LOCUS       sequence-500499-        9108 bp    DNA     circular SYN 19-FEB-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9108)
  AUTHORS   Malagon F, Tong AH, Shafer BK, Strathern JN
  TITLE     Genetic interactions of DST1 in Saccharomyces cerevisiae suggest a 
            role of TFIIS in the initiation-elongation transition.
  JOURNAL   Genetics 2004 Mar;166(3):1215-27.
  PUBMED    15082542
REFERENCE   2  (bases 1 to 9108)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 19, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics"; 
            date: "2004-03"; volume: "166(3)"; pages: "1215-27"
COMMENT     Sequence Label: YEplac181-RTS1
FEATURES             Location/Qualifiers
     source          1..9108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3659..3676)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3659..3675)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3668..3690)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4112..5458
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(5596..6690)
                     /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(6691..7096)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(7116..7134)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7202..7306
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7307..7375,7376..8167)
                     /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     complement(7525..7544)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8338..8926
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8827..8846
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9080..9097
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgactcta gaggatcccc cgaagatata tttggagaaa agtatagaaa
      301 taataaaatg aagacacatc gattgaagta tgctcaaatg atgagaaaaa gagaccttta
      361 tctcaggcca caaacaacca ttgtccaagt ggcttatctt tgctatagtt ttgttcgaat
      421 attcggggat tctatctttg gtttcttcaa caggttgacc ccatttacca gttctttttt
      481 gctcttcata tactgctgct cttttgaaat gtggcatgcc ctaaacttcc tcacttcttc
      541 gagcttgtaa tgaattgctg tttcactgta tctcgctgtc acaatcattt tcgttctctt
      601 catttaatgt gttattctca cttgctgtgt tgaaactgtt tcttaattct gggtttttga
      661 ttgtgtattg gtccttatca ttgttaatcc ttagattttt tacatattct tcaagtttgc
      721 tccaattttc ttcacgttgc accttacgct gttgagtttc ttttacactt gataagtaca
      781 atgcattaca gttttcgtac aatactgggt ttgtttccag aaaaattttc aatgccttga
      841 atgccatagc atgaattgcc ctattccacg aaccagaatt gattgcttgc tcaacaagca
      901 tgtaagggtc tgaaatgcta tcttcgccat ttgctgtatc tagctctaac tgagaagtta
      961 attcatataa tgcaggaaat ataatgggta ggatgacttc ggcattttcg atacataagt
     1021 ttaagaaata ttcattattc caataactta aaaccttttc cgccacttgg aaatgtggag
     1081 aagaaataca cttagctaat tgaacaaata acggaacttc tactttaata aattccagcg
     1141 gttcgatcac ttcaaaaata tcctcgattt catttagaaa cattatctct tttgtggaat
     1201 ttatttttgg ccaataacgc agtaagccca taactacctc ttcggttaat aaaggatctt
     1261 tttcgagaaa ttgaacgata cagtaagcca actgagggtg gtataatgat aaacaacgga
     1321 ccttgtgtaa tggtatcaat atcctcacca agaaaacctt gtgctcttcc tttaatggaa
     1381 gtgcaaatcc attaattatg gaacccaaaa tttctaacaa ttctgccaca ccgttaaact
     1441 tctcagtctc ataaataaat tgcaaaaaaa tattattcat cgaccgacga ataaagcttc
     1501 ttaatgataa gaacttccca tatattctat gcagtgtcgt tttcaaacag tctctttctc
     1561 tgatatcttc gctatcaaat aattccagta actttaaaat aaagtcctga tcaatatatt
     1621 gtttagcaat ctgatgattg aaatcaggac tttccacaaa ccttaaaaag aattcgtaaa
     1681 cagcttgcat atgaggccag gctagttcgt taacaggttc atcttcatct gggtcataaa
     1741 tgtcaccaac tggatttact ggtggtggaa taggtctaaa cagattgatt ttgaacatgt
     1801 tcaccacatg agcgtacatc tcattcgtgt aagtgaacct atttgttaca aggaattcta
     1861 ttagctcatc taaggtgctt cttttaatct ctttaccttg aatgtcaaaa cttggatcat
     1921 taaagtcaaa catagtgtta cactggtcaa ctttggcaat gaacaaaggg attctttctt
     1981 caggagaaac ttcattaaaa tttggcaatt tcgttaatgg cgtatatcta gatggctcga
     2041 atctagatga agaatgacgt tggggagtct taattaactc taaatctgtg tcagggttaa
     2101 gttttgtggg tgttggtagt ctctcaaatg agtgtgatga tctcggaatg tcgatacttt
     2161 gtgaggaggc gtggtttgca tcctgagtat tactagtatt aatattcata ttgttattat
     2221 tactattacc attttctaca tcagaagaaa ctgctgtacc tgaagcattt gaaatgggag
     2281 atgatggaac agcggatact gcccccagca aagtaggatg tgcatgatct tcaccagaag
     2341 tgcttttgtc gtccttagta acagtgagta ctggagtggg gttcattata gcggcagatg
     2401 atgaagaaga tccttgcttt tgagattgtg atggctgttg cgattgtttg gactgttctt
     2461 gtccttttct tgaacttgtt ttctttgtgg agcttgaacc ggagcgactg acacttgaac
     2521 ttctactgct actgctgctg gaagaagggg tcgtcgctgt ttttgttttt gatgtagagc
     2581 tactatgatg cttcccttgc gagggaacac taccagattg cttgcccttc tcagttgtag
     2641 actttgatcc ggtactgctg gcagaactgt gagtagtccc gtgggaagag ctactagccg
     2701 aagcgggctt ctttgatgtg gatgaggtag tggaactttt ttctttttcc ttctctttgt
     2761 ccttcttttt actgcttgaa gaagaagaag acccggtggt cttctttatt aatctttgct
     2821 tgaaaccacg catcatattg aaagtggatg ttcgaaaata gcacgtgcct attgatctta
     2881 ataagtagaa cttttataca aaataaaaaa gccagttaaa gacggtattg aggtgaacag
     2941 gtatatggag aagggtttat atttttatct aacaaaaaaa ggaaaaaaaa gcaaaatcga
     3001 ggggggaagg aaatgaaacc caaaatactt acggcgacga tggtttatcc ttgaaatgaa
     3061 gagttttcca atgaagaaac gagacgtatt gacatgccta ccaattgaag tggaaaacaa
     3121 agcgtagttt cacatactct ttgataatca attgtctgat actttttttt ttcctttctc
     3181 gcagccggga tagggggttg tgacccggca aaaaatgttg gtctaagttt tattgttgct
     3241 atatgctcca tacatacaat aaatgtctat aaaattttta gaagaaaatc aggcgtgaaa
     3301 gatggtgtct gcatctttgt ggtagagcat caagtaggcc gtgaatccct tcccctgcgg
     3361 atcgggttct cctttggagt tcacttctcc aaccccgaac tgaaacacca aataggtttt
     3421 gaaaccgaaa tattcgtaca cagatcttgc gtgctccgaa atagcctcga gagcaagggc
     3481 gcaattggcc ttgtctgctt tggttttata tttttcgagt atttttctga ctgaacctct
     3541 aatttcggga tggctaggat cttttccgat ggcataaagg tagtaatagt ccattcccag
     3601 aggcatgact tcgtgcttcc tcacagtcaa gtcgtgaaag ggtaccgagc tcgaattcac
     3661 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     3721 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3781 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     3841 cgcatctgtg cggtatttca caccgcattt atcggatcgt acttgttacc catcattgaa
     3901 ttttgaacat ccgaacctgg gagttttccc tgaaacagat agtatatttg aacctgtata
     3961 ataatatata gtctagcgct ttacggaaga caatgtatgt atttcggttc ctggagaaac
     4021 tattgcatct attgcatagg taatcttgca cgtcgcatcc ccggttcatt ttctgcgttt
     4081 ccatcttgca cttcaatagc atatctttgt taacgaagca tctgtgcttc attttgtaga
     4141 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     4201 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     4261 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     4321 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     4381 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     4441 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     4501 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     4561 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     4621 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     4681 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     4741 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     4801 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     4861 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     4921 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     4981 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     5041 tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     5101 aataggaact tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg
     5161 caacgcgagc tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc
     5221 tgtatatata tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt
     5281 atatgcgtct atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat
     5341 tccatgcggg gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc
     5401 actcctcaat tggattagtc tcatccttca atgctatcat ttcctttgat attggatcga
     5461 tccgatgata agctgtcaaa catgagaatt aattctaccc tatgaacata ttccattttg
     5521 taatttcgtg tcgtttctat tatgaatttc atttataaag tttatgtaca aatatcataa
     5581 aaaaagagaa tctttttaag caaggatttt cttaacttct tcggcgacag catcaccgac
     5641 ttcggtggta ctgttggaac cacctaaatc accagttctg atacctgcat ccaaaacctt
     5701 tttaactgca tcttcaatgg ccttaccttc ttcaggcaag ttcaatgaca atttcaacat
     5761 cattgcagca gacaagatag tggcgatagg gttgacctta ttctttggca aatctggagc
     5821 agaaccgtgg catggttcgt acaaaccaaa tgcggtgttc ttgtctggca aagaggccaa
     5881 ggacgcagat ggcaacaaac ccaaggaacc tgggataacg gaggcttcat cggagatgat
     5941 atcaccaaac atgttgctgg tgattataat accatttagg tgggttgggt tcttaactag
     6001 gatcatggcg gcagaatcaa tcaattgatg ttgaaccttc aatgtaggaa attcgttctt
     6061 gatggtttcc tccacagttt ttctccataa tcttgaagag gccaaaacat tagctttatc
     6121 caaggaccaa ataggcaatg gtggctcatg ttgtagggcc atgaaagcgg ccattcttgt
     6181 gattctttgc acttctggaa cggtgtattg ttcactatcc caagcgacac catcaccatc
     6241 gtcttccttt ctcttaccaa agtaaatacc tcccactaat tctctgacaa caacgaagtc
     6301 agtaccttta gcaaattgtg gcttgattgg agataagtct aaaagagagt cggatgcaaa
     6361 gttacatggt cttaagttgg cgtacaattg aagttcttta cggattttta gtaaaccttg
     6421 ttcaggtcta acactacctg taccccattt aggaccaccc acagcaccta acaaaacggc
     6481 atcagccttc ttggaggctt ccagcgcctc atctggaagt ggaacacctg tagcatcgat
     6541 agcagcacca ccaattaaat gattttcgaa atcgaacttg acattggaac gaacatcaga
     6601 aatagcttta agaaccttaa tggcttcggc tgtgatttct tgaccaacgt ggtcacctgg
     6661 caaaacgacg atcttcttag gggcagacat tagaatggta tatccttgaa atatatatat
     6721 atattgctga aatgtaaaag gtaagaaaag ttagaaagta agacgattgc taaccaccta
     6781 ttggaaaaaa caataggtcc ttaaataata ttgtcaactt caagtattgt gatgcaagca
     6841 tttagtcatg aacgcttctc tattctatat gaaaagccgg ttccggcgct ctcacctttc
     6901 ctttttctcc caatttttca gttgaaaaag gtatatgcgt caggcgacct ctgaaattaa
     6961 caaaaaattt ccagtcatcg aatttgattc tgtgcgatag cgcccctgtg tgttctcgtt
     7021 atgttgagga aaaaaataat ggttgctaag agattcgaac tcttgcatct tacgatacct
     7081 gagtattccc acagttaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     7141 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     7201 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     7261 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     7321 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     7381 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     7441 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     7501 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     7561 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     7621 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     7681 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     7741 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     7801 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     7861 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     7921 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7981 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     8041 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     8101 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     8161 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     8221 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     8281 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     8341 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     8401 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     8461 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     8521 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     8581 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     8641 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     8701 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     8761 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     8821 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     8881 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     8941 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     9001 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     9061 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaaga
//
