LOCUS       sequence-75752-e        6689 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 6689)
  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 6689)
  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::URA3
FEATURES             Location/Qualifiers
     source          1..6689
                     /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
     RBS             5903..5911
                     /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(6543..6561)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6544..6561)
                     /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 atggtaagta
     3721 aaggtgaaga gacaactatg ggggtaatca aaccagacat gaagatcaaa ctgaaaatgg
     3781 aaggtaacgt taatggccac gcgtttgtta ttgaaggtga aggagagggg aaaccttacg
     3841 acggaaccaa cactatcaat ttggaggtta aggaaggtgc accattacca ttcagctatg
     3901 acatcttaac aactgcattt tcatacggca acagagcttt taccaagtac cctgacgata
     3961 taccaaatta cttcaaacaa tcctttcctg aaggttattc atgggagaga acaatgacgt
     4021 ttgaagataa gggcattgtc aaggtcaaat ctgacatttc tatggaagag gattcattca
     4081 tctacgagat tcacctcaag ggcgaaaact tcccaccaaa tggtccagtg atgcaaaagg
     4141 aaactactgg atgggatgct tctacagaaa ggatgtacgt gagagatgga gtcttaaaag
     4201 gtgatgttaa aatgaagttg ctacttgaag gtggtggcca tcatagagtt gatttcaaaa
     4261 caatctacag agccaaaaag gctgtaaaac ttcctgatta tcattttgtg gatcatcgta
     4321 tagaaattct aaaccacgac aaagattaca ataaggtcac tgtttatgaa atagcagttg
     4381 ccagaaattc tacagatgga atggatgaat tgtacaaagg atccatgaga gaagttatta
     4441 gtattaatgg tatgttcgat ttgcccgtcc aggctagatc ttttttttgt tagttcattt
     4501 tggcgtttca tctttcatta ctaacttcaa cgcaaaattt tttttttttt gatttctctt
     4561 tacagtcggt caagctggtt gtcagattgg taatgcctgt tgggaattat attcccttga
     4621 gcacggtatt aagccggatg gacatctaga agatggcctt tcaaagccga agggaggaga
     4681 agagggtttc tccacgtttt tccatgaaac cggctacggt aagttcgttc caagggctat
     4741 ttacgtggat ttagagccca acgttattga cgaagtccgt aacggtcctt acaaggactt
     4801 gttccatcca gaacagttga tcagtggtaa ggaggacgcg gctaataatt atgcaagagg
     4861 ccattacacc gttggtagag aaattttggg cgatgttctg gataggatta gaaaactggc
     4921 agaccaatgt gatgggttac aagggttctt gtttacccat tctcttggtg gtggtactgg
     4981 ttccggtcta gggtccttac tgttggaaga actatccgct gaatacggta agaaatccaa
     5041 gctggaattt gccgtatacc ctgctccaca agtgtctact tctgtggttg agccttacaa
     5101 cacagtttta actacacata ctacattgga acatgcagat tgtactttca tggtcgataa
     5161 tgaggctatt tacgacatgt gcaaaagaaa cttggatatc ccaagaccaa gctttgcaaa
     5221 cttaaacaac ctaattgctc aagtggtatc atctgttaca gcatcattga gattcgacgg
     5281 ttcattaaat gtagatttga acgaatttca aaccaatttg gttccatatc caagaattca
     5341 tttcccctta gtttcatatt ctccagtctt atccaaatca aaggcattcc atgagtccaa
     5401 ctctgtgtca gaaattacaa acgcttgttt tgaacctggt aaccagatgg tcaagtgtga
     5461 tccaagagat ggtaaataca tggctacttg tctgttatac aggggtgatg tggtaacaag
     5521 agatgttcaa agagctgtcg agcaggtgaa aaacaagaag accgtccaat tggttgattg
     5581 gtgtccaact ggtttcaaga tcggtatttg ctacgaacct ccaactgcca caccaaactc
     5641 acaattggcc actgtggata gggccgtctg tatgttgtca aataccacat ccattgctga
     5701 ggcttggaag agaatcgata gaaaattcga tttaatgtat gccaaacgtg ctttcgtcca
     5761 ctggtatgtc ggtgaaggta tggaagaagg tgaattcacc gaagctagag aagatttggc
     5821 tgctttagaa agagattaca tcgaagtggg tgccgactca tacgctgagg aagaggaatt
     5881 ttaaggttca ctctcgcccc caacctcctt atcctttcta tatttcaagt atattgtaaa
     5941 aaaaaaaatt ccccctactg ttatatctct ttggtgatat cttatctata atctacttat
     6001 tatatttctc taatatctat gtacaaaaaa taaaaaaaaa aaccattctc cttctctaac
     6061 ctgcatttga tgaaagctta gtaagtctca tcgtagttga tgggccaaag acatttgtac
     6121 ttgtttcatc agattcttac atcacccatt tccatgatta tttcacggat ttcattagta
     6181 attgcatttt tgtatcccta gttccttttc tcccgccttg ccgtcgaatt gaaaaaaaaa
     6241 atggaaatga ggacatcttc cattgatatc caggacaaag caagagtaga agtaactatt
     6301 agcaaggaga attccaaaaa tagtgaactt caaggtaagc agatctatag tttcaatatc
     6361 tgattttctg cttttcaaaa cttaaaattc attagaaggt aaaacaagaa cccgtaaaat
     6421 taaagttacc gatattatca ggtttttcac cccatgtcta ctatcccatt tagaaagaac
     6481 tatgtgttca aaaactgggc cggtcgacct gcagcgtacg aagcttcagc tggcggccgc
     6541 gttctatagt gtcacctaaa tcgtatgtgt atgatacata aggttatgta ttaattgtag
     6601 ccgcgttcta acgacaatat gtccatatgg tgcactctca gtacaatctg ctctgatgcc
     6661 gcatagttaa gccagccccg acacccgcc
//
