LOCUS       sequence-43347-e        7685 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 7685)
  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 7685)
  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: pDS191
FEATURES             Location/Qualifiers
     source          1..7685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..399
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     403..419
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     403..419
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(453..469)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(454..470)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      1067..1314
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1083..1101)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    complement(1327..1360)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            complement(1418..2774)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(1418..1615)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(1621..2430)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2344..2363
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2431..2774)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     protein_bind    complement(2829..2862)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2898..2917)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2899..2917)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2927..2944)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2927..2943)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2936..2958)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3088..3543
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3175..3194)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3385..3406
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3674..4477)
                     /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(4478..4698)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4724..4743)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4843..4865
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4903..4921)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4963..5466
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5503..5607
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5608..5676,5677..6468)
                     /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(5826..5845)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6639..7227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7128..7147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7381..7398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(7551..7556,7557..7574,7575..7581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7589..7605
                     /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     7594..7616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7613..7629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7613..7629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7648..7668
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7650..7668
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 catagcttca aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc
       61 atcgccgtac cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct
      121 ctagggtgtc gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt
      181 ttctttttct tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa
      241 tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg
      301 gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt
      361 cttgctcatt agaaagaaag catagcaatc taatctaagt tttctagaac tagtggatcc
      421 cccgggctgc aggaattgga tatcaagctt atcgataccg tcgacctcga caatgccaga
      481 acttggattt agcatatcag gtggtgtcgg gggtagagga aacccattca gacctgatga
      541 tgatggtata tttgtaacaa gggtacaacc tgaaggacca gcatcaaaat tactgcagcc
      601 aggtgataaa attattcagg ctaatggcta cagttttata aatattgaac atggacaagc
      661 agtgtccttg ctaaaaactt tccagaatac agttgaactc atcattgtac gagaagttgg
      721 taacggtgca aaacaagaga ttcgagtgag ggttgaaaaa gacggtggtt ctggaggcgt
      781 ttcttctgtt ccgaccaatc tggaagttgt tgctgcgacc ccgactagcc tgctgatcag
      841 ctgggatgcg tactacgact ctcatgtttc ttattaccgt atcacgtacg gtgaaaccgg
      901 tggtaactcc ccggttcagg aattcactgt acctggttcc aagtctactg ctaccatcag
      961 cggcctgaaa ccgggtgttg actataccat cactgtatac gcacattaca actaccatta
     1021 ctactctagc ccaatctcga ttaactaccg tacctctaga ctcgagtcat gtaattagtt
     1081 atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt
     1141 agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg
     1201 ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt
     1261 atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg
     1321 tacctaataa cttcgtatag catacattat acgaagttat attaagggtt ctcgacgttt
     1381 tcgacactgg atggcggcgt tagtatcgaa tcgacagcag tatagcgacc agcattcaca
     1441 tacgattgac gcatgatatt actttctgcg cacttaactt cgcatctggg cagatgatgt
     1501 cgaggcgaaa aaaaatataa atcacgctaa catttgatta aaatagaaca actacaatat
     1561 aaaaaaacta tacaaatgac aagttcttga aaacaagaat ctttttattg tcagtactga
     1621 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     1681 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     1741 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     1801 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     1861 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
     1921 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     1981 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     2041 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     2101 ttcttctaat acctggaatg ctgttttgcc ggggatcgca gtggtgagta accatgcatc
     2161 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     2221 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     2281 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     2341 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     2401 cctcgaaacg tgagtctttt ccttacccat ggttgtttat gttcggatgt gatgtgagaa
     2461 ctgtatccta gcaagatttt aaaaggaagt atatgaaaga agaacctcag tggcaaatcc
     2521 taacctttta tatttctcta caggggcgcg gcgtggggac aattcaacgc gtctgtgagg
     2581 ggagcgtttc cctgctcgca ggtctgcagc gaggagccgt aatttttgct tcgcgccgtg
     2641 cggccatcaa aatgtatgga tgcaaatgat tatacatggg gatgtatggg ctaaatgtac
     2701 gggcgacagt cacatcatgc ccctgagctg cgcacgtcaa gactgtcaag gagggtattc
     2761 tgggcctcca tgtcgctggc cgggtgaccc ggcggggacg aggcaagcta aacagatctc
     2821 tagacctaat aacttcgtat agcatacatt atacgaagtt atattaaggg ttgtcttaat
     2881 taagggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     2941 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     3001 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3061 tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg
     3121 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3181 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3241 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3301 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3361 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3421 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3481 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     3541 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggta
     3601 ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt
     3661 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt
     3721 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa
     3781 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca
     3841 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt
     3901 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct
     3961 tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca
     4021 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca
     4081 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
     4141 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt
     4201 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
     4261 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg
     4321 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
     4381 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
     4441 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg
     4501 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac
     4561 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg
     4621 agattaccga atcaaaaaaa tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa
     4681 atgatgaatt gaattgaaaa gctgtggtat ggtgcactct cagtacaatc tgctctgatg
     4741 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4801 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4861 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4921 ttttataggt taatgtcatg ataataatgg tttcttagga cggatcgctt gcctgtaact
     4981 tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct gtgtttattt
     5041 atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa gagttacgga
     5101 atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt actttacata
     5161 tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa cgataagtaa
     5221 aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg aaacaattcg
     5281 gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg gcgatccccc
     5341 tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc tttttttact
     5401 ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat ttttatagca
     5461 cgtgatgaaa aggacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5521 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5581 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5641 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5701 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5761 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5821 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5881 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5941 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6001 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6061 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6121 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6181 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6241 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6301 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6361 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6421 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6481 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6541 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6601 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6661 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6721 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6781 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6841 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6901 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6961 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7021 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7081 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7141 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7201 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7261 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7321 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7381 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     7441 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     7501 gagcgcaacg caattaatgt gagttacctc actcattagg caccccaggc tttacacttt
     7561 atgcttccgg ctcctatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     7621 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
     7681 gagct
//
