LOCUS       sequence-75764-e        6594 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 6594)
  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 6594)
  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::TRP1
FEATURES             Location/Qualifiers
     source          1..6594
                     /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
     RBS             5808..5816
                     /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(6448..6466)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6449..6466)
                     /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 tcgagatggt aagtaaaggt gaagagacaa ctatgggggt aatcaaacca
     3661 gacatgaaga tcaaactgaa aatggaaggt aacgttaatg gccacgcgtt tgttattgaa
     3721 ggtgaaggag aggggaaacc ttacgacgga accaacacta tcaatttgga ggttaaggaa
     3781 ggtgcaccat taccattcag ctatgacatc ttaacaactg cattttcata cggcaacaga
     3841 gcttttacca agtaccctga cgatatacca aattacttca aacaatcctt tcctgaaggt
     3901 tattcatggg agagaacaat gacgtttgaa gataagggca ttgtcaaggt caaatctgac
     3961 atttctatgg aagaggattc attcatctac gagattcacc tcaagggcga aaacttccca
     4021 ccaaatggtc cagtgatgca aaaggaaact actggatggg atgcttctac agaaaggatg
     4081 tacgtgagag atggagtctt aaaaggtgat gttaaaatga agttgctact tgaaggtggt
     4141 ggccatcata gagttgattt caaaacaatc tacagagcca aaaaggctgt aaaacttcct
     4201 gattatcatt ttgtggatca tcgtatagaa attctaaacc acgacaaaga ttacaataag
     4261 gtcactgttt atgaaatagc agttgccaga aattctacag atggaatgga tgaattgtac
     4321 aaaggatcca tgagagaagt tattagtatt aatggtatgt tcgatttgcc cgtccaggct
     4381 agatcttttt tttgttagtt cattttggcg tttcatcttt cattactaac ttcaacgcaa
     4441 aatttttttt tttttgattt ctctttacag tcggtcaagc tggttgtcag attggtaatg
     4501 cctgttggga attatattcc cttgagcacg gtattaagcc ggatggacat ctagaagatg
     4561 gcctttcaaa gccgaaggga ggagaagagg gtttctccac gtttttccat gaaaccggct
     4621 acggtaagtt cgttccaagg gctatttacg tggatttaga gcccaacgtt attgacgaag
     4681 tccgtaacgg tccttacaag gacttgttcc atccagaaca gttgatcagt ggtaaggagg
     4741 acgcggctaa taattatgca agaggccatt acaccgttgg tagagaaatt ttgggcgatg
     4801 ttctggatag gattagaaaa ctggcagacc aatgtgatgg gttacaaggg ttcttgttta
     4861 cccattctct tggtggtggt actggttccg gtctagggtc cttactgttg gaagaactat
     4921 ccgctgaata cggtaagaaa tccaagctgg aatttgccgt ataccctgct ccacaagtgt
     4981 ctacttctgt ggttgagcct tacaacacag ttttaactac acatactaca ttggaacatg
     5041 cagattgtac tttcatggtc gataatgagg ctatttacga catgtgcaaa agaaacttgg
     5101 atatcccaag accaagcttt gcaaacttaa acaacctaat tgctcaagtg gtatcatctg
     5161 ttacagcatc attgagattc gacggttcat taaatgtaga tttgaacgaa tttcaaacca
     5221 atttggttcc atatccaaga attcatttcc ccttagtttc atattctcca gtcttatcca
     5281 aatcaaaggc attccatgag tccaactctg tgtcagaaat tacaaacgct tgttttgaac
     5341 ctggtaacca gatggtcaag tgtgatccaa gagatggtaa atacatggct acttgtctgt
     5401 tatacagggg tgatgtggta acaagagatg ttcaaagagc tgtcgagcag gtgaaaaaca
     5461 agaagaccgt ccaattggtt gattggtgtc caactggttt caagatcggt atttgctacg
     5521 aacctccaac tgccacacca aactcacaat tggccactgt ggatagggcc gtctgtatgt
     5581 tgtcaaatac cacatccatt gctgaggctt ggaagagaat cgatagaaaa ttcgatttaa
     5641 tgtatgccaa acgtgctttc gtccactggt atgtcggtga aggtatggaa gaaggtgaat
     5701 tcaccgaagc tagagaagat ttggctgctt tagaaagaga ttacatcgaa gtgggtgccg
     5761 actcatacgc tgaggaagag gaattttaag gttcactctc gcccccaacc tccttatcct
     5821 ttctatattt caagtatatt gtaaaaaaaa aaattccccc tactgttata tctctttggt
     5881 gatatcttat ctataatcta cttattatat ttctctaata tctatgtaca aaaaataaaa
     5941 aaaaaaacca ttctccttct ctaacctgca tttgatgaaa gcttagtaag tctcatcgta
     6001 gttgatgggc caaagacatt tgtacttgtt tcatcagatt cttacatcac ccatttccat
     6061 gattatttca cggatttcat tagtaattgc atttttgtat ccctagttcc ttttctcccg
     6121 ccttgccgtc gaattgaaaa aaaaaatgga aatgaggaca tcttccattg atatccagga
     6181 caaagcaaga gtagaagtaa ctattagcaa ggagaattcc aaaaatagtg aacttcaagg
     6241 taagcagatc tatagtttca atatctgatt ttctgctttt caaaacttaa aattcattag
     6301 aaggtaaaac aagaacccgt aaaattaaag ttaccgatat tatcaggttt ttcaccccat
     6361 gtctactatc ccatttagaa agaactatgt gttcaaaaac tgggccggtc gacctgcagc
     6421 gtacgaagct tcagctggcg gccgcgttct atagtgtcac ctaaatcgta tgtgtatgat
     6481 acataaggtt atgtattaat tgtagccgcg ttctaacgac aatatgtcca tatggtgcac
     6541 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgcc
//
