LOCUS BBF10K_000238 3921 bp DNA circular UNK 16-JAN-2020 DEFINITION Yeast cds 4S-limonene-synthase in pOpen-v3. ACCESSION BBF10K_000238 VERSION BBF10K_000238.1 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 3921) 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" CDS 2030..3829 /codon_start=1 /gene="4S-limonene synthase" /product="4S-limonene synthase" /label="4S-limonene synthase" /translation="MALKVLSVATQMAIPSNLTTCLQPSHFKSSPKLLSSTNSSSRSRL RVYCSSSQLTTERRSGNYNPSRWDVNFIQSLLSDYKEDKHVIRASELVTLVKMELEKET DQIRQLELIDDLQRMGLSDHFQNEFKEILSSIYLDHHYYKNPFPKEERDLYSTSLAFRL LREHGFQVAQEVFDSFKNEEGEFKESLSDDTRGLLQLYEASFLLTEGETTLESAREFAT KFLEEKVNEGGVDGDLLTRIAYSLDIPLHWRIKRPNAPVWIEWYRKRPDMNPVVLELAI LDLNIVQAQFQEELKESFRWWRNTGFVEKLPFARDRLVECYFWNTGIIEPRQHASARIM MGKVNALITVIDDIYDVYGTLEELEQFTDLIRRWDINSIDQLPDYMQLCFLALNNFVDD TSYDVMKEKGVNVIPYLRQSWVDLADKYMVEARWFYGGHKPSLEEYLENSWQSISGPCM LTHIFFRVTDSFTKETVDSLYKYHDLVRWSSFVLRLADDLGTSVEEVSRGDVPKSLQCY MSDYNASEAEARKHVKWLIAEVWKKMNAERVSKDSPFGKDFIGCAVDLGRMAQLMYHNG DGHGTQHPIIHQQMTRTLFEPFA" /original_record="part" protein_bind complement(join(3824..3826,3828..3834)) /label="SapI" /ApEinfo_fwdcolor="#00FF2C" /ApEinfo_revcolor="#00FF2C" /original_record="part" protein_bind 3835..3840 /label="HindIII" /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(3842..3861) /label="FreeGenes dialout primer" /ApEinfo_fwdcolor="#0048BA" /ApEinfo_revcolor="#0048BA" misc_feature 3862..3921 /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(3837..3840,3842..3847)) /label="BsaI" /ApEinfo_fwdcolor="#00FF2C" /ApEinfo_revcolor="#00FF2C" protein_bind 3889..3906 /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 gaacgatgat gctcactctc gggtctctaa tggctttaaa 2041 agttttgtct gttgctacac aaatggctat accatcaaat cttactactt gtttgcaacc 2101 atctcatttt aaatcttctc caaagctatt gtctagtact aactcttcta gtagatcgag 2161 attgagagtt tattgttcat catctcaatt aactactgaa aggagatcgg gtaattataa 2221 tccatctagg tgggacgtta attttatcca atctttattg agtgactata aagaggataa 2281 gcatgttatt agagctagtg agttggttac tttggttaag atggaattag aaaaagaaac 2341 tgaccaaatt agacagttgg aactaatcga tgacttgcaa cgtatgggtt tgtcagatca 2401 ttttcaaaat gaatttaagg aaatcttgtc atctatttac ttagaccatc attattataa 2461 gaatccattc ccaaaggaag aaagggactt gtattctacg tctttggcat ttcgcttgtt 2521 gagagaacat ggttttcaag ttgctcaaga agtctttgac tcttttaaga acgaagaagg 2581 cgaatttaag gagtctttat ctgacgatac tagaggtttg ttgcaattat atgaagctag 2641 tttcttgtta actgaaggtg aaactacttt agaatctgct agggaatttg ctactaaatt 2701 tttagaagaa aaggtaaacg aaggtggtgt cgacggtgat ttgcttacta ggatagctta 2761 ttctttagac attccattgc actggagaat taaaggtcca aatgctccag tctggattga 2821 atggtacaga aagaggccag atatgaatcc agttgttttg gagttggcta ttttggactt 2881 gaatatagtt caagcacaat tccaagaaga attgaaagaa tcttttcgat ggtggagaaa 2941 tactggtttt gttgaaaaat tgccttttgc tagagacaga ttggttgagt gttatttctg 3001 gaacactggt attattgaac caagacaaca cgctagtgct agaattatga tgggtaaagt 3061 taatgctttg attactgtta tagacgatat ttacgatgtt tatggtacct tggaggaatt 3121 ggaacaattc actgacttga taagacgttg ggatattaac tctattgacc aattgccaga 3181 ttacatgcaa ttgtgttttt tggctttgaa taatttcgtt gatgatactt cttatgacgt 3241 tatgaaggaa aaaggtgtta atgttatccc atatttgaga caatcttggg ttgacctagc 3301 tgataaatat atggttgaag ctcgttggtt ttacggtggt cacaagccat ctctcgaaga 3361 atatttggaa aattcttggc aatctatctc tggtccatgt atgttgacac atattttttt 3421 tagggttact gattcattca ctaaggaaac tgttgattcc ttgtataaat accatgattt 3481 agttagatgg tcatctttcg ttttgagatt ggctgatgat ttgggtactt ctgttgaaga 3541 agtttcaaga ggtgacgttc caaagtcttt gcaatgttac atgtctgatt acaatgcttc 3601 tgaagcagaa gctagaaaac atgttaagtg gttgattgct gaagtttgga agaagatgaa 3661 tgctgaaaga gtttctaagg atagtccatt tggtaaggat ttcataggtt gtgctgttga 3721 cttgggtaga atggctcaat tgatgtatca caatggtgac ggtcacggta ctcaacaccc 3781 aattatccac caacaaatga ctagaacttt gttcgaacca tttgcttgaa gagcaagctt 3841 tgagaccagc tcaatctaac agtggggttt agaggtccag acggggccta gggataacag 3901 ggtaatccca gaccgcgcaa c //