LOCUS       sequence-75762-e        6598 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 6598)
  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 6598)
  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::TRP1
FEATURES             Location/Qualifiers
     source          1..6598
                     /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             join(3611..3613,3614..3616,3617..4327)
                     /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(3656..3677)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3917..3936)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4264..4285
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             5812..5820
                     /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(6452..6470)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6453..6470)
                     /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 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     3661 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     3721 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     3781 ctggcccacc ctcgtgacca ccctgtcctg gggcgtgcag tgcttcgccc gctaccccga
     3841 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     3901 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     3961 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     4021 cctggggcac aagctggagt acaactactt tagcgacaac gtctatatca ccgccgacaa
     4081 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt
     4141 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     4201 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
     4261 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     4321 gtacaaggga tccatgagag aagttattag tattaatggt atgttcgatt tgcccgtcca
     4381 ggctagatct tttttttgtt agttcatttt ggcgtttcat ctttcattac taacttcaac
     4441 gcaaaatttt tttttttttg atttctcttt acagtcggtc aagctggttg tcagattggt
     4501 aatgcctgtt gggaattata ttcccttgag cacggtatta agccggatgg acatctagaa
     4561 gatggccttt caaagccgaa gggaggagaa gagggtttct ccacgttttt ccatgaaacc
     4621 ggctacggta agttcgttcc aagggctatt tacgtggatt tagagcccaa cgttattgac
     4681 gaagtccgta acggtcctta caaggacttg ttccatccag aacagttgat cagtggtaag
     4741 gaggacgcgg ctaataatta tgcaagaggc cattacaccg ttggtagaga aattttgggc
     4801 gatgttctgg ataggattag aaaactggca gaccaatgtg atgggttaca agggttcttg
     4861 tttacccatt ctcttggtgg tggtactggt tccggtctag ggtccttact gttggaagaa
     4921 ctatccgctg aatacggtaa gaaatccaag ctggaatttg ccgtataccc tgctccacaa
     4981 gtgtctactt ctgtggttga gccttacaac acagttttaa ctacacatac tacattggaa
     5041 catgcagatt gtactttcat ggtcgataat gaggctattt acgacatgtg caaaagaaac
     5101 ttggatatcc caagaccaag ctttgcaaac ttaaacaacc taattgctca agtggtatca
     5161 tctgttacag catcattgag attcgacggt tcattaaatg tagatttgaa cgaatttcaa
     5221 accaatttgg ttccatatcc aagaattcat ttccccttag tttcatattc tccagtctta
     5281 tccaaatcaa aggcattcca tgagtccaac tctgtgtcag aaattacaaa cgcttgtttt
     5341 gaacctggta accagatggt caagtgtgat ccaagagatg gtaaatacat ggctacttgt
     5401 ctgttataca ggggtgatgt ggtaacaaga gatgttcaaa gagctgtcga gcaggtgaaa
     5461 aacaagaaga ccgtccaatt ggttgattgg tgtccaactg gtttcaagat cggtatttgc
     5521 tacgaacctc caactgccac accaaactca caattggcca ctgtggatag ggccgtctgt
     5581 atgttgtcaa ataccacatc cattgctgag gcttggaaga gaatcgatag aaaattcgat
     5641 ttaatgtatg ccaaacgtgc tttcgtccac tggtatgtcg gtgaaggtat ggaagaaggt
     5701 gaattcaccg aagctagaga agatttggct gctttagaaa gagattacat cgaagtgggt
     5761 gccgactcat acgctgagga agaggaattt taaggttcac tctcgccccc aacctcctta
     5821 tcctttctat atttcaagta tattgtaaaa aaaaaaattc cccctactgt tatatctctt
     5881 tggtgatatc ttatctataa tctacttatt atatttctct aatatctatg tacaaaaaat
     5941 aaaaaaaaaa accattctcc ttctctaacc tgcatttgat gaaagcttag taagtctcat
     6001 cgtagttgat gggccaaaga catttgtact tgtttcatca gattcttaca tcacccattt
     6061 ccatgattat ttcacggatt tcattagtaa ttgcattttt gtatccctag ttccttttct
     6121 cccgccttgc cgtcgaattg aaaaaaaaaa tggaaatgag gacatcttcc attgatatcc
     6181 aggacaaagc aagagtagaa gtaactatta gcaaggagaa ttccaaaaat agtgaacttc
     6241 aaggtaagca gatctatagt ttcaatatct gattttctgc ttttcaaaac ttaaaattca
     6301 ttagaaggta aaacaagaac ccgtaaaatt aaagttaccg atattatcag gtttttcacc
     6361 ccatgtctac tatcccattt agaaagaact atgtgttcaa aaactgggcc ggtcgacctg
     6421 cagcgtacga agcttcagct ggcggccgcg ttctatagtg tcacctaaat cgtatgtgta
     6481 tgatacataa ggttatgtat taattgtagc cgcgttctaa cgacaatatg tccatatggt
     6541 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgcc
//
