LOCUS       BBF10K_000471           4041 bp    DNA     circular UNK 09-JUN-2018
DEFINITION  ScLEU2 marker in pOpen-v3.
ACCESSION   BBF10K_000471
VERSION     BBF10K_000471.1
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 4041)
  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..3961
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
                     /original_record="part"
     gene            2034..3950
                     /label="LEU2 marker"
                     /note="Complements Leu2 auxotrophic mutations in yeast.
                     Leu2 mutants require histidine"
                     /note="/ApEinfo_fwdcolor=#ff9ccd /ApEinfo_revcolor=#ff9ccd"
                     /original_record="part"
     promoter        2034..2681
                     /label="LEU2 promoter"
                     /original_record="part"
     CDS             2682..3776
                     /codon_start=1
                     /gene="Leu2"
                     /product="Beta-isopropylmalate dehydrogenase"
                     /label="LEU2"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
                     /note="/ApEinfo_fwdcolor=#ff9ccd /ApEinfo_revcolor=#ff9ccd"
                     /original_record="part"
     misc_feature    3777..3950
                     /label="LEU2 terminator"
                     /original_record="part"
     protein_bind    complement(join(3951..3954,3956..3961))
                     /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(3962..3981)
                     /label="FreeGenes dialout primer"
                     /ApEinfo_fwdcolor="#0048BA"
                     /ApEinfo_revcolor="#0048BA"
     misc_feature    3982..4041
                     /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(3951..3954,3956..3961))
                     /label="BsaI"
                     /ApEinfo_fwdcolor="#00FF2C"
                     /ApEinfo_revcolor="#00FF2C"
     protein_bind    4003..4020
                     /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 ggatcgagga
     2041 gaacttctag tatatctaca tacctaatat tattgcctta ttaaaaatgg aatcccaaca
     2101 attacatcaa aatccacatt ctcttcaaaa tcaattgtcc tgtacttcct tgttcatgtg
     2161 tgttcaaaaa cgttatattt ataggataat tatactctat ttctcaacaa gtaattggtt
     2221 gtttggccga gcggtctaag gcgcctgatt caagaaatat cttgaccgca gttaactgtg
     2281 ggaatactca ggtatcgtaa gatgcaagag ttcgaatctc ttagcaacca ttattttttt
     2341 cctcaacata acgagaacac acaggggcgc tatcgcacag aatcaaattc gatgactgga
     2401 aattttttgt taatttcaga ggtcgcctga cgcatatacc tttttcaact gaaaaattgg
     2461 gagaaaaagg aaaggtgaga gcgccggaac cggcttttca tatagaatag agaagcgttc
     2521 atgactaaat gcttgcatca caatacttga agttgacaat attatttaag gacctattgt
     2581 tttttccaat aggtggttag caatcgtctt actttctaac ttttcttacc ttttacattt
     2641 cagcaatata tatatatata tttcaaggat ataccattct aatgtctgcc cctaagaaga
     2701 tcgtcgtttt gccaggtgac cacgttggtc aagaaatcac agccgaagcc attaaggttc
     2761 ttaaagctat ttctgatgtt cgttccaatg tcaagttcga tttcgaaaat catttaattg
     2821 gtggtgctgc tatcgatgct acaggtgttc cacttccaga tgaggcgctg gaagcctcca
     2881 agaaggctga tgccgttttg ttaggtgctg tgggtggtcc taaatgggga accggtagtg
     2941 ttagacctga acaaggttta ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa
     3001 gaccatgtaa ctttgcatcc gactctcttt tagacttatc tccaatcaag ccacaatttg
     3061 ctaaaggtac tgacttcgtt gttgtcagag aattagtggg aggtatttac tttggtaaga
     3121 gaaaggagga cgatggtgat ggtgtcgctt gggatagtga acaatacacc gttccagaag
     3181 tgcaaagaat cacaagaatg gccgctttca tggccctaca acatgagcca ccattgccta
     3241 tttggtcctt ggataaagct aatgttttgg cctcttcaag attatggaga aaaactgtgg
     3301 aggaaaccat caagaacgaa tttcctacat tgaaggttca acatcaattg attgattctg
     3361 ccgccatgat cctagttaag aacccaaccc acctaaatgg tattataatc accagcaaca
     3421 tgtttggtga tatcatctcc gatgaagcct ccgttatccc aggttccttg ggtttgttgc
     3481 catctgcgtc cttggcctct ttgccagaca agaacaccgc atttggtttg tacgaaccat
     3541 gccacggttc tgctccagat ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt
     3601 ctgctgcaat gatgttgaaa ttgtcattga acttgcctga agaaggtaag gccattgaag
     3661 atgcagttaa aaaggttttg gatgcaggta tcagaactgg tgatttaggt ggttccaaca
     3721 gtaccaccga agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt gcttaacctg
     3781 gaggaccctt ctctttagac tattctactc ttatgcacgt aaaaaattct aggaaatatg
     3841 tattaactag gagtaaaata accggctagt ggcattcata tagccgtctg tttacatcta
     3901 catcacacat ttcgagtgta tatctcgcaa cgttggcgtt aaataggcag atgatgagac
     3961 caatgtcctt aggcagtcgt ctggatgtat tcagaacggc gctagggata acagggtaat
     4021 tttgtaaaaa ttaaatttcg g
//
