LOCUS       sequence-43353-e        5600 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 5600)
  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 5600)
  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: pDS282
FEATURES             Location/Qualifiers
     source          1..5600
                     /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"
     misc_feature    764..816
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      824..833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             830..1540
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(975..993)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1243..1262
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1553..1800
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1569..1587)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1862..1878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1862..1878)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1875..1897)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1886..1902
                     /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(1910..1916,1917..1934,1935..1940))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1955..1976
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2093..2110)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2264..2852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2344..2363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3023..3814,3815..3883))
                     /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     3646..3665
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3884..3988)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4056..4074
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4081..4301
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4302..5105
                     /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"
     primer_bind     complement(5223..5245)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5345..5364
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5558..5580
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5572..5589
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5573..5589
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 aggaattcga tatcaagctt atcgataccg tcgacaatgc cagaacttgg
      481 atttagcata tcaggtggtg tcgggggtag aggaaaccca ttcagacctg atgatgatgg
      541 tatatttgta acaagggtac aacctgaagg accagcatca aaattactgc agccaggtga
      601 taaaattatt caggctaatg gctacagttt tataaatatt gaacatggac aagcagtgtc
      661 cttgctaaaa actttccaga atacagttga actcatcatt gtacgagaag ttggtaacgg
      721 tgcaaaacaa gagattcgag tgagggttga aaaagactct agactcgagc tcaagcttcg
      781 aattctgcaa tcgacggtac cgcgggcccg ggatccaccg gtcgccacca tggtgagcaa
      841 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
      901 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
      961 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
     1021 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     1081 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     1141 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     1201 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     1261 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     1321 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     1381 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     1441 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     1501 ccgccactcc accggcggca tggacgagct gtacaagtaa gtcgacctcg agtcatgtaa
     1561 ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa
     1621 ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta
     1681 agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt
     1741 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc
     1801 ggccggtacc cggggatcct ctagagtcga cctgcaggca tgcaagcttg gcgtaatcat
     1861 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1921 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1981 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2041 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2101 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2161 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2221 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2281 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2341 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2401 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2461 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2521 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2581 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2641 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2701 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2761 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2821 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2881 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2941 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3001 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3061 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3121 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3181 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3241 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3301 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3361 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3421 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3481 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3541 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3601 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3661 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3721 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3781 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3841 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3901 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3961 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4021 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccagctt
     4081 ttcaattcaa ttcatcattt tttttttatt cttttttttg atttcggttt ctttgaaatt
     4141 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     4201 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     4261 aactgcacag aacaaaaaca tgcaggaaac gaagataaat catgtcgaaa gctacatata
     4321 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     4381 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     4441 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     4501 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     4561 tactcttcga agacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     4621 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     4681 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     4741 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     4801 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     4861 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     4921 atgacaaggg agacgcattg ggtcaacagt atagaaccgt ggatgatgtg gtctctacag
     4981 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     5041 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     5101 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     5161 tttaattata tcagttatta cccgcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     5221 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     5281 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     5341 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     5401 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     5461 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     5521 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     5581 acggccagtg aattcgagct
//
