LOCUS       BBF10K_000474           3182 bp    DNA     circular UNK 09-JUN-2018
DEFINITION  Yeast marker cassette URA3 in pOpen-v3.
ACCESSION   BBF10K_000474
VERSION     BBF10K_000474.1
KEYWORDS    BBFD_0017; Open Yeast Collection Part; OpenMTA; FreeGenes; Golden
            Gate Assembly.
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
            .
REFERENCE   1  (bases 1 to 2001)
  AUTHORS   Keoni Gandall
  TITLE     Direct Submission
REFERENCE   2  (bases 2002 to 3182)
  AUTHORS   Scott Pownall, Open Science Network
  TITLE     Direct Submission
  JOURNAL   https://biobricks.org/freegenes/
FEATURES             Location/Qualifiers
     source          1..2001
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
                     /original_record="pOpen_v3.1"
     primer_bind     complement(24..42)
                     /label="T7 Reverse"
                     /note="/ApEinfo_fwdcolor=#b7e6d7 /ApEinfo_revcolor=#b7e6d7"
                     /original_record="pOpen_v3.1"
     terminator      38..85
                     /label="T7 terminator"
                     /note="/ApEinfo_fwdcolor=#d6b295 /ApEinfo_revcolor=#d6b295"
                     /original_record="pOpen_v3.1"
     primer_bind     complement(86..106)
                     /label="M13 rev"
                     /note="/ApEinfo_fwdcolor=#ff9ccd /ApEinfo_revcolor=#ff9ccd"
                     /original_record="pOpen_v3.1"
     misc_feature    107..121
                     /label="AarI"
                     /note="/ApEinfo_fwdcolor=#b1ff67 /ApEinfo_revcolor=#b1ff67"
                     /original_record="pOpen_v3.1"
     misc_feature    122..172
                     /label="spacer"
                     /note="/ApEinfo_fwdcolor=#b1ff67 /ApEinfo_revcolor=#b1ff67"
                     /original_record="pOpen_v3.1"
     rep_origin      complement(173..761)
                     /direction=LEFT
                     /label="ori"
                     /note="/ApEinfo_fwdcolor=#84b0dc /ApEinfo_revcolor=#84b0dc"
                     /original_record="pOpen_v3.1"
     CDS             complement(952..1812)
                     /label="AmpR"
                     /note="/ApEinfo_fwdcolor=#ff9ccd /ApEinfo_revcolor=#ff9ccd"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW*"
                     /original_record="pOpen_v3.1"
     misc_feature    1918..1932
                     /label="AarI"
                     /note="/ApEinfo_fwdcolor=#d6b295 /ApEinfo_revcolor=#d6b295"
                     /original_record="pOpen_v3.1"
     primer_bind     1933..1949
                     /label="M13 fwd"
                     /note="/ApEinfo_fwdcolor=#84b0dc /ApEinfo_revcolor=#84b0dc"
                     /original_record="pOpen_v3.1"
     terminator      1950..1981
                     /label="tonB terminator"
                     /note="/ApEinfo_fwdcolor=#b4abac /ApEinfo_revcolor=#b4abac"
                     /original_record="pOpen_v3.1"
     source          2022..3102
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
                     /original_record="part"
     gene            2033..3091
                     /label="URA3 marker"
                     /note="URA3 encodes orotidine-5’phosphate decarboxylase, an
                     enzyme which is required for the biosynthesis of uracil.
                     Complements Ura3 auxotrophic mutations in yeast. It also
                     acts as negative selection is based on the inhibitor
                     5-fluoro-orotic acid (FOA)"
                     /note="/ApEinfo_fwdcolor=#ff9ccd /ApEinfo_revcolor=#ff9ccd"
                     /original_record="part"
     promoter        2033..2209
                     /label="URA3 promoter"
                     /original_record="part"
     CDS             2210..3013
                     /codon_start=1
                     /gene="Ura3"
                     /product="Orotidine-5'-phosphate (OMP) decarboxylase"
                     /label="URA3"
                     /note="Complements URA3 auxotrophic mutations in yeast.
                     URA3 mutants require uracil for growth"
                     /note="/ApEinfo_fwdcolor=#ff9ccd /ApEinfo_revcolor=#ff9ccd"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
                     /original_record="part"
     misc_feature    3014..3091
                     /label="URA3 terminator"
                     /original_record="part"
     protein_bind    complement(join(3092..3095,3097..3102))
                     /label="BsaI(1)"
                     /ApEinfo_label="BsaI"
                     /ApEinfo_fwdcolor="#00FF2C"
                     /ApEinfo_revcolor="#00FF2C"
                     /original_record="part"
     misc_feature    2002..2021
                     /label="FreeGenes dialout primer"
                     /ApEinfo_fwdcolor="#0048BA"
                     /ApEinfo_revcolor="#0048BA"
     misc_feature    complement(3103..3122)
                     /label="FreeGenes dialout primer"
                     /ApEinfo_fwdcolor="#0048BA"
                     /ApEinfo_revcolor="#0048BA"
     misc_feature    3123..3182
                     /label="FreeGenes barcode"
                     /ApEinfo_fwdcolor="#FF7E00"
                     /ApEinfo_revcolor="#FF7E00"
     protein_bind    join(2022..2027,2029..2032)
                     /label="BsaI"
                     /ApEinfo_fwdcolor="#00FF2C"
                     /ApEinfo_revcolor="#00FF2C"
     protein_bind    complement(join(3092..3095,3097..3102))
                     /label="BsaI"
                     /ApEinfo_fwdcolor="#00FF2C"
                     /ApEinfo_revcolor="#00FF2C"
     protein_bind    3144..3161
                     /label="I-SceI"
                     /ApEinfo_fwdcolor="#4F0080"
                     /ApEinfo_revcolor="#4F0080"
ORIGIN
        1 cgctgaggtg tcaatcgtcg gagccgctga gcaataacta gcataacccc ttggggcctc
       61 taaacgggtc ttgaggggtt ttttgcatgg tcatagctgt ttcctgagag cttggcaggt
      121 gatgacacac attaacaaat ttcgtgagga gtctccagaa gaatgccatt aatttccata
      181 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      241 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      301 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      361 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      421 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      481 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
      541 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
      601 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
      661 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
      721 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaaggccta ctattagcaa
      781 caacgatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
      841 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      901 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
      961 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1021 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1081 aatgataccg cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc
     1141 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1201 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     1261 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1321 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     1381 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     1441 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     1501 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     1561 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     1621 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     1681 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     1741 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     1801 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1861 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     1921 ctgcaccagt cagtaaaacg acggccagta gtcaaaagcc tccgaccgga ggcttttgac
     1981 ttggttcagg tggagtggga gaagaattac tgacccctcg gggtctcaaa ggcggtttcc
     2041 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
     2101 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
     2161 cttaacccaa ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc
     2221 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
     2281 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
     2341 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
     2401 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
     2461 caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca
     2521 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
     2581 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
     2641 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
     2701 tactaagggt actgttgaca ttgctaagag cgacaaagat tttgttatcg gctttattgc
     2761 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     2821 gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt
     2881 ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     2941 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     3001 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     3061 agcttcaatt taattatatc agttattacc catgatgaga ccacagaacg aacaggcact
     3121 acacagtctt attttcatcg tcttagggat aacagggtaa tcactaatca tgccttgcgc
     3181 gc
//
