LOCUS       sequence-213348-        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   O'Rourke TW, Reines D
  TITLE     Determinants of Amyloid Formation for the Yeast Termination Factor 
            Nab3.
  JOURNAL   PLoS One. 2016 Mar 8;11(3):e0150865. doi: 
            10.1371/journal.pone.0150865. eCollection 2016.
  PUBMED    26954508
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"; journalName: "PLoS One.";
            date: "2016-03-8"; volume: "11(3):e0150865. doi"; pages: " 
            10.1371/journal.pone.0150865. eCollection 2016"
COMMENT     Sequence Label: pRS315Nab3Q->E
FEATURES             Location/Qualifiers
     source          1..8891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..52
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     36..52
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(3016..3034)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3055..3071)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3055..3071)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3068..3090)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3079..3095
                     /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(3103..3109,3110..3127,3128..3133))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3148..3169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3286..3303)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3457..4045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3537..3556)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4216..5007,5008..5076))
                     /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     4839..4858
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5077..5181)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5218..5721
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5763..5781
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5819..5841)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5941..5960
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(6454..7548)
                     /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(7549..7956)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(8257..8712)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8394..8415)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8606..8625
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8838..8860
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8852..8869
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8853..8869
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(8876..8891,1..4)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(8876..8891,1..3)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agggcgaatt ggagctccac cgcggtggcg gccgctctag aactagtgga tccagtgtaa
       61 ccctgaattg ttgaagagaa aatgaagcgg aagaagcaaa ggacaaaaaa caattcattt
      121 gattttgcca cttctaaatg aaggtctaat aaaagctatc ttgagcatct ttattagatt
      181 ctgcacagca acaagcgatt ttctttggtc aaaatataat aattgactta cgtttttccc
      241 ggactgtcct ttcataatat aataaccatc tgcaagccat gtcagatgaa aaccataaca
      301 gtgatgttca agatattcct tcacctgaac tatccgtcga tagtaactct aacgagaatg
      361 aattgatgaa taactcaagc gcagacgatg gaatcgaatt tgacgcccca gaggaagaaa
      421 gagaagccga aagggaggag gaaaatgaag aacaacacga actggaagat gtgaacgatg
      481 aagaggagga agataaggag gaaaaaggag aggaaaacgg ggaagtaata aacacagaag
      541 aagaagaaga agaagaacat caacaaaaag gcggaaatga tgatgacgat gatgataatg
      601 aagaggaaga agaggaagaa gaggatgatg acgatgatga tgatgacgac gatgatgatg
      661 aagaagaaga agaagaagaa gaagaagaag gcaacgacaa cagttcggta ggctcagata
      721 gtgccgctga agacggtgag gatgaggaag acaaaaagga taaaaccaaa gataaagagg
      781 tcgaacttcg ccgtgaaaca ttggaaaaag aacaaaagga cgtagatgaa gctataaaaa
      841 aaataactcg tgaagaaaat gataatactc actttccaac taatatggaa aatgttaatt
      901 acgatctttt acaaaagcaa gtcaagtaca ttatggacag taacatgcta aatttgcctc
      961 agtttcaaca tttacctcaa gaagaaaaga tgtctgcgat tttagcaatg ttaaattcaa
     1021 attctgacac agctctttcc gtacctcctc atgatagtac tatctcaaca acagctagcg
     1081 cctcagccac aagcggcgca agaagcaatg atcaaagaaa acctccattg tcagatgccc
     1141 aaagacgtat gagatttcct agggcggatt tatctaagcc gattaccgaa gaagaacacg
     1201 accgttatgc agcctatttg cacggtgaaa ataaaatcac cgagatgcac aatattcctc
     1261 cgaagtcaag attattcatt ggtaatttgc cgctaaagaa cgtttctaag gaggatttat
     1321 ttaggatttt ctctccatac ggtcatatca tgcaaatcaa tatcaaaaat gcctttggat
     1381 tcattcagtt tgacaaccct caaagcgtta gagatgcaat tgaatgcgag tctcaagaaa
     1441 tgaactttgg caaaaagttg atcctggaag tttctagctc gaatgctcgt cctcaatttg
     1501 atcatggtga tcacggtaca aacagtagtt ctacttttat ttcttccgca aaacgaccat
     1561 ttcaaactga atctggtgac atgtacaatg acgacaatgg tgctggctac aagaaatcca
     1621 gaagacacac cgtttcttgc aacattttcg ttaaaagaac cgcagatcgt acgtatgcca
     1681 ttgaggtttt caacaggttt agggacggga ctggtttgga aactgatatg attttcttga
     1741 aaccaagaat ggaactggga aagcttatca atgatgccgc atataatggg gtgtggggcg
     1801 ttgttttagt taataaaaca cacaatgtag atgttcaaac tttctacaaa ggctcacaag
     1861 gtgaaacgaa atttgatgaa tatattagca tatccgctga tgacgcagtt gccattttta
     1921 ataacatcaa aaacaacaga aataattctc gtcctactga ttaccgtgct atgagccatc
     1981 agcaaaacat atatggcgct cctcctcttc ctgttccaaa cggcccagct gtcggacctc
     2041 ctcctcaaac aaactattac cagggttaca gtatgcctcc tccacaacaa caacagcaac
     2101 agccatatgg taattatggg atgccaccac catcccatga ccaaggatat ggttctcaac
     2161 ctccaattcc aatgaatcag agctacggtc gctaccagac ttctattcca ccaccacctc
     2221 cacaacaaca aattcctcaa gggtatggtc gttatcaggc tggtccgcct cctcaaccac
     2281 cttctgaaac tccaatggac gagcaagaac tattatctgc cattgaaaac cttccaccta
     2341 acgttgtatc gaatttgctt tcaatggccg aacaagagca agaacaacct catgctgagc
     2401 aggaattggt tggtttaata gaatcaatgg aaggcgaggc tcctgaacaa gagcaagaac
     2461 agttgggtgg atattcctct atgaactcat cctctccccc tcctatgagt accaattaca
     2521 atggtgaaaa tatatctgca aaaccctctg ccccaccaat gtcacacgaa cctccgccac
     2581 ctgaacaaca acaagaacaa gaacaagagc aagagcaaga gcaagagcaa cctgctggca
     2641 ataatgttca aagtctatta gatagtttag caaaactaca aaaatagact cccttttttc
     2701 aatcttttcc atttcttgta cattatactt gaattctttt aattggacga aaaattttta
     2761 aggaaagtag attgatatgg tttgcataaa ttcgacatag cggtttatgc tttaatcgtt
     2821 ttcccttccc aaattggact aggacttaat tcttactatt gctgcatctc tttcagaacg
     2881 aaaaactctt taccctgtta taaatttctt ttatttcttc cttatcaaat gctatttggt
     2941 ctatttattt cacttacacg cacctcttca tttgtttcct ctgctcgagg gggggcccgg
     3001 taccagcttt tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata
     3061 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3121 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3181 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3241 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3301 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3361 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3421 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3481 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3541 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3601 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3661 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3721 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3781 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3841 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3901 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3961 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4021 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4081 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4141 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4201 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4261 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4321 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4381 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4441 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4501 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4561 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4621 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4681 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4741 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4801 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4861 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4921 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4981 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5041 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5101 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5161 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt ccttttcatc
     5221 acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa attaaaaata
     5281 gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat gtaaaagact
     5341 ctagggggat cgccaacaaa tactaccttt tatcttgctc ttcctgctct caggtattaa
     5401 tgccgaattg tttcatcttg tctgtgtaga agaccacaca cgaaaatcct gtgattttac
     5461 attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg tctaataaat
     5521 atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt tttttttctt
     5581 cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac aaaacataaa
     5641 aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata cgaggcgcgt
     5701 gtaagttaca ggcaagcgat ccgtcctaag aaaccattat tatcatgaca ttaacctata
     5761 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     5821 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     5881 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     5941 cggcatcaga gcagattgta ctgagagtgc accatatcga ctacgtcgtt aaggccgttt
     6001 ctgacagagt aaaattcttg agggaacttt caccattatg ggaaatggtt caagaaggta
     6061 ttgacttaaa ctccatcaaa tggtcaggtc attgagtgtt ttttatttgt tgtatttttt
     6121 tttttttaga gaaaatcctc caatatataa attaggaatc atagtttcat gattttctgt
     6181 tacacctaac tttttgtgtg gtgccctcct ccttgtcaat attaatgtta aagtgcaatt
     6241 ctttttcctt atcacgttga gccattagta tcaatttgct tacctgtatt cctttacatc
     6301 ctcctttttc tccttcttga taaatgtatg tagattgcgt atatagtttc gtctacccta
     6361 tgaacatatt ccattttgta atttcgtgtc gtttctatta tgaatttcat ttataaagtt
     6421 tatgtacaaa tatcataaaa aaagagaatc tttttaagca aggattttct taacttcttc
     6481 ggcgacagca tcaccgactt cggtggtact gttggaacca cctaaatcac cagttctgat
     6541 acctgcatcc aaaacctttt taactgcatc ttcaatggcc ttaccttctt caggcaagtt
     6601 caatgacaat ttcaacatca ttgcagcaga caagatagtg gcgatagggt tgaccttatt
     6661 ctttggcaaa tctggagcag aaccgtggca tggttcgtac aaaccaaatg cggtgttctt
     6721 gtctggcaaa gaggccaagg acgcagatgg caacaaaccc aaggaacctg ggataacgga
     6781 ggcttcatcg gagatgatat caccaaacat gttgctggtg attataatac catttaggtg
     6841 ggttgggttc ttaactagga tcatggcggc agaatcaatc aattgatgtt gaaccttcaa
     6901 tgtagggaat tcgttcttga tggtttcctc cacagttttt ctccataatc ttgaagaggc
     6961 caaaacatta gctttatcca aggaccaaat aggcaatggt ggctcatgtt gtagggccat
     7021 gaaagcggcc attcttgtga ttctttgcac ttctggaacg gtgtattgtt cactatccca
     7081 agcgacacca tcaccatcgt cttcctttct cttaccaaag taaatacctc ccactaattc
     7141 tctgacaaca acgaagtcag tacctttagc aaattgtggc ttgattggag ataagtctaa
     7201 aagagagtcg gatgcaaagt tacatggtct taagttggcg tacaattgaa gttctttacg
     7261 gatttttagt aaaccttgtt caggtctaac actaccggta ccccatttag gaccacccac
     7321 agcacctaac aaaacggcat cagccttctt ggaggcttcc agcgcctcat ctggaagtgg
     7381 aacacctgta gcatcgatag cagcaccacc aattaaatga ttttcgaaat cgaacttgac
     7441 attggaacga acatcagaaa tagctttaag aaccttaatg gcttcggctg tgatttcttg
     7501 accaacgtgg tcacctggca aaacgacgat cttcttaggg gcagacatta gaatggtata
     7561 tccttgaaat atatatatat atattgctga aatgtaaaag gtaagaaaag ttagaaagta
     7621 agacgattgc taaccaccta ttggaaaaaa caataggtcc ttaaataata ttgtcaactt
     7681 caagtattgt gatgcaagca tttagtcatg aacgcttctc tattctatat gaaaagccgg
     7741 ttccggcgct ctcacctttc ctttttctcc caatttttca gttgaaaaag gtatatgcgt
     7801 caggcgacct ctgaaattaa caaaaaattt ccagtcatcg aatttgattc tgtgcgatag
     7861 cgcccctgtg tgttctcgtt atgttgagga aaaaaataat ggttgctaag agattcgaac
     7921 tcttgcatct tacgatacct gagtattccc acagttaact gcggtcaaga tatttcttga
     7981 atcaggcgcc ttagaccgct cggccaaaca accaattact tgttgagaaa tagagtataa
     8041 ttatcctata aatataacgt ttttgaacac acatgaacaa ggaagtacag gacaattgat
     8101 tttgaagaga atgtggattt tgatgtaatt gttgggattc catttttaat aaggcaataa
     8161 tattaggtat gtagatatac tagaagttct cctcgaccgg tcgatatgcg gtgtgaaata
     8221 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt
     8281 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     8341 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     8401 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     8461 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     8521 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     8581 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     8641 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     8701 tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     8761 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     8821 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata
     8881 cgactcacta t
//
