LOCUS       sequence-75766-e        6592 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6592)
  AUTHORS   Markus SM, Omer S, Baranowski K, Lee WL
  TITLE     Improved Plasmids for Fluorescent Protein Tagging of Microtubules in
            Saccharomyces cerevisiae.
  JOURNAL   Traffic. 2015 Jul;16(7):773-86. doi: 10.1111/tra.12276. Epub 2015 
            Apr 28.
  PUBMED    25711127
REFERENCE   2  (bases 1 to 6592)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1111/tra.12276";
            journalName: "Traffic"; date: "2015-07"; volume: "16"; issue: "7"; 
            pages: "773-86"
COMMENT     Sequence Label: pHIS3p:mRuby2-Tub1+3'UTR::TRP1
FEATURES             Location/Qualifiers
     source          1..6592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     72..94
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(132..150)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        218..322
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(323..391,392..1183)
                     /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(541..560)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1354..1942
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1843..1862
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2096..2113
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2200..2219
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2200..2218
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2303..2583
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             2584..3258
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        3423..3609
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3611..4321
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     RBS             5806..5814
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(6446..6464)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6447..6464)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
       61 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
      121 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
      181 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      241 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      301 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      361 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      421 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      481 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      541 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
      601 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      661 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      721 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      781 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
      841 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
      901 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
      961 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1021 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1081 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1141 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1201 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1261 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1321 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1381 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1441 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1501 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1561 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1621 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1681 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1741 tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1801 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1861 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1921 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     1981 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2041 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2101 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     2161 gccgattcat taatgcaggt taacctggct tatcgaaatt aatacgactc actataggga
     2221 gaccggcaga tccgcggccg cataggccac tagttaacta tgcggcatca gagcagattg
     2281 tactgagagt gcaccataga tcacgacatt actatatata taatatagga agcatttaat
     2341 agaacagcat cgtaatatat gtgtactttg cagttatgac gccagatggc agtagtggaa
     2401 gatattcttt attgaaaaat agcttgtcac cttacgtaca atcttgatcc ggagcttttc
     2461 tttttttgcc gattaagaat taattcggtc gaaaaaagaa aaggagaggg ccaagaggga
     2521 gggcattggt gactattgag cacgtgagta tacgtgatta agcacacaaa ggcagcttgg
     2581 agtatgtctg ttattaattt cacaggtagt tctggtccat tggtgaaagt ttgcggcttg
     2641 cagagcacag aggccgcaga atgtgctcta gattccgatg ctgacttgct gggtattata
     2701 tgtgtgccca atagaaagag aacaattgac ccggttattg caaggaaaat ttcaagtctt
     2761 gtaaaagcat ataaaaatag ttcaggcact ccgaaatact tggttggcgt gtttcgtaat
     2821 caacctaagg aggatgtttt ggctctggtc aatgattacg gcattgatat cgtccaactg
     2881 catggagatg agtcgtggca agaataccaa gagttcctcg gtttgccagt tattaaaaga
     2941 ctcgtatttc caaaagactg caacatacta ctcagtgcag cttcacagaa acctcattcg
     3001 tttattccct tgtttgattc agaagcaggt gggacaggtg aacttttgga ttggaactcg
     3061 atttctgact gggttggaag gcaagagagc cccgaaagct tacattttat gttagctggt
     3121 ggactgacgc cagaaaatgt tggtgatgcg cttagattaa atggcgttat tggtgttgat
     3181 gtaagcggag gtgtggagac aaatggtgta aaagactcta acaaaatagc aaatttcgtc
     3241 aaaaatgcta agaaataggt tattactgag tagtatttat ttaagtattg tttgtgcact
     3301 tgccgatcta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     3361 ggcgcgcctt aattaaccac taaagggaac aaaagctggt accctattac tcttggcctc
     3421 ctctagtaca ctctatattt ttttatgcct cggtaatgat tttcattttt tttttccacc
     3481 tagcggatga ctcttttttt ttcttagcga ttggcattat cacataatga attatacatt
     3541 atataaagta atgtgatttc ttcgaagaat atactaaaaa atgagcaggc aagataaacg
     3601 aaggcaaagc atggtgtcta agggcgaaga gctgatcaag gaaaatatgc gtatgaaggt
     3661 ggtcatggaa ggttcggtca acggccacca attcaaatgc acaggtgaag gagaaggcaa
     3721 tccgtacatg ggaactcaaa ccatgaggat caaagtcatc gagggaggac ccctgccatt
     3781 tgcctttgac attcttgcca cgtcgttcat gtatggcagc cgtactttta tcaagtaccc
     3841 gaaaggcatt cctgatttct ttaaacagtc ctttcctgag ggttttactt gggaaagagt
     3901 tacgagatac gaagatggtg gagtcgtcac cgtcatgcag gacaccagcc ttgaggatgg
     3961 ctgtctcgtt taccacgtcc aagtcagagg ggtaaacttt ccctccaatg gtcccgtgat
     4021 gcagaagaag accaagggtt gggagcctaa tacagagatg atgtatccag cagatggtgg
     4081 tctgagggga tacactcata tggcactgaa agttgatggt ggtggccatc tgtcttgctc
     4141 tttcgtaaca acttacaggt caaaaaagac cgtcgggaac atcaagatgc ccggtatcca
     4201 tgccgttgat caccgcctgg aaaggttaga ggaaagtgac aatgaaatgt tcgtagtaca
     4261 acgcgaacac gcagttgcca agttcgccgg gcttggtggt gggatggacg agctgtacaa
     4321 gggatccatg agagaagtta ttagtattaa tggtatgttc gatttgcccg tccaggctag
     4381 atcttttttt tgttagttca ttttggcgtt tcatctttca ttactaactt caacgcaaaa
     4441 tttttttttt tttgatttct ctttacagtc ggtcaagctg gttgtcagat tggtaatgcc
     4501 tgttgggaat tatattccct tgagcacggt attaagccgg atggacatct agaagatggc
     4561 ctttcaaagc cgaagggagg agaagagggt ttctccacgt ttttccatga aaccggctac
     4621 ggtaagttcg ttccaagggc tatttacgtg gatttagagc ccaacgttat tgacgaagtc
     4681 cgtaacggtc cttacaagga cttgttccat ccagaacagt tgatcagtgg taaggaggac
     4741 gcggctaata attatgcaag aggccattac accgttggta gagaaatttt gggcgatgtt
     4801 ctggatagga ttagaaaact ggcagaccaa tgtgatgggt tacaagggtt cttgtttacc
     4861 cattctcttg gtggtggtac tggttccggt ctagggtcct tactgttgga agaactatcc
     4921 gctgaatacg gtaagaaatc caagctggaa tttgccgtat accctgctcc acaagtgtct
     4981 acttctgtgg ttgagcctta caacacagtt ttaactacac atactacatt ggaacatgca
     5041 gattgtactt tcatggtcga taatgaggct atttacgaca tgtgcaaaag aaacttggat
     5101 atcccaagac caagctttgc aaacttaaac aacctaattg ctcaagtggt atcatctgtt
     5161 acagcatcat tgagattcga cggttcatta aatgtagatt tgaacgaatt tcaaaccaat
     5221 ttggttccat atccaagaat tcatttcccc ttagtttcat attctccagt cttatccaaa
     5281 tcaaaggcat tccatgagtc caactctgtg tcagaaatta caaacgcttg ttttgaacct
     5341 ggtaaccaga tggtcaagtg tgatccaaga gatggtaaat acatggctac ttgtctgtta
     5401 tacaggggtg atgtggtaac aagagatgtt caaagagctg tcgagcaggt gaaaaacaag
     5461 aagaccgtcc aattggttga ttggtgtcca actggtttca agatcggtat ttgctacgaa
     5521 cctccaactg ccacaccaaa ctcacaattg gccactgtgg atagggccgt ctgtatgttg
     5581 tcaaatacca catccattgc tgaggcttgg aagagaatcg atagaaaatt cgatttaatg
     5641 tatgccaaac gtgctttcgt ccactggtat gtcggtgaag gtatggaaga aggtgaattc
     5701 accgaagcta gagaagattt ggctgcttta gaaagagatt acatcgaagt gggtgccgac
     5761 tcatacgctg aggaagagga attttaaggt tcactctcgc ccccaacctc cttatccttt
     5821 ctatatttca agtatattgt aaaaaaaaaa attcccccta ctgttatatc tctttggtga
     5881 tatcttatct ataatctact tattatattt ctctaatatc tatgtacaaa aaataaaaaa
     5941 aaaaaccatt ctccttctct aacctgcatt tgatgaaagc ttagtaagtc tcatcgtagt
     6001 tgatgggcca aagacatttg tacttgtttc atcagattct tacatcaccc atttccatga
     6061 ttatttcacg gatttcatta gtaattgcat ttttgtatcc ctagttcctt ttctcccgcc
     6121 ttgccgtcga attgaaaaaa aaaatggaaa tgaggacatc ttccattgat atccaggaca
     6181 aagcaagagt agaagtaact attagcaagg agaattccaa aaatagtgaa cttcaaggta
     6241 agcagatcta tagtttcaat atctgatttt ctgcttttca aaacttaaaa ttcattagaa
     6301 ggtaaaacaa gaacccgtaa aattaaagtt accgatatta tcaggttttt caccccatgt
     6361 ctactatccc atttagaaag aactatgtgt tcaaaaactg ggccggtcga cctgcagcgt
     6421 acgaagcttc agctggcggc cgcgttctat agtgtcacct aaatcgtatg tgtatgatac
     6481 ataaggttat gtattaattg tagccgcgtt ctaacgacaa tatgtccata tggtgcactc
     6541 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg cc
//
