcagccac aagcggcgca agaagcaatg
     1201 atcaaagaaa acctccattg tcagatgccc aaagacgtat gagatttcct agggcggatt
     1261 tatctaagcc gattaccgaa gaagaacacg accgttatgc agcctatttg cacggtgaaa
     1321 ataaaatcac cgagatgcac aatattcctc cgaagtcaag attattcatt ggtaatttgc
     1381 cgctaaagaa cgtttctaag gaggatttat ttaggatttt ctctccatac ggtcatatca
     1441 tgcaaatcaa tatcaaaaat gcctttggat tcattcagct tgacaacact caaagcgtta
     1501 gagatgcaat tgaatgcgag tctcaagaaa tgaactttgg caaaaagttg atcctggaag
     1561 tttctagctc gaatgctcgt cctcaatttg atcatggtga tcacggtaca aacagtagtt
     1621 ctacttttat ttcttccgca aaacgaccat ttcaaactga atctggtgac atgtacaatg
     1681 acgacaatgg tgctggctac aagaaatcca gaagacacac cgtttcttgc aacattttcg
     1741 ttaaaagaac cgcagatcgt acgtatgcca ttgaggtttt caacaggttt agggacggga
     1801 ctggtttgga aactgatatg attttcttga aaccaagaat ggaactggga aagcttatca
     1861 atgatgccgc atataatggg gtgtggggcg ttgttttagt taataaaaca cacaatgtag
     1921 atgttcaaac tttctacaaa ggctcacaag gtgaaacgaa atttgatgaa tatattagca
     1981 tatccgctga tgacgcagtt gccattttta ataacatcaa aaacaacaga aataattctc
     2041 gtcctactga ttaccgtgct atgagccatc agcaaaacat atatggcgct cctcctcttc
     2101 ctgttccaaa cggcccagct gtcggacctc ctcctcaaac aaactattac cagggttaca
     2161 gtatgcctcc tccacaacaa caacagcaac agccatatgg taattatggg atgccaccac
     2221 catcccatga ccaaggatat ggttctcaac ctccaattcc aatgaatcag agctacggtc
     2281 gctaccagac ttctattcca ccaccacctc cacaacaaca aattcctcaa gggtatggtc
     2341 gttatcaggc tggtccgcct cctcaaccac cttctcaaac tccaatggac cagcaacaac
     2401 tattatctgc cattcaaaac cttccaccta acgttgtatc gaatttgctt tcaatggccc
     2461 aacaacagca acaacaacct catgctcagc agcaattggt tggtttaata caatcaatgc
     2521 aaggccaggc tcctcaacaa cagcaacaac agttgggtgg atattcctct atgaactcat
     2581 cctctccccc tcctatgagt accaattaca atggtcaaaa tatatctgca aaaccctctg
     2641 ccccaccaat gtcacaccaa cctccgccac ctcaacaaca acaacaacaa caacaacagc
     2701 aacagcaaca gcaacagcaa cctgctggca ataatgttca aagtctatta gatagtttag
     2761 caaaactaca aaaatagact cccttttttc aatcttttcc atttcttgta cattatactt
     2821 gaattctttt aattggacga aaaattttta aggaaagtag attgatatgg tttgcataaa
     2881 ttcgacatag cggtttatgc tttaatcgtt ttcccttccc aaattggact aggacttaat
     2941 tcttactatt gctgcatctc tttcagaacg aaaaactctt taccctgtta taaatttctt
     3001 ttatttcttc cttatcaaat gctatttggt ctatttattt cacttacacg cacctcttca
     3061 tttgtttcct ctgctcgagg gggggcccgg taccagcttt tgttcccttt agtgagggtt
     3121 aatttcgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3181 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3241 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3301 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3361 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3421 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3481 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3541 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3601 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3661 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3721 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3781 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3841 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3901 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3961 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4021 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4081 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4141 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4201 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4261 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4321 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4381 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4441 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4501 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4561 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4621 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4681 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4741 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4801 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4861 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4921 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4981 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5041 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5101 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5161 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5221 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5281 ccgaaaagtg ccacctgggt ccttttcatc acgtgctata aaaataatta taatttaaat
     5341 tttttaatat aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat
     5401 agtttttgtt ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt
     5461 tatcttgctc ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga
     5521 agaccacaca cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc
     5581 tatttaatct gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat
     5641 tttttaaacc tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt
     5701 actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat
     5761 tattccatca ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtctaaga
     5821 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     5881 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     5941 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     6001 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6061 ccataccacc ttttcaattc atcatttttt ttttattctt ttttttgatt tcggtttcct
     6121 tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga
     6181 cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc
     6241 ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa gataaatcat gtcgaaagct
     6301 acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc
     6361 atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta
     6421 ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc
     6481 ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac
     6541 aattttttac tcttcgaaga cagaaaattt gctgacattg gtaatacagt caaattgcag
     6601 tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg
     6661 gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct
     6721 agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat
     6781 actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct
     6841 caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg
     6901 ggtttagatg acaagggaga cgcattgggt caacagtata gaaccgtgga tgatgtggtc
     6961 tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct
     7021 aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc
     7081 cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga
     7141 gcttcaattt aattatatca gttattaccc tatgcggtgt gaaataccgc acagatgcgt
     7201 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta
     7261 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     7321 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     7381 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     7441 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     7501 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     7561 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     7621 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     7681 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     7741 tacgccagct ggcgaaaggg ggatgtg
//
