LOCUS       sequence-213344-        8031 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 8031)
  AUTHORS   Loya TJ, O'Rourke TW, Reines D
  TITLE     The hnRNP-like Nab3 termination factor can employ heterologous 
            prion-like domains in place of its own essential low complexity 
            domain.
  JOURNAL   PLoS One. 2017 Oct 12;12(10):e0186187. doi: 
            10.1371/journal.pone.0186187. eCollection 2017.
  PUBMED    29023495
REFERENCE   2  (bases 1 to 8031)
  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"; journalName: "PLoS One.";
            date: "2017-10-12"; volume: "12(10):e0186187. doi"; pages: " 
            10.1371/journal.pone.0186187. eCollection 2017"
COMMENT     Sequence Label: pRS315Nab3D191stop
FEATURES             Location/Qualifiers
     source          1..8031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3..21
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     54..70
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     54..70
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2174..2192)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2213..2229)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2213..2229)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2226..2248)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2237..2253
                     /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(2261..2267,2268..2285,2286..2291))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2306..2327
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2444..2461)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2615..3203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2695..2714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3374..4165,4166..4234))
                     /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     3997..4016
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4235..4339)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4376..4879
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4921..4939
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4977..4999)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5099..5118
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(5612..6706)
                     /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(6707..7114)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(7415..7870)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7552..7573)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7764..7783
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7996..8018
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8010..8027
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8011..8027
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgtaatacga ctcactatag ggcgaattgg agctccaccg cggtggcggc cgctctagaa
       61 ctagtggatc cagtgtaacc ctgaattgtt gaagagaaaa tgaagcggaa gaagcaaagg
      121 acaaaaaaca attcatttga ttttgccact tctaaatgaa ggtctaataa aagctatctt
      181 gagcatcttt attagattct gcacagcaac aagcgatttt ctttggtcaa aatataataa
      241 ttgacttacg tttttcccgg actgtccttt cataatataa taaccatctg caagccatgt
      301 cagatgaaaa ccataacagt gatgttcaag atattccttc acctgaacta tccgtcgata
      361 gtaactctaa cgagaatgaa ttgatgaata actcaagcgc agacgatgga atcgaatttg
      421 acgccccaga ggaagaaaga gaagccgaaa gggaggagga aaatgaagaa caacacgaac
      481 tggaagatgt gaacgatgaa gaggaggaag ataaggagga aaaaggagag gaaaacgggg
      541 aagtaataaa cacagaagaa gaagaagaag aagaacatca acaaaaaggc ggaaatgatg
      601 atgacgatga tgataatgaa gaggaagaag aggaagaaga ggatgatgac gatgatgatg
      661 atgacgacga tgatgatgaa gaagaagaag aagaagaaga agaagaaggc aacgacaaca
      721 gttcggtagg ctcagatagt gccgctgaag acggtgagga tgaggaagac aaaaaggata
      781 aaaccaaaga taaagaggtc gaacttcgcc gtgaaacatt ggaaaaagaa caaaaggacg
      841 tagatgaagc tataaaaaaa ataactcgtg aagaaaatga taatactcac tttccaacta
      901 atatggaaaa tgttaattac gatcttttac aaaagcaagt caagtacatt atggacagta
      961 acatgctaaa tttgcctcag tttcaacatt tacctcaaga agaaaagatg tctgcgattt
     1021 tagcaatgtt aaattcaaat tctgacacag ctctttccgt acctcctcat gatagtacta
     1081 tctcaacaac agctagcgcc tcagccacaa gcggcgcaag aagcaatgat caaagaaaac
     1141 ctccattgtc agatgcccaa agacgtatga gatttcctag ggcggattta tctaagccga
     1201 ttaccgaaga agaacacgac cgttatgcag cctatttgca cggtgaaaat aaaatcaccg
     1261 agatgcacaa tattcctccg aagtcaagat tattcattgg taatttgccg ctaaagaacg
     1321 tttctaagga ggatttattt aggattttct ctccatacgg tcatatcatg caaatcaata
     1381 tcaaaaatgc ctttggattc attcagtttg acaaccctca aagcgttaga gatgcaattg
     1441 aatgcgagtc tcaagaaatg aactttggca aaaagttgat cctggaagtt tctagctcga
     1501 atgctcgtcc tcaatttgat catggtgatc acggtacaaa cagtagttct acttttattt
     1561 cttccgcaaa acgaccattt caaactgaat ctggtgacat gtacaatgac gacaatggtg
     1621 ctggctacaa gaaatccaga agacacaccg tttcttgcaa cattttcgtt aaaagaaccg
     1681 cagatcgtac gtatgccatt gaggttttca acaggtttag ggacgggact ggtttggaaa
     1741 ctgatatgat tttcttgaaa ccaagaatgg aactgggaaa gcttatcaat gatgccgcat
     1801 ataatggggt gtggggcgtt gttttagtta ataaaacaca caatgtagat gttcaaactt
     1861 tctacaaagg ctcacaaggt gaaacgaaat ttgatgaata tattagcata tccgctgatg
     1921 acgcagttgc catttttaat aacatcaaaa acaacagaaa taattctcgt cctactgatt
     1981 accgtgctat gagccatcag caaaacatat atggcgctcc tcctcttcct gttccaaacg
     2041 gcccagctgt cggacctcct cctcaaacaa actattacca gggttacagt atgcctcctc
     2101 cacaacaaca acagcaacag ccatattagc catggcccgg gctcgagggg gggcccggta
     2161 ccagcttttg ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc
     2221 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     2281 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2341 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2401 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2461 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2521 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2581 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2641 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2701 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2761 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2821 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2881 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2941 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3001 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3061 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3121 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3181 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3241 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3301 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3361 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3421 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3481 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3541 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3601 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3661 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3721 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3781 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3841 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3901 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3961 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4021 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4081 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4141 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4201 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4261 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4321 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac
     4381 gtgctataaa aataattata atttaaattt tttaatataa atatataaat taaaaataga
     4441 aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct
     4501 agggggatcg ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg
     4561 ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat
     4621 tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat
     4681 atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca
     4741 ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa
     4801 taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt
     4861 aagttacagg caagcgatcc gtcctaagaa accattatta tcatgacatt aacctataaa
     4921 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     4981 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     5041 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     5101 gcatcagagc agattgtact gagagtgcac catatcgact acgtcgttaa ggccgtttct
     5161 gacagagtaa aattcttgag ggaactttca ccattatggg aaatggttca agaaggtatt
     5221 gacttaaact ccatcaaatg gtcaggtcat tgagtgtttt ttatttgttg tatttttttt
     5281 tttttagaga aaatcctcca atatataaat taggaatcat agtttcatga ttttctgtta
     5341 cacctaactt tttgtgtggt gccctcctcc ttgtcaatat taatgttaaa gtgcaattct
     5401 ttttccttat cacgttgagc cattagtatc aatttgctta cctgtattcc tttacatcct
     5461 cctttttctc cttcttgata aatgtatgta gattgcgtat atagtttcgt ctaccctatg
     5521 aacatattcc attttgtaat ttcgtgtcgt ttctattatg aatttcattt ataaagttta
     5581 tgtacaaata tcataaaaaa agagaatctt tttaagcaag gattttctta acttcttcgg
     5641 cgacagcatc accgacttcg gtggtactgt tggaaccacc taaatcacca gttctgatac
     5701 ctgcatccaa aaccttttta actgcatctt caatggcctt accttcttca ggcaagttca
     5761 atgacaattt caacatcatt gcagcagaca agatagtggc gatagggttg accttattct
     5821 ttggcaaatc tggagcagaa ccgtggcatg gttcgtacaa accaaatgcg gtgttcttgt
     5881 ctggcaaaga ggccaaggac gcagatggca acaaacccaa ggaacctggg ataacggagg
     5941 cttcatcgga gatgatatca ccaaacatgt tgctggtgat tataatacca tttaggtggg
     6001 ttgggttctt aactaggatc atggcggcag aatcaatcaa ttgatgttga accttcaatg
     6061 tagggaattc gttcttgatg gtttcctcca cagtttttct ccataatctt gaagaggcca
     6121 aaacattagc tttatccaag gaccaaatag gcaatggtgg ctcatgttgt agggccatga
     6181 aagcggccat tcttgtgatt ctttgcactt ctggaacggt gtattgttca ctatcccaag
     6241 cgacaccatc accatcgtct tcctttctct taccaaagta aatacctccc actaattctc
     6301 tgacaacaac gaagtcagta cctttagcaa attgtggctt gattggagat aagtctaaaa
     6361 gagagtcgga tgcaaagtta catggtctta agttggcgta caattgaagt tctttacgga
     6421 tttttagtaa accttgttca ggtctaacac taccggtacc ccatttagga ccacccacag
     6481 cacctaacaa aacggcatca gccttcttgg aggcttccag cgcctcatct ggaagtggaa
     6541 cacctgtagc atcgatagca gcaccaccaa ttaaatgatt ttcgaaatcg aacttgacat
     6601 tggaacgaac atcagaaata gctttaagaa ccttaatggc ttcggctgtg atttcttgac
     6661 caacgtggtc acctggcaaa acgacgatct tcttaggggc agacattaga atggtatatc
     6721 cttgaaatat atatatatat attgctgaaa tgtaaaaggt aagaaaagtt agaaagtaag
     6781 acgattgcta accacctatt ggaaaaaaca ataggtcctt aaataatatt gtcaacttca
     6841 agtattgtga tgcaagcatt tagtcatgaa cgcttctcta ttctatatga aaagccggtt
     6901 ccggcgctct cacctttcct ttttctccca atttttcagt tgaaaaaggt atatgcgtca
     6961 ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg
     7021 cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg ttgctaagag attcgaactc
     7081 ttgcatctta cgatacctga gtattcccac agttaactgc ggtcaagata tttcttgaat
     7141 caggcgcctt agaccgctcg gccaaacaac caattacttg ttgagaaata gagtataatt
     7201 atcctataaa tataacgttt ttgaacacac atgaacaagg aagtacagga caattgattt
     7261 tgaagagaat gtggattttg atgtaattgt tgggattcca tttttaataa ggcaataata
     7321 ttaggtatgt agatatacta gaagttctcc tcgaccggtc gatatgcggt gtgaaatacc
     7381 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta
     7441 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     7501 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     7561 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     7621 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     7681 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     7741 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     7801 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     7861 cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     7921 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     7981 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa t
//
