LOCUS       sequence-75751-e        6698 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 6698)
  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 6698)
  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:CloverGFP-Tub1+3'UTR::URA3
FEATURES             Location/Qualifiers
     source          1..6698
                     /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             join(3711..3713,3714..3716,3717..4427)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3756..3777)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     4364..4385
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             5912..5920
                     /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(6552..6570)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6553..6570)
                     /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 atggtgagca
     3721 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     3781 acggccacaa gttcagcgtc cgcggcgagg gcgagggcga tgccaccaac ggcaagctga
     3841 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     3901 ccttcggcta cggcgtggcc tgcttcagcc gctaccccga ccacatgaag cagcacgact
     3961 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctct ttcaaggacg
     4021 acggtaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     4081 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     4141 acaacttcaa cagccacaac gtctatatca cggccgacaa gcagaagaac ggcatcaagg
     4201 ctaacttcaa gatccgccac aacgttgagg acggcagcgt gcagctcgcc gaccactacc
     4261 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcc
     4321 atcagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     4381 tcgtgaccgc cgccgggatt acacatggca tggacgagct gtacaaggga tccatgagag
     4441 aagttattag tattaatggt atgttcgatt tgcccgtcca ggctagatct tttttttgtt
     4501 agttcatttt ggcgtttcat ctttcattac taacttcaac gcaaaatttt tttttttttg
     4561 atttctcttt acagtcggtc aagctggttg tcagattggt aatgcctgtt gggaattata
     4621 ttcccttgag cacggtatta agccggatgg acatctagaa gatggccttt caaagccgaa
     4681 gggaggagaa gagggtttct ccacgttttt ccatgaaacc ggctacggta agttcgttcc
     4741 aagggctatt tacgtggatt tagagcccaa cgttattgac gaagtccgta acggtcctta
     4801 caaggacttg ttccatccag aacagttgat cagtggtaag gaggacgcgg ctaataatta
     4861 tgcaagaggc cattacaccg ttggtagaga aattttgggc gatgttctgg ataggattag
     4921 aaaactggca gaccaatgtg atgggttaca agggttcttg tttacccatt ctcttggtgg
     4981 tggtactggt tccggtctag ggtccttact gttggaagaa ctatccgctg aatacggtaa
     5041 gaaatccaag ctggaatttg ccgtataccc tgctccacaa gtgtctactt ctgtggttga
     5101 gccttacaac acagttttaa ctacacatac tacattggaa catgcagatt gtactttcat
     5161 ggtcgataat gaggctattt acgacatgtg caaaagaaac ttggatatcc caagaccaag
     5221 ctttgcaaac ttaaacaacc taattgctca agtggtatca tctgttacag catcattgag
     5281 attcgacggt tcattaaatg tagatttgaa cgaatttcaa accaatttgg ttccatatcc
     5341 aagaattcat ttccccttag tttcatattc tccagtctta tccaaatcaa aggcattcca
     5401 tgagtccaac tctgtgtcag aaattacaaa cgcttgtttt gaacctggta accagatggt
     5461 caagtgtgat ccaagagatg gtaaatacat ggctacttgt ctgttataca ggggtgatgt
     5521 ggtaacaaga gatgttcaaa gagctgtcga gcaggtgaaa aacaagaaga ccgtccaatt
     5581 ggttgattgg tgtccaactg gtttcaagat cggtatttgc tacgaacctc caactgccac
     5641 accaaactca caattggcca ctgtggatag ggccgtctgt atgttgtcaa ataccacatc
     5701 cattgctgag gcttggaaga gaatcgatag aaaattcgat ttaatgtatg ccaaacgtgc
     5761 tttcgtccac tggtatgtcg gtgaaggtat ggaagaaggt gaattcaccg aagctagaga
     5821 agatttggct gctttagaaa gagattacat cgaagtgggt gccgactcat acgctgagga
     5881 agaggaattt taaggttcac tctcgccccc aacctcctta tcctttctat atttcaagta
     5941 tattgtaaaa aaaaaaattc cccctactgt tatatctctt tggtgatatc ttatctataa
     6001 tctacttatt atatttctct aatatctatg tacaaaaaat aaaaaaaaaa accattctcc
     6061 ttctctaacc tgcatttgat gaaagcttag taagtctcat cgtagttgat gggccaaaga
     6121 catttgtact tgtttcatca gattcttaca tcacccattt ccatgattat ttcacggatt
     6181 tcattagtaa ttgcattttt gtatccctag ttccttttct cccgccttgc cgtcgaattg
     6241 aaaaaaaaaa tggaaatgag gacatcttcc attgatatcc aggacaaagc aagagtagaa
     6301 gtaactatta gcaaggagaa ttccaaaaat agtgaacttc aaggtaagca gatctatagt
     6361 ttcaatatct gattttctgc ttttcaaaac ttaaaattca ttagaaggta aaacaagaac
     6421 ccgtaaaatt aaagttaccg atattatcag gtttttcacc ccatgtctac tatcccattt
     6481 agaaagaact atgtgttcaa aaactgggcc ggtcgacctg cagcgtacga agcttcagct
     6541 ggcggccgcg ttctatagtg tcacctaaat cgtatgtgta tgatacataa ggttatgtat
     6601 taattgtagc cgcgttctaa cgacaatatg tccatatggt gcactctcag tacaatctgc
     6661 tctgatgccg catagttaag ccagccccga cacccgcc
//
