LOCUS       sequence-75768-e        6776 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 6776)
  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 6776)
  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::HIS3
FEATURES             Location/Qualifiers
     source          1..6776
                     /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        2429..2616
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             2617..3279
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        3601..3787
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             join(3789..3791,3792..3794,3795..4505)
                     /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(3834..3855)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4095..4114)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4442..4463
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             5990..5998
                     /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(6630..6648)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6631..6648)
                     /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 gcaccataaa ttcccgtttt aagagcttgg tgagcgctag gagtcactgc
     2341 caggtatcgt ttgaacacgg cattagtcag ggaagtcata acacagtcct ttcccgcaat
     2401 tttctttttc tattactctt ggcctcctct agtacactct atattttttt atgcctcggt
     2461 aatgattttc attttttttt ttccacctag cggatgactc tttttttttc ttagcgattg
     2521 gcattatcac ataatgaatt atacattata taaagtaatg tgatttcttc gaagaatata
     2581 ctaaaaaatg agcaggcaag ataaacgaag gcaaagatga cagagcagaa agccctagta
     2641 aagcgtatta caaatgaaac caagattcag attgcgatct ctttaaaggg tggtccccta
     2701 gcgatagagc actcgatctt cccagaaaaa gaggcagaag cagtagcaga acaggccaca
     2761 caatcgcaag tgattaacgt ccacacaggt atagggtttc tggaccatat gatacatgct
     2821 ctggccaagc attccggctg gtcgctaatc gttgagtgca ttggtgactt acacatagac
     2881 gaccatcaca ccactgaaga ctgcgggatt gctctcggtc aagcttttaa agaggcccta
     2941 ggggccgtgc gtggagtaaa aaggtttgga tcaggatttg cgcctttgga tgaggcactt
     3001 tccagagcgg tggtagatct ttcgaacagg ccgtacgcag ttgtcgaact tggtttgcaa
     3061 agggagaaag taggagatct ctcttgcgag atgatcccgc attttcttga aagctttgca
     3121 gaggctagca gaattaccct ccacgttgat tgtctgcgag gcaagaatga tcatcaccgt
     3181 agtgagagtg cgttcaaggc tcttgcggtt gccataagag aagccacctc gcccaatggt
     3241 accaacgatg ttccctccac caaaggtgtt cttatgtagt gacaccgatt atttaaagct
     3301 gcagcatacg atatatatac atgtgtatat atgtatacct atgaatgtca gtaagtatgt
     3361 atacgaacag tatgatactg aagatgacaa ggtaatgcat cattctatac gtgtcattct
     3421 gaacgaggcg cgctttcctt ttttcttttt gctttttctt tttttttctc ttgaactcga
     3481 cggatctatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcagggg
     3541 cgcgccttaa ttaaccacta aagggaacaa aagctggtac cctattactc ttggcctcct
     3601 ctagtacact ctatattttt ttatgcctcg gtaatgattt tcattttttt tttccaccta
     3661 gcggatgact cttttttttt cttagcgatt ggcattatca cataatgaat tatacattat
     3721 ataaagtaat gtgatttctt cgaagaatat actaaaaaat gagcaggcaa gataaacgaa
     3781 ggcaaagcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     3841 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     3901 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     3961 ggcccaccct cgtgaccacc ctgtcctggg gcgtgcagtg cttcgcccgc taccccgacc
     4021 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     4081 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     4141 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     4201 tggggcacaa gctggagtac aactacttta gcgacaacgt ctatatcacc gccgacaagc
     4261 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     4321 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     4381 acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     4441 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     4501 acaagggatc catgagagaa gttattagta ttaatggtat gttcgatttg cccgtccagg
     4561 ctagatcttt tttttgttag ttcattttgg cgtttcatct ttcattacta acttcaacgc
     4621 aaaatttttt tttttttgat ttctctttac agtcggtcaa gctggttgtc agattggtaa
     4681 tgcctgttgg gaattatatt cccttgagca cggtattaag ccggatggac atctagaaga
     4741 tggcctttca aagccgaagg gaggagaaga gggtttctcc acgtttttcc atgaaaccgg
     4801 ctacggtaag ttcgttccaa gggctattta cgtggattta gagcccaacg ttattgacga
     4861 agtccgtaac ggtccttaca aggacttgtt ccatccagaa cagttgatca gtggtaagga
     4921 ggacgcggct aataattatg caagaggcca ttacaccgtt ggtagagaaa ttttgggcga
     4981 tgttctggat aggattagaa aactggcaga ccaatgtgat gggttacaag ggttcttgtt
     5041 tacccattct cttggtggtg gtactggttc cggtctaggg tccttactgt tggaagaact
     5101 atccgctgaa tacggtaaga aatccaagct ggaatttgcc gtataccctg ctccacaagt
     5161 gtctacttct gtggttgagc cttacaacac agttttaact acacatacta cattggaaca
     5221 tgcagattgt actttcatgg tcgataatga ggctatttac gacatgtgca aaagaaactt
     5281 ggatatccca agaccaagct ttgcaaactt aaacaaccta attgctcaag tggtatcatc
     5341 tgttacagca tcattgagat tcgacggttc attaaatgta gatttgaacg aatttcaaac
     5401 caatttggtt ccatatccaa gaattcattt ccccttagtt tcatattctc cagtcttatc
     5461 caaatcaaag gcattccatg agtccaactc tgtgtcagaa attacaaacg cttgttttga
     5521 acctggtaac cagatggtca agtgtgatcc aagagatggt aaatacatgg ctacttgtct
     5581 gttatacagg ggtgatgtgg taacaagaga tgttcaaaga gctgtcgagc aggtgaaaaa
     5641 caagaagacc gtccaattgg ttgattggtg tccaactggt ttcaagatcg gtatttgcta
     5701 cgaacctcca actgccacac caaactcaca attggccact gtggataggg ccgtctgtat
     5761 gttgtcaaat accacatcca ttgctgaggc ttggaagaga atcgatagaa aattcgattt
     5821 aatgtatgcc aaacgtgctt tcgtccactg gtatgtcggt gaaggtatgg aagaaggtga
     5881 attcaccgaa gctagagaag atttggctgc tttagaaaga gattacatcg aagtgggtgc
     5941 cgactcatac gctgaggaag aggaatttta aggttcactc tcgcccccaa cctccttatc
     6001 ctttctatat ttcaagtata ttgtaaaaaa aaaaattccc cctactgtta tatctctttg
     6061 gtgatatctt atctataatc tacttattat atttctctaa tatctatgta caaaaaataa
     6121 aaaaaaaaac cattctcctt ctctaacctg catttgatga aagcttagta agtctcatcg
     6181 tagttgatgg gccaaagaca tttgtacttg tttcatcaga ttcttacatc acccatttcc
     6241 atgattattt cacggatttc attagtaatt gcatttttgt atccctagtt ccttttctcc
     6301 cgccttgccg tcgaattgaa aaaaaaaatg gaaatgagga catcttccat tgatatccag
     6361 gacaaagcaa gagtagaagt aactattagc aaggagaatt ccaaaaatag tgaacttcaa
     6421 ggtaagcaga tctatagttt caatatctga ttttctgctt ttcaaaactt aaaattcatt
     6481 agaaggtaaa acaagaaccc gtaaaattaa agttaccgat attatcaggt ttttcacccc
     6541 atgtctacta tcccatttag aaagaactat gtgttcaaaa actgggccgg tcgacctgca
     6601 gcgtacgaag cttcagctgg cggccgcgtt ctatagtgtc acctaaatcg tatgtgtatg
     6661 atacataagg ttatgtatta attgtagccg cgttctaacg acaatatgtc catatggtgc
     6721 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgcc
//
