LOCUS       sequence-211512-        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
FEATURES             Location/Qualifiers
     source          1..8891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..137
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     129..146
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     130..146
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     153..172
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        153..171
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     204..220
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(3184..3202)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3223..3239)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3223..3239)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3236..3258)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3247..3263
                     /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(3271..3277,3278..3295,3296..3301))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3316..3337
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3454..3471)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3625..4213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3705..3724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4384..5175,5176..5244))
                     /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     5007..5026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5245..5349)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5386..5889
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5931..5949
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5987..6009)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6109..6128
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(6622..7716)
                     /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(7717..8124)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(8425..8880)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8562..8583)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8774..8793
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
       61 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      121 cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg
      181 agctccaccg cggtggcggc cgctctagaa ctagtggatc cagtgtaacc ctgaattgtt
      241 gaagagaaaa tgaagcggaa gaagcaaagg acaaaaaaca attcatttga ttttgccact
      301 tctaaatgaa ggtctaataa aagctatctt gagcatcttt attagattct gcacagcaac
      361 aagcgatttt ctttggtcaa aatataataa ttgacttacg tttttcccgg actgtccttt
      421 cataatataa taaccatctg caagccatgt cagatgaaaa ccataacagt gatgttcaag
      481 atattccttc acctgaacta tccgtcgata gtaactctaa cgagaatgaa ttgatgaata
      541 actcaagcgc agacgatgga atcgaatttg acgccccaga ggaagaaaga gaagccgaaa
      601 gggaggagga aaatgaagaa caacacgaac tggaagatgt gaacgatgaa gaggaggaag
      661 ataaggagga aaaaggagag gaaaacgggg aagtaataaa cacagaagaa gaagaagaag
      721 aagaacatca acaaaaaggc ggaaatgatg atgacgatga tgataatgaa gaggaagaag
      781 aggaagaaga ggatgatgac gatgatgatg atgacgacga tgatgatgaa gaagaagaag
      841 aagaagaaga agaagaaggc aacgacaaca gttcggtagg ctcagatagt gccgctgaag
      901 acggtgagga tgaggaagac aaaaaggata aaaccaaaga taaagaggtc gaacttcgcc
      961 gtgaaacatt ggaaaaagaa caaaaggacg tagatgaagc tataaaaaaa ataactcgtg
     1021 aagaaaatga taatactcac tttccaacta atatggaaaa tgttaattac gatcttttac
     1081 aaaagcaagt caagtacatt atggacagta acatgctaaa tttgcctcag tttcaacatt
     1141 tacctcaaga agaaaagatg tctgcgattt tagcaatgtt aaattcaaat tctgacacag
     1201 ctctttccgt acctcctcat gatagtacta tctcaacaac agctagcgcc tcagccacaa
     1261 gcggcgcaag aagcaatgat caaagaaaac ctccattgtc agatgcccaa agacgtatga
     1321 gatttcctag ggcggattta tctaagccga ttaccgaaga agaacacgac cgttatgcag
     1381 cctatttgca cggtgaaaat aaaatcaccg agatgcacaa tattcctccg aagtcaagat
     1441 tattcattgg taatttgccg ctaaagaacg tttctaagga ggatttattt aggattttct
     1501 ctccatacgg tcatatcatg caaatcaata tcaaaaatgc ctttggattc attcagtttg
     1561 acaaccctca aagcgttaga gatgcaattg aatgcgagtc tcaagaaatg aactttggca
     1621 aaaagttgat cctggaagtt tctagctcga atgctcgtcc tcaatttgat catggtgatc
     1681 acggtacaaa cagtagttct acttttattt cttccgcaaa acgaccattt caaactgaat
     1741 ctggtgacat gtacaatgac gacaatggtg ctggctacaa gaaatccaga agacacaccg
     1801 tttcttgcaa cattttcgtt aaaagaaccg cagatcgtac gtatgccatt gaggttttca
     1861 acaggtttag ggacgggact ggtttggaaa ctgatatgat tttcttgaaa ccaagaatgg
     1921 aactgggaaa gcttatcaat gatgccgcat ataatggggt gtggggcgtt gttttagtta
     1981 ataaaacaca caatgtagat gttcaaactt tctacaaagg ctcacaaggt gaaacgaaat
     2041 ttgatgaata tattagcata tccgctgatg acgcagttgc catttttaat aacatcaaaa
     2101 acaacagaaa taattctcgt cctactgatt accgtgctat gagccatcag caaaacatat
     2161 atggcgctcc tcctcttcct gttccaaacg gcccagctgt cggacctcct cctcaaacaa
     2221 actattacca gggttacagt atgcctcctc cacaacaaca acagcaacag ccatatggta
     2281 attatgggat gccaccacca tcccatgacc aaggatatgg ttctcaacct ccaattccaa
     2341 tgaatcagag ctacggtcgc taccagactt ctattccacc accacctcca caacaacaaa
     2401 ttcctcaagg gtatggtcgt tatcaggctg gtccgcctcc tcaaccacct tctcaaactc
     2461 caatggacca gcaacaacta ttatctgcca ttcaaaacct tccacctaac gttgtatcga
     2521 atttgctttc aatggcccaa caacagcaac aacaacctca tgctcagcag caattggttg
     2581 gtttaataca atcaatgcaa ggccaggctc ctcaacaaca gcaacaacag ttgggtggat
     2641 attcctctat gaactcatcc tctccccctc ctatgagtac caattacaat ggtcaaaata
     2701 tatctgcaaa accctctgcc ccaccaatgt cacaccaacc tccgccacct caacaacaac
     2761 aacaacaaca acaacagcaa cagcaacagc aacagcaacc tgctggcaat aatgttcaaa
     2821 gtctattaga tagtttagca aaactacaaa aatagactcc cttttttcaa tcttttccat
     2881 ttcttgtaca ttatacttga attcttttaa ttggacgaaa aatttttaag gaaagtagat
     2941 tgatatggtt tgcataaatt cgacatagcg gtttatgctt taatcgtttt cccttcccaa
     3001 attggactag gacttaattc ttactattgc tgcatctctt tcagaacgaa aaactcttta
     3061 ccctgttata aatttctttt atttcttcct tatcaaatgc tatttggtct atttatttca
     3121 cttacacgca cctcttcatt tgtttcctct gctcgagggg gggcccggta ccagcttttg
     3181 ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt
     3241 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3301 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3361 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3421 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3481 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3541 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3601 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3661 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3721 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3781 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3841 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3901 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3961 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4021 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4081 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4141 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4201 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4261 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4321 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4381 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4441 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4501 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4561 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4621 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4681 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4741 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4801 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4861 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4921 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4981 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5041 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5101 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5161 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5221 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5281 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5341 ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa
     5401 aataattata atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa
     5461 agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg
     5521 ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt
     5581 tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc
     5641 gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag
     5701 tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact
     5761 cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca
     5821 cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg
     5881 caagcgatcc gtcctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5941 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     6001 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     6061 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     6121 agattgtact gagagtgcac catatcgact acgtcgttaa ggccgtttct gacagagtaa
     6181 aattcttgag ggaactttca ccattatggg aaatggttca agaaggtatt gacttaaact
     6241 ccatcaaatg gtcaggtcat tgagtgtttt ttatttgttg tatttttttt tttttagaga
     6301 aaatcctcca atatataaat taggaatcat agtttcatga ttttctgtta cacctaactt
     6361 tttgtgtggt gccctcctcc ttgtcaatat taatgttaaa gtgcaattct ttttccttat
     6421 cacgttgagc cattagtatc aatttgctta cctgtattcc tttacatcct cctttttctc
     6481 cttcttgata aatgtatgta gattgcgtat atagtttcgt ctaccctatg aacatattcc
     6541 attttgtaat ttcgtgtcgt ttctattatg aatttcattt ataaagttta tgtacaaata
     6601 tcataaaaaa agagaatctt tttaagcaag gattttctta acttcttcgg cgacagcatc
     6661 accgacttcg gtggtactgt tggaaccacc taaatcacca gttctgatac ctgcatccaa
     6721 aaccttttta actgcatctt caatggcctt accttcttca ggcaagttca atgacaattt
     6781 caacatcatt gcagcagaca agatagtggc gatagggttg accttattct ttggcaaatc
     6841 tggagcagaa ccgtggcatg gttcgtacaa accaaatgcg gtgttcttgt ctggcaaaga
     6901 ggccaaggac gcagatggca acaaacccaa ggaacctggg ataacggagg cttcatcgga
     6961 gatgatatca ccaaacatgt tgctggtgat tataatacca tttaggtggg ttgggttctt
     7021 aactaggatc atggcggcag aatcaatcaa ttgatgttga accttcaatg tagggaattc
     7081 gttcttgatg gtttcctcca cagtttttct ccataatctt gaagaggcca aaacattagc
     7141 tttatccaag gaccaaatag gcaatggtgg ctcatgttgt agggccatga aagcggccat
     7201 tcttgtgatt ctttgcactt ctggaacggt gtattgttca ctatcccaag cgacaccatc
     7261 accatcgtct tcctttctct taccaaagta aatacctccc actaattctc tgacaacaac
     7321 gaagtcagta cctttagcaa attgtggctt gattggagat aagtctaaaa gagagtcgga
     7381 tgcaaagtta catggtctta agttggcgta caattgaagt tctttacgga tttttagtaa
     7441 accttgttca ggtctaacac taccggtacc ccatttagga ccacccacag cacctaacaa
     7501 aacggcatca gccttcttgg aggcttccag cgcctcatct ggaagtggaa cacctgtagc
     7561 atcgatagca gcaccaccaa ttaaatgatt ttcgaaatcg aacttgacat tggaacgaac
     7621 atcagaaata gctttaagaa ccttaatggc ttcggctgtg atttcttgac caacgtggtc
     7681 acctggcaaa acgacgatct tcttaggggc agacattaga atggtatatc cttgaaatat
     7741 atatatatat attgctgaaa tgtaaaaggt aagaaaagtt agaaagtaag acgattgcta
     7801 accacctatt ggaaaaaaca ataggtcctt aaataatatt gtcaacttca agtattgtga
     7861 tgcaagcatt tagtcatgaa cgcttctcta ttctatatga aaagccggtt ccggcgctct
     7921 cacctttcct ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct
     7981 gaaattaaca aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg
     8041 ttctcgttat gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta
     8101 cgatacctga gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt
     8161 agaccgctcg gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa
     8221 tataacgttt ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat
     8281 gtggattttg atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt
     8341 agatatacta gaagttctcc tcgaccggtc gatatgcggt gtgaaatacc gcacagatgc
     8401 gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt
     8461 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     8521 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     8581 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     8641 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     8701 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     8761 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     8821 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     8881 ccattcgcca t
//
