LOCUS       sequence-75765-e        6595 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 6595)
  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 6595)
  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:Venus-Tub1+3'UTR::TRP1
FEATURES             Location/Qualifiers
     source          1..6595
                     /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        2303..2583
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             2584..3258
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        3423..3609
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3611..4324
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     RBS             5809..5817
                     /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(6449..6467)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6450..6467)
                     /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 gcaccataga tcacgacatt actatatata taatatagga agcatttaat
     2341 agaacagcat cgtaatatat gtgtactttg cagttatgac gccagatggc agtagtggaa
     2401 gatattcttt attgaaaaat agcttgtcac cttacgtaca atcttgatcc ggagcttttc
     2461 tttttttgcc gattaagaat taattcggtc gaaaaaagaa aaggagaggg ccaagaggga
     2521 gggcattggt gactattgag cacgtgagta tacgtgatta agcacacaaa ggcagcttgg
     2581 agtatgtctg ttattaattt cacaggtagt tctggtccat tggtgaaagt ttgcggcttg
     2641 cagagcacag aggccgcaga atgtgctcta gattccgatg ctgacttgct gggtattata
     2701 tgtgtgccca atagaaagag aacaattgac ccggttattg caaggaaaat ttcaagtctt
     2761 gtaaaagcat ataaaaatag ttcaggcact ccgaaatact tggttggcgt gtttcgtaat
     2821 caacctaagg aggatgtttt ggctctggtc aatgattacg gcattgatat cgtccaactg
     2881 catggagatg agtcgtggca agaataccaa gagttcctcg gtttgccagt tattaaaaga
     2941 ctcgtatttc caaaagactg caacatacta ctcagtgcag cttcacagaa acctcattcg
     3001 tttattccct tgtttgattc agaagcaggt gggacaggtg aacttttgga ttggaactcg
     3061 atttctgact gggttggaag gcaagagagc cccgaaagct tacattttat gttagctggt
     3121 ggactgacgc cagaaaatgt tggtgatgcg cttagattaa atggcgttat tggtgttgat
     3181 gtaagcggag gtgtggagac aaatggtgta aaagactcta acaaaatagc aaatttcgtc
     3241 aaaaatgcta agaaataggt tattactgag tagtatttat ttaagtattg tttgtgcact
     3301 tgccgatcta tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     3361 ggcgcgcctt aattaaccac taaagggaac aaaagctggt accctattac tcttggcctc
     3421 ctctagtaca ctctatattt ttttatgcct cggtaatgat tttcattttt tttttccacc
     3481 tagcggatga ctcttttttt ttcttagcga ttggcattat cacataatga attatacatt
     3541 atataaagta atgtgatttc ttcgaagaat atactaaaaa atgagcaggc aagataaacg
     3601 aaggcaaagc atgtctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga
     3661 attagatggt gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc
     3721 tacttacggt aaattgacct taaaattgat ttgtactact ggtaaattgc cagttccatg
     3781 gccaacctta gtcactactt taggttatgg tttgcaatgt tttgctagat acccagatca
     3841 tatgaaacaa catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac
     3901 tatttttttc aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga
     3961 taccttagtt aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt
     4021 aggtcacaaa ttggaataca actataactc tcacaatgtt tacatcactg ctgacaaaca
     4081 aaagaatggt atcaaagcta acttcaaaat tagacacaac attgaagatg gtggtgttca
     4141 attagctgac cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga
     4201 caaccattac ttatcctatc aatctgcctt atccaaagat ccaaacgaaa agagagacca
     4261 catggtcttg ttagaatttg ttactgctgc tggtattacc catggtatgg atgaattgta
     4321 caaaggatcc atgagagaag ttattagtat taatggtatg ttcgatttgc ccgtccaggc
     4381 tagatctttt ttttgttagt tcattttggc gtttcatctt tcattactaa cttcaacgca
     4441 aaattttttt ttttttgatt tctctttaca gtcggtcaag ctggttgtca gattggtaat
     4501 gcctgttggg aattatattc ccttgagcac ggtattaagc cggatggaca tctagaagat
     4561 ggcctttcaa agccgaaggg aggagaagag ggtttctcca cgtttttcca tgaaaccggc
     4621 tacggtaagt tcgttccaag ggctatttac gtggatttag agcccaacgt tattgacgaa
     4681 gtccgtaacg gtccttacaa ggacttgttc catccagaac agttgatcag tggtaaggag
     4741 gacgcggcta ataattatgc aagaggccat tacaccgttg gtagagaaat tttgggcgat
     4801 gttctggata ggattagaaa actggcagac caatgtgatg ggttacaagg gttcttgttt
     4861 acccattctc ttggtggtgg tactggttcc ggtctagggt ccttactgtt ggaagaacta
     4921 tccgctgaat acggtaagaa atccaagctg gaatttgccg tataccctgc tccacaagtg
     4981 tctacttctg tggttgagcc ttacaacaca gttttaacta cacatactac attggaacat
     5041 gcagattgta ctttcatggt cgataatgag gctatttacg acatgtgcaa aagaaacttg
     5101 gatatcccaa gaccaagctt tgcaaactta aacaacctaa ttgctcaagt ggtatcatct
     5161 gttacagcat cattgagatt cgacggttca ttaaatgtag atttgaacga atttcaaacc
     5221 aatttggttc catatccaag aattcatttc cccttagttt catattctcc agtcttatcc
     5281 aaatcaaagg cattccatga gtccaactct gtgtcagaaa ttacaaacgc ttgttttgaa
     5341 cctggtaacc agatggtcaa gtgtgatcca agagatggta aatacatggc tacttgtctg
     5401 ttatacaggg gtgatgtggt aacaagagat gttcaaagag ctgtcgagca ggtgaaaaac
     5461 aagaagaccg tccaattggt tgattggtgt ccaactggtt tcaagatcgg tatttgctac
     5521 gaacctccaa ctgccacacc aaactcacaa ttggccactg tggatagggc cgtctgtatg
     5581 ttgtcaaata ccacatccat tgctgaggct tggaagagaa tcgatagaaa attcgattta
     5641 atgtatgcca aacgtgcttt cgtccactgg tatgtcggtg aaggtatgga agaaggtgaa
     5701 ttcaccgaag ctagagaaga tttggctgct ttagaaagag attacatcga agtgggtgcc
     5761 gactcatacg ctgaggaaga ggaattttaa ggttcactct cgcccccaac ctccttatcc
     5821 tttctatatt tcaagtatat tgtaaaaaaa aaaattcccc ctactgttat atctctttgg
     5881 tgatatctta tctataatct acttattata tttctctaat atctatgtac aaaaaataaa
     5941 aaaaaaaacc attctccttc tctaacctgc atttgatgaa agcttagtaa gtctcatcgt
     6001 agttgatggg ccaaagacat ttgtacttgt ttcatcagat tcttacatca cccatttcca
     6061 tgattatttc acggatttca ttagtaattg catttttgta tccctagttc cttttctccc
     6121 gccttgccgt cgaattgaaa aaaaaaatgg aaatgaggac atcttccatt gatatccagg
     6181 acaaagcaag agtagaagta actattagca aggagaattc caaaaatagt gaacttcaag
     6241 gtaagcagat ctatagtttc aatatctgat tttctgcttt tcaaaactta aaattcatta
     6301 gaaggtaaaa caagaacccg taaaattaaa gttaccgata ttatcaggtt tttcacccca
     6361 tgtctactat cccatttaga aagaactatg tgttcaaaaa ctgggccggt cgacctgcag
     6421 cgtacgaagc ttcagctggc ggccgcgttc tatagtgtca cctaaatcgt atgtgtatga
     6481 tacataaggt tatgtattaa ttgtagccgc gttctaacga caatatgtcc atatggtgca
     6541 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgcc
//
