LOCUS       sequence-75770-e        6772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6772)
  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 6772)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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:yomWasabi-Tub1+3'UTR::HIS3
FEATURES             Location/Qualifiers
     source          1..6772
                     /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        2429..2616
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             2617..3279
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        3601..3787
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     RBS             5986..5994
                     /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(6626..6644)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6627..6644)
                     /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 gcaccataaa ttcccgtttt aagagcttgg tgagcgctag gagtcactgc
     2341 caggtatcgt ttgaacacgg cattagtcag ggaagtcata acacagtcct ttcccgcaat
     2401 tttctttttc tattactctt ggcctcctct agtacactct atattttttt atgcctcggt
     2461 aatgattttc attttttttt ttccacctag cggatgactc tttttttttc ttagcgattg
     2521 gcattatcac ataatgaatt atacattata taaagtaatg tgatttcttc gaagaatata
     2581 ctaaaaaatg agcaggcaag ataaacgaag gcaaagatga cagagcagaa agccctagta
     2641 aagcgtatta caaatgaaac caagattcag attgcgatct ctttaaaggg tggtccccta
     2701 gcgatagagc actcgatctt cccagaaaaa gaggcagaag cagtagcaga acaggccaca
     2761 caatcgcaag tgattaacgt ccacacaggt atagggtttc tggaccatat gatacatgct
     2821 ctggccaagc attccggctg gtcgctaatc gttgagtgca ttggtgactt acacatagac
     2881 gaccatcaca ccactgaaga ctgcgggatt gctctcggtc aagcttttaa agaggcccta
     2941 ggggccgtgc gtggagtaaa aaggtttgga tcaggatttg cgcctttgga tgaggcactt
     3001 tccagagcgg tggtagatct ttcgaacagg ccgtacgcag ttgtcgaact tggtttgcaa
     3061 agggagaaag taggagatct ctcttgcgag atgatcccgc attttcttga aagctttgca
     3121 gaggctagca gaattaccct ccacgttgat tgtctgcgag gcaagaatga tcatcaccgt
     3181 agtgagagtg cgttcaaggc tcttgcggtt gccataagag aagccacctc gcccaatggt
     3241 accaacgatg ttccctccac caaaggtgtt cttatgtagt gacaccgatt atttaaagct
     3301 gcagcatacg atatatatac atgtgtatat atgtatacct atgaatgtca gtaagtatgt
     3361 atacgaacag tatgatactg aagatgacaa ggtaatgcat cattctatac gtgtcattct
     3421 gaacgaggcg cgctttcctt ttttcttttt gctttttctt tttttttctc ttgaactcga
     3481 cggatctatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcagggg
     3541 cgcgccttaa ttaaccacta aagggaacaa aagctggtac cctattactc ttggcctcct
     3601 ctagtacact ctatattttt ttatgcctcg gtaatgattt tcattttttt tttccaccta
     3661 gcggatgact cttttttttt cttagcgatt ggcattatca cataatgaat tatacattat
     3721 ataaagtaat gtgatttctt cgaagaatat actaaaaaat gagcaggcaa gataaacgaa
     3781 ggcaaagctc gagatggtaa gtaaaggtga agagacaact atgggggtaa tcaaaccaga
     3841 catgaagatc aaactgaaaa tggaaggtaa cgttaatggc cacgcgtttg ttattgaagg
     3901 tgaaggagag gggaaacctt acgacggaac caacactatc aatttggagg ttaaggaagg
     3961 tgcaccatta ccattcagct atgacatctt aacaactgca ttttcatacg gcaacagagc
     4021 ttttaccaag taccctgacg atataccaaa ttacttcaaa caatcctttc ctgaaggtta
     4081 ttcatgggag agaacaatga cgtttgaaga taagggcatt gtcaaggtca aatctgacat
     4141 ttctatggaa gaggattcat tcatctacga gattcacctc aagggcgaaa acttcccacc
     4201 aaatggtcca gtgatgcaaa aggaaactac tggatgggat gcttctacag aaaggatgta
     4261 cgtgagagat ggagtcttaa aaggtgatgt taaaatgaag ttgctacttg aaggtggtgg
     4321 ccatcataga gttgatttca aaacaatcta cagagccaaa aaggctgtaa aacttcctga
     4381 ttatcatttt gtggatcatc gtatagaaat tctaaaccac gacaaagatt acaataaggt
     4441 cactgtttat gaaatagcag ttgccagaaa ttctacagat ggaatggatg aattgtacaa
     4501 aggatccatg agagaagtta ttagtattaa tggtatgttc gatttgcccg tccaggctag
     4561 atcttttttt tgttagttca ttttggcgtt tcatctttca ttactaactt caacgcaaaa
     4621 tttttttttt tttgatttct ctttacagtc ggtcaagctg gttgtcagat tggtaatgcc
     4681 tgttgggaat tatattccct tgagcacggt attaagccgg atggacatct agaagatggc
     4741 ctttcaaagc cgaagggagg agaagagggt ttctccacgt ttttccatga aaccggctac
     4801 ggtaagttcg ttccaagggc tatttacgtg gatttagagc ccaacgttat tgacgaagtc
     4861 cgtaacggtc cttacaagga cttgttccat ccagaacagt tgatcagtgg taaggaggac
     4921 gcggctaata attatgcaag aggccattac accgttggta gagaaatttt gggcgatgtt
     4981 ctggatagga ttagaaaact ggcagaccaa tgtgatgggt tacaagggtt cttgtttacc
     5041 cattctcttg gtggtggtac tggttccggt ctagggtcct tactgttgga agaactatcc
     5101 gctgaatacg gtaagaaatc caagctggaa tttgccgtat accctgctcc acaagtgtct
     5161 acttctgtgg ttgagcctta caacacagtt ttaactacac atactacatt ggaacatgca
     5221 gattgtactt tcatggtcga taatgaggct atttacgaca tgtgcaaaag aaacttggat
     5281 atcccaagac caagctttgc aaacttaaac aacctaattg ctcaagtggt atcatctgtt
     5341 acagcatcat tgagattcga cggttcatta aatgtagatt tgaacgaatt tcaaaccaat
     5401 ttggttccat atccaagaat tcatttcccc ttagtttcat attctccagt cttatccaaa
     5461 tcaaaggcat tccatgagtc caactctgtg tcagaaatta caaacgcttg ttttgaacct
     5521 ggtaaccaga tggtcaagtg tgatccaaga gatggtaaat acatggctac ttgtctgtta
     5581 tacaggggtg atgtggtaac aagagatgtt caaagagctg tcgagcaggt gaaaaacaag
     5641 aagaccgtcc aattggttga ttggtgtcca actggtttca agatcggtat ttgctacgaa
     5701 cctccaactg ccacaccaaa ctcacaattg gccactgtgg atagggccgt ctgtatgttg
     5761 tcaaatacca catccattgc tgaggcttgg aagagaatcg atagaaaatt cgatttaatg
     5821 tatgccaaac gtgctttcgt ccactggtat gtcggtgaag gtatggaaga aggtgaattc
     5881 accgaagcta gagaagattt ggctgcttta gaaagagatt acatcgaagt gggtgccgac
     5941 tcatacgctg aggaagagga attttaaggt tcactctcgc ccccaacctc cttatccttt
     6001 ctatatttca agtatattgt aaaaaaaaaa attcccccta ctgttatatc tctttggtga
     6061 tatcttatct ataatctact tattatattt ctctaatatc tatgtacaaa aaataaaaaa
     6121 aaaaaccatt ctccttctct aacctgcatt tgatgaaagc ttagtaagtc tcatcgtagt
     6181 tgatgggcca aagacatttg tacttgtttc atcagattct tacatcaccc atttccatga
     6241 ttatttcacg gatttcatta gtaattgcat ttttgtatcc ctagttcctt ttctcccgcc
     6301 ttgccgtcga attgaaaaaa aaaatggaaa tgaggacatc ttccattgat atccaggaca
     6361 aagcaagagt agaagtaact attagcaagg agaattccaa aaatagtgaa cttcaaggta
     6421 agcagatcta tagtttcaat atctgatttt ctgcttttca aaacttaaaa ttcattagaa
     6481 ggtaaaacaa gaacccgtaa aattaaagtt accgatatta tcaggttttt caccccatgt
     6541 ctactatccc atttagaaag aactatgtgt tcaaaaactg ggccggtcga cctgcagcgt
     6601 acgaagcttc agctggcggc cgcgttctat agtgtcacct aaatcgtatg tgtatgatac
     6661 ataaggttat gtattaattg tagccgcgtt ctaacgacaa tatgtccata tggtgcactc
     6721 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg cc
//
