LOCUS       sequence-75756-e        7815 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 7815)
  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 7815)
  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::LEU2
FEATURES             Location/Qualifiers
     source          1..7815
                     /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"
     CDS             complement(2776..3870)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(3871..4278)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        4640..4826
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             join(4828..4830,4831..4833,4834..5544)
                     /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(4873..4894)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5134..5153)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5481..5502
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             7029..7037
                     /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(7669..7687)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7670..7687)
                     /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 gcaccatatc gactacgtcg ttaaggccgt ttctgacaga gtaaaattct
     2341 tgagggaact ttcaccatta tgggaaatgg ttcaagaagg tattgactta aactccatca
     2401 aatggtcagg tcattgagtg ttttttattt gttgtatttt ttttttttta gagaaaatcc
     2461 tccaatatat aaattaggaa tcatagtttc atgattttct gttacaccta actttttgtg
     2521 tggtgccctc ctccttgtca atattaatgt taaagtgcaa ttctttttcc ttatcacgtt
     2581 gagccattag tatcaatttg cttacctgta ttcctttaca tcctcctttt tctccttctt
     2641 gataaatgta tgtagattgc gtatatagtt tcgtctaccc tatgaacata ttccattttg
     2701 taatttcgtg tcgtttctat tatgaatttc atttataaag tttatgtaca aatatcataa
     2761 aaaaagagaa tctttttaag caaggatttt cttaacttct tcggcgacag catcaccgac
     2821 ttcggtggta ctgttggaac cacctaaatc accagttctg atacctgcat ccaaaacctt
     2881 tttaactgca tcttcaatgg ccttaccttc ttcaggcaag ttcaatgaca atttcaacat
     2941 cattgcagca gacaagatag tggcgatagg gttgacctta ttctttggca aatctggagc
     3001 agaaccgtgg catggttcgt acaaaccaaa tgcggtgttc ttgtctggca aagaggccaa
     3061 ggacgcagat ggcaacaaac ccaaggaacc tgggataacg gaggcttcat cggagatgat
     3121 atcaccaaac atgttgctgg tgattataat accatttagg tgggttgggt tcttaactag
     3181 gatcatggcg gcagaatcaa tcaattgatg ttgaaccttc aatgtaggga attcgttctt
     3241 gatggtttcc tccacagttt ttctccataa tcttgaagag gccaaaacat tagctttatc
     3301 caaggaccaa ataggcaatg gtggctcatg ttgtagggcc atgaaagcgg ccattcttgt
     3361 gattctttgc acttctggaa cggtgtattg ttcactatcc caagcgacac catcaccatc
     3421 gtcttccttt ctcttaccaa agtaaatacc tcccactaat tctctgacaa caacgaagtc
     3481 agtaccttta gcaaattgtg gcttgattgg agataagtct aaaagagagt cggatgcaaa
     3541 gttacatggt cttaagttgg cgtacaattg aagttcttta cggattttta gtaaaccttg
     3601 ttcaggtcta acactaccgg taccccattt aggaccaccc acagcaccta acaaaacggc
     3661 atcagccttc ttggaggctt ccagcgcctc atctggaagt gggacacctg tagcatcgat
     3721 agcagcacca ccaattaaat gattttcgaa atcgaacttg acattggaac gaacatcaga
     3781 aatagcttta agaaccttaa tggcttcggc tgtgatttct tgaccaacgt ggtcacctgg
     3841 caaaacgacg atcttcttag gggcagacat tagaatggta tatccttgaa atatatatat
     3901 atatattgct gaaatgtaaa aggtaagaaa agttagaaag taagacgatt gctaaccacc
     3961 tattggaaaa aacaataggt ccttaaataa tattgtcaac ttcaagtatt gtgatgcaag
     4021 catttagtca tgaacgcttc tctattctat atgaaaagcc ggttccggcg ctctcacctt
     4081 tcctttttct cccaattttt cagttgaaaa aggtatatgc gtcaggcgac ctctgaaatt
     4141 aacaaaaaat ttccagtcat cgaatttgat tctgtgcgat agcgcccctg tgtgttctcg
     4201 ttatgttgag gaaaaaaata atggttgcta agagattcga actcttgcat cttacgatac
     4261 ctgagtattc ccacagttaa ctgcggtcaa gatatttctt gaatcaggcg ccttagaccg
     4321 ctcggccaaa caaccaatta cttgttgaga aatagagtat aattatccta taaatataac
     4381 gtttttgaac acacatgaac aaggaagtac aggacaattg attttgaaga gaatgtggat
     4441 tttgatgtaa ttgttgggat tccattttta ataaggcaat aatattaggt atgtggatat
     4501 actagaagtt ctcctcgagg gtcgatatgc ggtgtgaaat accgcacaga tgcgtaagga
     4561 gaaaataccg catcaggggc gcgccttaat taaccactaa agggaacaaa agctggtacc
     4621 ctattactct tggcctcctc tagtacactc tatatttttt tatgcctcgg taatgatttt
     4681 catttttttt ttccacctag cggatgactc tttttttttc ttagcgattg gcattatcac
     4741 ataatgaatt atacattata taaagtaatg tgatttcttc gaagaatata ctaaaaaatg
     4801 agcaggcaag ataaacgaag gcaaagcatg gtgagcaagg gcgaggagct gttcaccggg
     4861 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     4921 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     4981 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg cgtgcagtgc
     5041 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     5101 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     5161 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     5221 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actactttag cgacaacgtc
     5281 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     5341 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     5401 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac
     5461 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     5521 ctcggcatgg acgagctgta caagggatcc atgagagaag ttattagtat taatggtatg
     5581 ttcgatttgc ccgtccaggc tagatctttt ttttgttagt tcattttggc gtttcatctt
     5641 tcattactaa cttcaacgca aaattttttt ttttttgatt tctctttaca gtcggtcaag
     5701 ctggttgtca gattggtaat gcctgttggg aattatattc ccttgagcac ggtattaagc
     5761 cggatggaca tctagaagat ggcctttcaa agccgaaggg aggagaagag ggtttctcca
     5821 cgtttttcca tgaaaccggc tacggtaagt tcgttccaag ggctatttac gtggatttag
     5881 agcccaacgt tattgacgaa gtccgtaacg gtccttacaa ggacttgttc catccagaac
     5941 agttgatcag tggtaaggag gacgcggcta ataattatgc aagaggccat tacaccgttg
     6001 gtagagaaat tttgggcgat gttctggata ggattagaaa actggcagac caatgtgatg
     6061 ggttacaagg gttcttgttt acccattctc ttggtggtgg tactggttcc ggtctagggt
     6121 ccttactgtt ggaagaacta tccgctgaat acggtaagaa atccaagctg gaatttgccg
     6181 tataccctgc tccacaagtg tctacttctg tggttgagcc ttacaacaca gttttaacta
     6241 cacatactac attggaacat gcagattgta ctttcatggt cgataatgag gctatttacg
     6301 acatgtgcaa aagaaacttg gatatcccaa gaccaagctt tgcaaactta aacaacctaa
     6361 ttgctcaagt ggtatcatct gttacagcat cattgagatt cgacggttca ttaaatgtag
     6421 atttgaacga atttcaaacc aatttggttc catatccaag aattcatttc cccttagttt
     6481 catattctcc agtcttatcc aaatcaaagg cattccatga gtccaactct gtgtcagaaa
     6541 ttacaaacgc ttgttttgaa cctggtaacc agatggtcaa gtgtgatcca agagatggta
     6601 aatacatggc tacttgtctg ttatacaggg gtgatgtggt aacaagagat gttcaaagag
     6661 ctgtcgagca ggtgaaaaac aagaagaccg tccaattggt tgattggtgt ccaactggtt
     6721 tcaagatcgg tatttgctac gaacctccaa ctgccacacc aaactcacaa ttggccactg
     6781 tggatagggc cgtctgtatg ttgtcaaata ccacatccat tgctgaggct tggaagagaa
     6841 tcgatagaaa attcgattta atgtatgcca aacgtgcttt cgtccactgg tatgtcggtg
     6901 aaggtatgga agaaggtgaa ttcaccgaag ctagagaaga tttggctgct ttagaaagag
     6961 attacatcga agtgggtgcc gactcatacg ctgaggaaga ggaattttaa ggttcactct
     7021 cgcccccaac ctccttatcc tttctatatt tcaagtatat tgtaaaaaaa aaaattcccc
     7081 ctactgttat atctctttgg tgatatctta tctataatct acttattata tttctctaat
     7141 atctatgtac aaaaaataaa aaaaaaaacc attctccttc tctaacctgc atttgatgaa
     7201 agcttagtaa gtctcatcgt agttgatggg ccaaagacat ttgtacttgt ttcatcagat
     7261 tcttacatca cccatttcca tgattatttc acggatttca ttagtaattg catttttgta
     7321 tccctagttc cttttctccc gccttgccgt cgaattgaaa aaaaaaatgg aaatgaggac
     7381 atcttccatt gatatccagg acaaagcaag agtagaagta actattagca aggagaattc
     7441 caaaaatagt gaacttcaag gtaagcagat ctatagtttc aatatctgat tttctgcttt
     7501 tcaaaactta aaattcatta gaaggtaaaa caagaacccg taaaattaaa gttaccgata
     7561 ttatcaggtt tttcacccca tgtctactat cccatttaga aagaactatg tgttcaaaaa
     7621 ctgggccggt cgacctgcag cgtacgaagc ttcagctggc ggccgcgttc tatagtgtca
     7681 cctaaatcgt atgtgtatga tacataaggt tatgtattaa ttgtagccgc gttctaacga
     7741 caatatgtcc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     7801 gccccgacac ccgcc
//
