LOCUS       sequence-75767-e        6564 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6564)
  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 6564)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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:mEos2-Tub1+3'UTR::TRP1
FEATURES             Location/Qualifiers
     source          1..6564
                     /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             3616..4293
                     /codon_start=1
                     /product="monomeric variant of green-to-red photoswitchable
                     fluorescent protein EosFP (McKinney et al., 2009)"
                     /label=mEos2
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDIHMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     RBS             5778..5786
                     /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(6418..6436)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6419..6436)
                     /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 tcgagatgag tgcgattaag ccagacatga agatcaaact ccgtatggaa
     3661 ggcaacgtaa acgggcacca ctttgtgatc gacggagatg gtacaggcaa gccttttgag
     3721 ggaaaacaga gtatggatct tgaagtcaaa gagggcggac ctctgccttt tgcctttgat
     3781 atcctgacca ctgcattcca ttacggcaac agggtattcg ccaaatatcc agacaacata
     3841 caagactatt ttaagcagtc gtttcctaag gggtattcgt gggaacgaag cttgactttc
     3901 gaagacgggg gcatttgcat tgccagaaac gacataacaa tggaagggga cactttctat
     3961 aataaagttc gattttatgg taccaacttt cccgccaatg gtccagttat gcagaagaag
     4021 acgctgaaat gggagccctc cactgagaaa atgtatgtgc gtgatggagt gctgacgggt
     4081 gatattcata tggctttgtt gcttgaagga aatgcccatt accgatgtga cttcagaact
     4141 acttacaaag ctaaggagaa gggtgtcaag ttaccaggct accactttgt ggaccactgc
     4201 attgagattt taagccatga caaagattac aacaaggtta agctgtatga gcatgctgtt
     4261 gctcattctg gattgcctga caatgccaga cgaggatcca tgagagaagt tattagtatt
     4321 aatggtatgt tcgatttgcc cgtccaggct agatcttttt tttgttagtt cattttggcg
     4381 tttcatcttt cattactaac ttcaacgcaa aatttttttt tttttgattt ctctttacag
     4441 tcggtcaagc tggttgtcag attggtaatg cctgttggga attatattcc cttgagcacg
     4501 gtattaagcc ggatggacat ctagaagatg gcctttcaaa gccgaaggga ggagaagagg
     4561 gtttctccac gtttttccat gaaaccggct acggtaagtt cgttccaagg gctatttacg
     4621 tggatttaga gcccaacgtt attgacgaag tccgtaacgg tccttacaag gacttgttcc
     4681 atccagaaca gttgatcagt ggtaaggagg acgcggctaa taattatgca agaggccatt
     4741 acaccgttgg tagagaaatt ttgggcgatg ttctggatag gattagaaaa ctggcagacc
     4801 aatgtgatgg gttacaaggg ttcttgttta cccattctct tggtggtggt actggttccg
     4861 gtctagggtc cttactgttg gaagaactat ccgctgaata cggtaagaaa tccaagctgg
     4921 aatttgccgt ataccctgct ccacaagtgt ctacttctgt ggttgagcct tacaacacag
     4981 ttttaactac acatactaca ttggaacatg cagattgtac tttcatggtc gataatgagg
     5041 ctatttacga catgtgcaaa agaaacttgg atatcccaag accaagcttt gcaaacttaa
     5101 acaacctaat tgctcaagtg gtatcatctg ttacagcatc attgagattc gacggttcat
     5161 taaatgtaga tttgaacgaa tttcaaacca atttggttcc atatccaaga attcatttcc
     5221 ccttagtttc atattctcca gtcttatcca aatcaaaggc attccatgag tccaactctg
     5281 tgtcagaaat tacaaacgct tgttttgaac ctggtaacca gatggtcaag tgtgatccaa
     5341 gagatggtaa atacatggct acttgtctgt tatacagggg tgatgtggta acaagagatg
     5401 ttcaaagagc tgtcgagcag gtgaaaaaca agaagaccgt ccaattggtt gattggtgtc
     5461 caactggttt caagatcggt atttgctacg aacctccaac tgccacacca aactcacaat
     5521 tggccactgt ggatagggcc gtctgtatgt tgtcaaatac cacatccatt gctgaggctt
     5581 ggaagagaat cgatagaaaa ttcgatttaa tgtatgccaa acgtgctttc gtccactggt
     5641 atgtcggtga aggtatggaa gaaggtgaat tcaccgaagc tagagaagat ttggctgctt
     5701 tagaaagaga ttacatcgaa gtgggtgccg actcatacgc tgaggaagag gaattttaag
     5761 gttcactctc gcccccaacc tccttatcct ttctatattt caagtatatt gtaaaaaaaa
     5821 aaattccccc tactgttata tctctttggt gatatcttat ctataatcta cttattatat
     5881 ttctctaata tctatgtaca aaaaataaaa aaaaaaacca ttctccttct ctaacctgca
     5941 tttgatgaaa gcttagtaag tctcatcgta gttgatgggc caaagacatt tgtacttgtt
     6001 tcatcagatt cttacatcac ccatttccat gattatttca cggatttcat tagtaattgc
     6061 atttttgtat ccctagttcc ttttctcccg ccttgccgtc gaattgaaaa aaaaaatgga
     6121 aatgaggaca tcttccattg atatccagga caaagcaaga gtagaagtaa ctattagcaa
     6181 ggagaattcc aaaaatagtg aacttcaagg taagcagatc tatagtttca atatctgatt
     6241 ttctgctttt caaaacttaa aattcattag aaggtaaaac aagaacccgt aaaattaaag
     6301 ttaccgatat tatcaggttt ttcaccccat gtctactatc ccatttagaa agaactatgt
     6361 gttcaaaaac tgggccggtc gacctgcagc gtacgaagct tcagctggcg gccgcgttct
     6421 atagtgtcac ctaaatcgta tgtgtatgat acataaggtt atgtattaat tgtagccgcg
     6481 ttctaacgac aatatgtcca tatggtgcac tctcagtaca atctgctctg atgccgcata
     6541 gttaagccag ccccgacacc cgcc
//
