LOCUS       sequence-43363-e        4736 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 4736)
  AUTHORS   Strickland D, Lin Y, Wagner E, Hope CM, Zayner J, Antoniou C, 
            Sosnick TR, Weiss EL, Glotzer M
  TITLE     TULIPs: tunable, light-controlled interacting protein tags for cell 
            biology.
  JOURNAL   Nat Methods. 2012 Mar 4. doi: 10.1038/nmeth.1904.
  PUBMED    22388287
REFERENCE   2  (bases 1 to 4736)
  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: "Nat 
            Methods. 2012 Mar 4. doi: 10.1038/nmeth.1904."
COMMENT     Sequence Label: pDS297
FEATURES             Location/Qualifiers
     source          1..4736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..464
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    23..140
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     435..458
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(536..552)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(537..553)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      859..1106
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(875..893)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1181..1200)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1300..1322
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1440..2243)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(2244..2464)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(2471..2489)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2557..2661
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2662..2730,2731..3522)
                     /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(2880..2899)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3693..4281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4182..4201
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4435..4452
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4569..4590
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4605..4610,4611..4628,4629..4635)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4643..4659
                     /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     4648..4670
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4667..4683
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4667..4683
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4701..4719
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cgtttaaacg gcgcgcctag tacggattag aagccgccga gcgggtgaca gccctccgaa
       61 ggaagactct cctccgtgcg tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg
      121 cctcgcgccg cactgctccg aacaataaag attctacaat actagctttt atggttatga
      181 agaggaaaaa ttggcagtaa cctggcccca caaaccttca aatgaacgaa tcaaattaac
      241 aaccatagga tgataatgcg attagttttt tagccttatt tctggggtaa ttaatcagcg
      301 aagcgatgat ttttgatcta ttaacagata tataaatgca aaaactgcat aaccacttta
      361 actaatactt tcaacatttt cggtttgtat tacttcttat tcaaatgtaa taaaagtatc
      421 aacaaaaaat tgttaatata cctctatact ttaacgtcaa ggagaaaaac cccggattct
      481 gaagcggccg cactagtgga tcccccgggc tgcaggaatt ggatatcaag cttatcgata
      541 ccgtcgacct cgacaatgcc agaacttgga tttagcatat caggtggtgt cgggggtaga
      601 ggaaacccat tcagacctga tgatgatggt atatttgtaa caagggtaca acctgaagga
      661 ccagcatcaa aattactgca gccaggtgat aaaattattc aggctaatgg ctacagtttt
      721 ataaatattg aacatggaca agcagtgtcc ttgctaaaaa ctttccagaa tacagttgaa
      781 ctcatcattg tacgagaagt tggtaacggt gcaaaacaag agattcgagt gagggttgaa
      841 aaagactcta gactcgagtc atgtaattag ttatgtcacg cttacattca cgccctcccc
      901 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
      961 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     1021 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
     1081 tgggacgctc gaaggcttta atttgcggcc ggcgcctgat gcggtatttt ctccttacgc
     1141 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     1201 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     1261 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1321 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cgggtaataa ctgatataat
     1381 taaattgaag ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt
     1441 tagttttgct ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca
     1501 ccctctacct tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca
     1561 gatcctgtag agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg
     1621 tcatctaaac ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc
     1681 atgtctcttt gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca
     1741 gtacccttag tatattctcc agtagatagg gagcccttgc atgacaattc tgctaacatc
     1801 aaaaggcctc taggttcctt tgttacttct tctgccgcct gcttcaaacc gctaacaata
     1861 cctgggccca ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca
     1921 cccgcagagt actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag
     1981 agtaaaaaat tgtacttggc ggataatgcc tttagcggct taactgtgcc ctccatggaa
     2041 aaatcagtca agatatccac atgtgttttt agtaaacaaa ttttgggacc taatgcttca
     2101 actaactcca gtaattcctt ggtggtacga acatccaatg aagcacacaa gtttgtttgc
     2161 ttttcgtgca tgatattaaa tagcttggca gcaacaggac taggatgagt agcagcacgt
     2221 tccttatatg tagctttcga catgatttat cttcgtttcc tgcatgtttt tgttctgtgc
     2281 agttgggtta agaatactgg gcaatttcat gtttcttcaa cactacatat gcgtatatat
     2341 accaatctaa gtctgtgctc cttccttcgt tcttccttct gttcggagat taccgaatca
     2401 aaaaaatttc aaagaaaccg aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaat
     2461 tgaaaagctg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     2521 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2581 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2641 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2701 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2761 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2821 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2881 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     2941 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3001 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3061 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3121 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3181 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3241 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3301 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3361 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3421 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3481 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3541 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3601 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3661 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3721 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3781 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3841 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3901 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3961 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4021 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4081 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4141 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4201 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4261 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4321 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4381 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4441 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4501 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4561 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     4621 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     4681 gaccatgatt acgccaagct aattaaccct cactaaaggg aacaaaagct ggagct
//
