LOCUS       sequence-75755-e        6659 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 6659)
  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 6659)
  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:mEos2-Tub1+3'UTR::URA3
FEATURES             Location/Qualifiers
     source          1..6659
                     /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        2306..2521
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             2522..3325
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        3518..3704
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3711..4388
                     /codon_start=1
                     /product="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
                     /label=mEos2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     RBS             5873..5881
                     /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(6513..6531)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6514..6531)
                     /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 gcaccatacc accttttcaa ttcatcattt tttttttatt cttttttttg
     2341 atttcggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg aagaacgaag
     2401 gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag aaacatgaaa
     2461 ttgcccagta ttcttaaccc aactgcacag aacaaaaacc tgcaggaaac gaagataaat
     2521 catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa
     2581 gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac
     2641 caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac
     2701 acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt
     2761 atccgccaag tacaattttt tactcttcga agacagaaaa tttgctgaca ttggtaatac
     2821 agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa
     2881 tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt
     2941 aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctatc
     3001 tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat
     3061 cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat
     3121 gacacccggt gtgggtttag atgacaaggg agacgcattg ggtcaacagt atagaaccgt
     3181 ggatgatgtg gtctctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa
     3241 gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt
     3301 gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa
     3361 ctcacaaatt agagcttcaa tttaattata tcagttatta ccctatgcgg tgtgaaatac
     3421 cgcacagatg cgtaaggaga aaataccgca tcaggggcgc gccttaatta accactaaag
     3481 ggaacaaaag ctggtaccct attactcttg gcctcctcta gtacactcta tattttttta
     3541 tgcctcggta atgattttca tttttttttt ccacctagcg gatgactctt tttttttctt
     3601 agcgattggc attatcacat aatgaattat acattatata aagtaatgtg atttcttcga
     3661 agaatatact aaaaaatgag caggcaagat aaacgaaggc aaagctcgag atgagtgcga
     3721 ttaagccaga catgaagatc aaactccgta tggaaggcaa cgtaaacggg caccactttg
     3781 tgatcgacgg agatggtaca ggcaagcctt ttgagggaaa acagagtatg gatcttgaag
     3841 tcaaagaggg cggacctctg ccttttgcct ttgatatcct gaccactgca ttccattacg
     3901 gcaacagggt attcgccaaa tatccagaca acatacaaga ctattttaag cagtcgtttc
     3961 ctaaggggta ttcgtgggaa cgaagcttga ctttcgaaga cgggggcatt tgcattgcca
     4021 gaaacgacat aacaatggaa ggggacactt tctataataa agttcgattt tatggtacca
     4081 actttcccgc caatggtcca gttatgcaga agaagacgct gaaatgggag ccctccactg
     4141 agaaaatgta tgtgcgtgat ggagtgctga cgggtgatat tcatatggct ttgttgcttg
     4201 aaggaaatgc ccattaccga tgtgacttca gaactactta caaagctaag gagaagggtg
     4261 tcaagttacc aggctaccac tttgtggacc actgcattga gattttaagc catgacaaag
     4321 attacaacaa ggttaagctg tatgagcatg ctgttgctca ttctggattg cctgacaatg
     4381 ccagacgagg atccatgaga gaagttatta gtattaatgg tatgttcgat ttgcccgtcc
     4441 aggctagatc ttttttttgt tagttcattt tggcgtttca tctttcatta ctaacttcaa
     4501 cgcaaaattt tttttttttt gatttctctt tacagtcggt caagctggtt gtcagattgg
     4561 taatgcctgt tgggaattat attcccttga gcacggtatt aagccggatg gacatctaga
     4621 agatggcctt tcaaagccga agggaggaga agagggtttc tccacgtttt tccatgaaac
     4681 cggctacggt aagttcgttc caagggctat ttacgtggat ttagagccca acgttattga
     4741 cgaagtccgt aacggtcctt acaaggactt gttccatcca gaacagttga tcagtggtaa
     4801 ggaggacgcg gctaataatt atgcaagagg ccattacacc gttggtagag aaattttggg
     4861 cgatgttctg gataggatta gaaaactggc agaccaatgt gatgggttac aagggttctt
     4921 gtttacccat tctcttggtg gtggtactgg ttccggtcta gggtccttac tgttggaaga
     4981 actatccgct gaatacggta agaaatccaa gctggaattt gccgtatacc ctgctccaca
     5041 agtgtctact tctgtggttg agccttacaa cacagtttta actacacata ctacattgga
     5101 acatgcagat tgtactttca tggtcgataa tgaggctatt tacgacatgt gcaaaagaaa
     5161 cttggatatc ccaagaccaa gctttgcaaa cttaaacaac ctaattgctc aagtggtatc
     5221 atctgttaca gcatcattga gattcgacgg ttcattaaat gtagatttga acgaatttca
     5281 aaccaatttg gttccatatc caagaattca tttcccctta gtttcatatt ctccagtctt
     5341 atccaaatca aaggcattcc atgagtccaa ctctgtgtca gaaattacaa acgcttgttt
     5401 tgaacctggt aaccagatgg tcaagtgtga tccaagagat ggtaaataca tggctacttg
     5461 tctgttatac aggggtgatg tggtaacaag agatgttcaa agagctgtcg agcaggtgaa
     5521 aaacaagaag accgtccaat tggttgattg gtgtccaact ggtttcaaga tcggtatttg
     5581 ctacgaacct ccaactgcca caccaaactc acaattggcc actgtggata gggccgtctg
     5641 tatgttgtca aataccacat ccattgctga ggcttggaag agaatcgata gaaaattcga
     5701 tttaatgtat gccaaacgtg ctttcgtcca ctggtatgtc ggtgaaggta tggaagaagg
     5761 tgaattcacc gaagctagag aagatttggc tgctttagaa agagattaca tcgaagtggg
     5821 tgccgactca tacgctgagg aagaggaatt ttaaggttca ctctcgcccc caacctcctt
     5881 atcctttcta tatttcaagt atattgtaaa aaaaaaaatt ccccctactg ttatatctct
     5941 ttggtgatat cttatctata atctacttat tatatttctc taatatctat gtacaaaaaa
     6001 taaaaaaaaa aaccattctc cttctctaac ctgcatttga tgaaagctta gtaagtctca
     6061 tcgtagttga tgggccaaag acatttgtac ttgtttcatc agattcttac atcacccatt
     6121 tccatgatta tttcacggat ttcattagta attgcatttt tgtatcccta gttccttttc
     6181 tcccgccttg ccgtcgaatt gaaaaaaaaa atggaaatga ggacatcttc cattgatatc
     6241 caggacaaag caagagtaga agtaactatt agcaaggaga attccaaaaa tagtgaactt
     6301 caaggtaagc agatctatag tttcaatatc tgattttctg cttttcaaaa cttaaaattc
     6361 attagaaggt aaaacaagaa cccgtaaaat taaagttacc gatattatca ggtttttcac
     6421 cccatgtcta ctatcccatt tagaaagaac tatgtgttca aaaactgggc cggtcgacct
     6481 gcagcgtacg aagcttcagc tggcggccgc gttctatagt gtcacctaaa tcgtatgtgt
     6541 atgatacata aggttatgta ttaattgtag ccgcgttcta acgacaatat gtccatatgg
     6601 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcc
//
