LOCUS       Exported                5027 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pDS296
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5027)
  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 5027)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5027
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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      1150..1397
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1166..1184)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1472..1491)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1591..1613
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1731..2534)
                     /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(2535..2750)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(2762..2780)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2848..2952
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2953..3813
                     /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(3171..3190)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3984..4572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4473..4492
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4726..4743
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4860..4881
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4896..4926
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4934..4950
                     /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     4939..4961
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4958..4974
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4958..4974
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4992..5010
                     /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 aaagacggtg gttctggagg cgtttcttct gttccgacca atctggaagt tgttgctgcg
      901 accccgacta gcctgctgat cagctgggat gcgtataggg agcttccggt tagttattac
      961 cgtatcacgt acggtgaaac cggtggtaac tccccggttc aggaattcac tgtacctggt
     1021 tccaagtcta ctgctaccat cagcggcctg aaaccgggtg ttgactatac catcactgta
     1081 tacgcacatt acaactacca ttactactct agcccaatct cgattaacta ccgtacctct
     1141 agactcgagt catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg
     1201 ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta
     1261 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     1321 gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct
     1381 cgaaggcttt aatttgcggc cggcgcctga tgcggtattt tctccttacg catctgtgcg
     1441 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     1501 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     1561 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     1621 cgtcatcacc gaaacgcgcg agacgaaagg gcgggtaata actgatataa ttaaattgaa
     1681 gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt ttagttttgc
     1741 tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc accctctacc
     1801 ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc agatcctgta
     1861 gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt gtcatctaaa
     1921 cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc catgtctctt
     1981 tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac agtaccctta
     2041 gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat caaaaggcct
     2101 ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat acctgggccc
     2161 accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac acccgcagag
     2221 tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa
     2281 ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga aaaatcagtc
     2341 aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc aactaactcc
     2401 agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg cttttcgtgc
     2461 atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg ttccttatat
     2521 gtagctttcg acatgattta tcttcgtttc ctgcatgttt ttgttctgtg cagttgggtt
     2581 aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata taccaatcta
     2641 agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt
     2701 caaagaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa ttgaaaagct
     2761 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     2821 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2881 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2941 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3001 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3061 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3121 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3181 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3241 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3301 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3361 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3421 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3481 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3541 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3601 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3661 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3721 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3781 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3841 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3901 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3961 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4021 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4081 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     4141 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4201 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4261 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4321 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4381 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4441 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4501 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4561 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4621 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4681 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4741 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4801 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4861 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     4921 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     4981 tacgccaagc taattaaccc tcactaaagg gaacaaaagc tggagct
//
