LOCUS       sequence-75750-e        6693 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 6693)
  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 6693)
  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:mTurquoise2-Tub1+3'UTR::URA3
FEATURES             Location/Qualifiers
     source          1..6693
                     /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(3706..3708,3709..3711,3712..4422)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3751..3772)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4012..4031)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4359..4380
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             5907..5915
                     /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(6547..6565)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6548..6565)
                     /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 aaagcatggt gagcaagggc
     3721 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     3781 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     3841 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     3901 tcctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     3961 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     4021 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     4081 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     4141 tactttagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     4201 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     4261 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     4321 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     4381 accgccgccg ggatcactct cggcatggac gagctgtaca agggatccat gagagaagtt
     4441 attagtatta atggtatgtt cgatttgccc gtccaggcta gatctttttt ttgttagttc
     4501 attttggcgt ttcatctttc attactaact tcaacgcaaa attttttttt ttttgatttc
     4561 tctttacagt cggtcaagct ggttgtcaga ttggtaatgc ctgttgggaa ttatattccc
     4621 ttgagcacgg tattaagccg gatggacatc tagaagatgg cctttcaaag ccgaagggag
     4681 gagaagaggg tttctccacg tttttccatg aaaccggcta cggtaagttc gttccaaggg
     4741 ctatttacgt ggatttagag cccaacgtta ttgacgaagt ccgtaacggt ccttacaagg
     4801 acttgttcca tccagaacag ttgatcagtg gtaaggagga cgcggctaat aattatgcaa
     4861 gaggccatta caccgttggt agagaaattt tgggcgatgt tctggatagg attagaaaac
     4921 tggcagacca atgtgatggg ttacaagggt tcttgtttac ccattctctt ggtggtggta
     4981 ctggttccgg tctagggtcc ttactgttgg aagaactatc cgctgaatac ggtaagaaat
     5041 ccaagctgga atttgccgta taccctgctc cacaagtgtc tacttctgtg gttgagcctt
     5101 acaacacagt tttaactaca catactacat tggaacatgc agattgtact ttcatggtcg
     5161 ataatgaggc tatttacgac atgtgcaaaa gaaacttgga tatcccaaga ccaagctttg
     5221 caaacttaaa caacctaatt gctcaagtgg tatcatctgt tacagcatca ttgagattcg
     5281 acggttcatt aaatgtagat ttgaacgaat ttcaaaccaa tttggttcca tatccaagaa
     5341 ttcatttccc cttagtttca tattctccag tcttatccaa atcaaaggca ttccatgagt
     5401 ccaactctgt gtcagaaatt acaaacgctt gttttgaacc tggtaaccag atggtcaagt
     5461 gtgatccaag agatggtaaa tacatggcta cttgtctgtt atacaggggt gatgtggtaa
     5521 caagagatgt tcaaagagct gtcgagcagg tgaaaaacaa gaagaccgtc caattggttg
     5581 attggtgtcc aactggtttc aagatcggta tttgctacga acctccaact gccacaccaa
     5641 actcacaatt ggccactgtg gatagggccg tctgtatgtt gtcaaatacc acatccattg
     5701 ctgaggcttg gaagagaatc gatagaaaat tcgatttaat gtatgccaaa cgtgctttcg
     5761 tccactggta tgtcggtgaa ggtatggaag aaggtgaatt caccgaagct agagaagatt
     5821 tggctgcttt agaaagagat tacatcgaag tgggtgccga ctcatacgct gaggaagagg
     5881 aattttaagg ttcactctcg cccccaacct ccttatcctt tctatatttc aagtatattg
     5941 taaaaaaaaa aattccccct actgttatat ctctttggtg atatcttatc tataatctac
     6001 ttattatatt tctctaatat ctatgtacaa aaaataaaaa aaaaaaccat tctccttctc
     6061 taacctgcat ttgatgaaag cttagtaagt ctcatcgtag ttgatgggcc aaagacattt
     6121 gtacttgttt catcagattc ttacatcacc catttccatg attatttcac ggatttcatt
     6181 agtaattgca tttttgtatc cctagttcct tttctcccgc cttgccgtcg aattgaaaaa
     6241 aaaaatggaa atgaggacat cttccattga tatccaggac aaagcaagag tagaagtaac
     6301 tattagcaag gagaattcca aaaatagtga acttcaaggt aagcagatct atagtttcaa
     6361 tatctgattt tctgcttttc aaaacttaaa attcattaga aggtaaaaca agaacccgta
     6421 aaattaaagt taccgatatt atcaggtttt tcaccccatg tctactatcc catttagaaa
     6481 gaactatgtg ttcaaaaact gggccggtcg acctgcagcg tacgaagctt cagctggcgg
     6541 ccgcgttcta tagtgtcacc taaatcgtat gtgtatgata cataaggtta tgtattaatt
     6601 gtagccgcgt tctaacgaca atatgtccat atggtgcact ctcagtacaa tctgctctga
     6661 tgccgcatag ttaagccagc cccgacaccc gcc
//
