LOCUS Exported 5600 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDS282 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5600 /organism="synthetic DNA construct" /mol_type="other DNA" 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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(1910..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(3023..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 4086..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 //