LOCUS       sequence-271631-        7029 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 7029)
  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 7029)
  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: pRS415-SOH1
FEATURES             Location/Qualifiers
     source          1..7029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(56..511)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(193..214)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     405..424
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     637..659
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     651..668
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     652..668
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     678..697
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        678..696
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1844..1864)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1844..1862)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1883..1899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1883..1899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1896..1918)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1907..1923
                     /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(1931..1937,1938..1955,1956..1961))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1976..1997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2114..2131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2285..2873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2365..2384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3044..3835,3836..3904))
                     /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     3667..3686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3905..4009)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4046..4549
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4591..4609
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4647..4669)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4769..4788
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(5282..6376)
                     /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(6377..6784)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
ORIGIN
        1 cgatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
       61 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
      121 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
      181 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
      241 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
      301 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
      361 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
      421 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
      481 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
      541 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
      601 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
      661 cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc
      721 tcgagatctc taatgtcttg cgagataata tgcaaggaaa acaaaggaaa gaaaatacaa
      781 agtaatgcgg cttgcatata tatatacata tatatatgta tatatgtgat atatatatgt
      841 gtgtgtgtgt atatatattc atgagtgtat aaaatagata tatgaatgcg tgcgaatgta
      901 gttataatta ggtgtgttca gttggcccat ctttctacca tttcattcat ccattgcggg
      961 ccttgtaatt gaatgatctt gggcagctcg tctagccctt ccagcaggcc atcttcatta
     1021 acgatcgccg actccatgaa cccgttcaac agctttaaaa tgaagagaca gtttggatac
     1081 acaatgcatt gcgaataagg ggggttgcac cagtactcca ggtatttcaa gtagtttttg
     1141 aagttaggag acttccagat ctgctgttga gtgagtaagt acgttacgta ttggatgttt
     1201 gccagtgact ggataaattc gagctccact tcgaacctcg tggggagagg attttgatcc
     1261 gaggaaggtg tcgctggtgc gtttccatta gtgcttgaca tcctgggtgg agcttggggg
     1321 aactttgccg tcttatgagt gctataagtt ttggcttact ttgcctcatt tttcctgcat
     1381 ttctttttct ttttcatcct ttaagaacac gttagcggga aaaccgtgaa ataaaataca
     1441 aaacgaactg ttcgaaaaaa gccaaagggg aaacgcctaa aactccactg aaaccagcta
     1501 taaagcacgg catattacaa caatgtctag caaatggttt aatgctatac acctactggt
     1561 gtgcccgttg acggtactgg taggatatct catgaacgcg tatggctacg gtgcggcgct
     1621 gcaagcaacc ctgaataagg atggtctggt aaatgctatg ttggtaaaga aagggtggtt
     1681 ttggacttcc ttggtcggat ggtggtgtat tatacgctac cgtgcggtgc cgggggcaac
     1741 cggcagagac cgaagacaca ttgtccagtc attcaaaagg tatgccatac tgacagtgtg
     1801 gtctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag tgagggttaa
     1861 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1921 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1981 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2041 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3061 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3661 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4021 gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt
     4081 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
     4141 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
     4201 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
     4261 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
     4321 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
     4381 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
     4441 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
     4501 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcctaagaa
     4561 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     4621 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     4681 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     4741 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     4801 catatcgact acgtcgttaa ggccgtttct gacagagtaa aattcttgag ggaactttca
     4861 ccattatggg aaatggttca agaaggtatt gacttaaact ccatcaaatg gtcaggtcat
     4921 tgagtgtttt ttatttgttg tatttttttt tttttagaga aaatcctcca atatataaat
     4981 taggaatcat agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
     5041 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
     5101 aatttgctta cctgtattcc tttacatcct cctttttctc cttcttgata aatgtatgta
     5161 gattgcgtat atagtttcgt ctaccctatg aacatattcc attttgtaat ttcgtgtcgt
     5221 ttctattatg aatttcattt ataaagttta tgtacaaata tcataaaaaa agagaatctt
     5281 tttaagcaag gattttctta acttcttcgg cgacagcatc accgacttcg gtggtactgt
     5341 tggaaccacc taaatcacca gttctgatac ctgcatccaa aaccttttta actgcatctt
     5401 caatggcctt accttcttca ggcaagttca atgacaattt caacatcatt gcagcagaca
     5461 agatagtggc gatagggttg accttattct ttggcaaatc tggagcagaa ccgtggcatg
     5521 gttcgtacaa accaaatgcg gtgttcttgt ctggcaaaga ggccaaggac gcagatggca
     5581 acaaacccaa ggaacctggg ataacggagg cttcatcgga gatgatatca ccaaacatgt
     5641 tgctggtgat tataatacca tttaggtggg ttgggttctt aactaggatc atggcggcag
     5701 aatcaatcaa ttgatgttga accttcaatg tagggaattc gttcttgatg gtttcctcca
     5761 cagtttttct ccataatctt gaagaggcca aaacattagc tttatccaag gaccaaatag
     5821 gcaatggtgg ctcatgttgt agggccatga aagcggccat tcttgtgatt ctttgcactt
     5881 ctggaacggt gtattgttca ctatcccaag cgacaccatc accatcgtct tcctttctct
     5941 taccaaagta aatacctccc actaattctc tgacaacaac gaagtcagta cctttagcaa
     6001 attgtggctt gattggagat aagtctaaaa gagagtcgga tgcaaagtta catggtctta
     6061 agttggcgta caattgaagt tctttacgga tttttagtaa accttgttca ggtctaacac
     6121 taccggtacc ccatttagga ccacccacag cacctaacaa aacggcatca gccttcttgg
     6181 aggcttccag cgcctcatct ggaagtggaa cacctgtagc atcgatagca gcaccaccaa
     6241 ttaaatgatt ttcgaaatcg aacttgacat tggaacgaac atcagaaata gctttaagaa
     6301 ccttaatggc ttcggctgtg atttcttgac caacgtggtc acctggcaaa acgacgatct
     6361 tcttaggggc agacattaga atggtatatc cttgaaatat atatatatat attgctgaaa
     6421 tgtaaaaggt aagaaaagtt agaaagtaag acgattgcta accacctatt ggaaaaaaca
     6481 ataggtcctt aaataatatt gtcaacttca agtattgtga tgcaagcatt tagtcatgaa
     6541 cgcttctcta ttctatatga aaagccggtt ccggcgctct cacctttcct ttttctccca
     6601 atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca aaaaatttcc
     6661 agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat gttgaggaaa
     6721 aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga gtattcccac
     6781 agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg gccaaacaac
     6841 caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt ttgaacacac
     6901 atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg atgtaattgt
     6961 tgggattcca tttttaataa ggcaataata ttaggtatgt agatatacta gaagttctcc
     7021 tcgaccggt
//
