LOCUS       sequence-402638-        7850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7850)
  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 7850)
  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: "Genetics"; 
            date: "2004-03"; volume: "166(3)"; pages: "1215-27"
COMMENT     Sequence Label: YEplac181-TYS1
FEATURES             Location/Qualifiers
     source          1..7850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     369..386
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    503..524
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(539..544,545..562,563..569)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    577..593
                     /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     582..604
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     601..617
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     601..617
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2798..2815)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2798..2814)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2807..2829)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3251..4597
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4735..5829)
                     /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(5830..6235)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(6255..6273)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6341..6445
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6446..6514,6515..7306)
                     /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(6664..6683)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7477..7850,1..215)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
       61 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      301 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      361 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      421 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      481 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      541 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      601 caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcagcat cggcctcgac
      661 accgatttgc aaaagcattt tctctctctc ttcagcatac tcaggatcat cctttttgac
      721 aagagatccc ctgctgttgt ctccatttgg tacgcctggg gaactagaat tgctgccgtt
      781 ttgacccatt ctcatcctca ggaaatttct tctcattctg tctcttttaa tgaagggaga
      841 ggcattcgtc ggacctcccc cgtttctact gcctggtgga ttgcgtctgg acatccttgc
      901 gcttactcga ataggcctcc ctagctattc ttcaaccttt cgaaccatcc atacttctta
      961 ctatcataat ttttatttta tcatggaggc gagaaggtcc ttattcgagc atcactaaga
     1021 acggaactcg aacatttaca aagtagaaaa attttatgaa aattaattgt tctttcttca
     1081 gaatacaaat tagtcattgt caaaaagaga ttagcatcca taaccgcata ctctaattga
     1141 cgataacatg tcctctgctg ccacggttga ccctaatgag gcgttcggtc tcattaccaa
     1201 gaacttgcaa gaagtcttga acccacaaat catcaaagat gttctcgaag tacaaaagag
     1261 acatttgaaa ttatactggg gtaccgcgcc cactggaaga cctcattgtg gttattttgt
     1321 tcctatgacc aagcttgctg atttcttgaa agcaggttgt gaagtgactg ttcttttagc
     1381 agacttgcac gcctttttgg acaacatgaa ggcgccattg gaagtggtga actatagggc
     1441 caaatactat gaattgacca tcaaagctat cttgagaagc atcaatgttc caattgaaaa
     1501 gctaaaattt gttgttggct cttcttacca gctaactcca gactatacca tggatatctt
     1561 tagattatcc aacattgttt cacaaaacga tgctaaaagg gcaggcgctg atgttgtcaa
     1621 gcaagtcgcc aatccattat tgagtgggct tatctatcct ttaatgcaag cgttggacga
     1681 acaattctta gatgttgatt gccaatttgg tggtgtcgac cagagaaaaa tttttgtctt
     1741 agcggaagaa aatttaccaa gtttgggcta caagaaaaga gcacatttaa tgaaccctat
     1801 ggttccaggt ttggctcaag gtggtaaaat gtctgcctct gatccgaatt caaaaatcga
     1861 ccttttggaa gaaccaaagc aagttaagaa aaagatcaat agtgcatttt gtagccccgg
     1921 taacgtcgaa gagaatggtt tactgtcatt tgtccaatat gtcattgcac caatccaaga
     1981 attgaagttt ggtacaaatc acttcgaatt cttcattgat agaccagaaa agttcggtgg
     2041 tccaattacc tacaaatcct tcgaagaaat gaaattggct tttaaagaag aaaaattgtc
     2101 cccacctgat ctaaaaattg gtgttgctga cgctatcaac gagctattgg agccaatcag
     2161 acaagaattt gccaacaaca aggaatttca agaggcctcc gaaaagggtt atccagttgc
     2221 cacaccacaa aagtcgaaaa aggctaagaa accaaagaat aagggtacta agtacccagg
     2281 cgctactaag accaacgaga ttgccacgaa actagaggaa accaaattgt aagcatagct
     2341 taatccgttt tcacgattca taatataata aataagaaaa gatatatcat ataaacgtta
     2401 taaaattaat aaccgggtaa gtgtagaaaa gtgatgcgac ggtttatttt ctcttcctct
     2461 tgcgattgaa tttaacttgc agatagtgac cataaggcaa ctacccagtg gcaaacagtt
     2521 ttgataacgc ccagtacatc aacgagcgag tataaagact ttggtacatt ttaaaaagga
     2581 aacatatatt gttttcattg ctagaccctt ttagtctcac ctcaataaaa ctgctttatt
     2641 cctcattggg ctttttattc tttaattttg catacttata gcgtgaaact gggcatttaa
     2701 caaaagcaaa ctattttaat agtagcatcc tgctttcttt gcccctcctt cttattgcga
     2761 tacattatta agttttttta ccaccgagct cgaattcact ggccgtcgtt ttacaacgtc
     2821 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     2881 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     2941 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3001 accgcattta tcggatcgta cttgttaccc atcattgaat tttgaacatc cgaacctggg
     3061 agttttccct gaaacagata gtatatttga acctgtataa taatatatag tctagcgctt
     3121 tacggaagac aatgtatgta tttcggttcc tggagaaact attgcatcta ttgcataggt
     3181 aatcttgcac gtcgcatccc cggttcattt tctgcgtttc catcttgcac ttcaatagca
     3241 tatctttgtt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     3301 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     3361 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     3421 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3481 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     3541 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     3601 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     3661 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     3721 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     3781 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     3841 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     3901 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     3961 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     4021 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     4081 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     4141 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt
     4201 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cggaatagga
     4261 acttcaaagc gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac
     4321 agctcactgt tcacgtcgca cctatatctg cgtgttgcct gtatatatat atacatgaga
     4381 agaacggcat agtgcgtgtt tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg
     4441 atgaaaggta gtctagtacc tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc
     4501 ttccttcagc actacccttt agctgttcta tatgctgcca ctcctcaatt ggattagtct
     4561 catccttcaa tgctatcatt tcctttgata ttggatcgat ccgatgataa gctgtcaaac
     4621 atgagaatta attctaccct atgaacatat tccattttgt aatttcgtgt cgtttctatt
     4681 atgaatttca tttataaagt ttatgtacaa atatcataaa aaaagagaat ctttttaagc
     4741 aaggattttc ttaacttctt cggcgacagc atcaccgact tcggtggtac tgttggaacc
     4801 acctaaatca ccagttctga tacctgcatc caaaaccttt ttaactgcat cttcaatggc
     4861 cttaccttct tcaggcaagt tcaatgacaa tttcaacatc attgcagcag acaagatagt
     4921 ggcgataggg ttgaccttat tctttggcaa atctggagca gaaccgtggc atggttcgta
     4981 caaaccaaat gcggtgttct tgtctggcaa agaggccaag gacgcagatg gcaacaaacc
     5041 caaggaacct gggataacgg aggcttcatc ggagatgata tcaccaaaca tgttgctggt
     5101 gattataata ccatttaggt gggttgggtt cttaactagg atcatggcgg cagaatcaat
     5161 caattgatgt tgaaccttca atgtaggaaa ttcgttcttg atggtttcct ccacagtttt
     5221 tctccataat cttgaagagg ccaaaacatt agctttatcc aaggaccaaa taggcaatgg
     5281 tggctcatgt tgtagggcca tgaaagcggc cattcttgtg attctttgca cttctggaac
     5341 ggtgtattgt tcactatccc aagcgacacc atcaccatcg tcttcctttc tcttaccaaa
     5401 gtaaatacct cccactaatt ctctgacaac aacgaagtca gtacctttag caaattgtgg
     5461 cttgattgga gataagtcta aaagagagtc ggatgcaaag ttacatggtc ttaagttggc
     5521 gtacaattga agttctttac ggatttttag taaaccttgt tcaggtctaa cactacctgt
     5581 accccattta ggaccaccca cagcacctaa caaaacggca tcagccttct tggaggcttc
     5641 cagcgcctca tctggaagtg gaacacctgt agcatcgata gcagcaccac caattaaatg
     5701 attttcgaaa tcgaacttga cattggaacg aacatcagaa atagctttaa gaaccttaat
     5761 ggcttcggct gtgatttctt gaccaacgtg gtcacctggc aaaacgacga tcttcttagg
     5821 ggcagacatt agaatggtat atccttgaaa tatatatata tattgctgaa atgtaaaagg
     5881 taagaaaagt tagaaagtaa gacgattgct aaccacctat tggaaaaaac aataggtcct
     5941 taaataatat tgtcaacttc aagtattgtg atgcaagcat ttagtcatga acgcttctct
     6001 attctatatg aaaagccggt tccggcgctc tcacctttcc tttttctccc aatttttcag
     6061 ttgaaaaagg tatatgcgtc aggcgacctc tgaaattaac aaaaaatttc cagtcatcga
     6121 atttgattct gtgcgatagc gcccctgtgt gttctcgtta tgttgaggaa aaaaataatg
     6181 gttgctaaga gattcgaact cttgcatctt acgatacctg agtattccca cagttaattc
     6241 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     6301 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     6361 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6421 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6481 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6541 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6601 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6661 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6721 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6781 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6841 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6901 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6961 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     7021 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     7081 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     7141 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     7201 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     7261 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     7321 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     7381 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7441 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7501 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7561 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7621 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7681 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7741 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7801 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
//
