LOCUS       sequence-75772-e        6770 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 6770)
  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 6770)
  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:mRuby2-Tub1+3'UTR::HIS3
FEATURES             Location/Qualifiers
     source          1..6770
                     /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             3789..4499
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     RBS             5984..5992
                     /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(6624..6642)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6625..6642)
                     /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 ggtgtctaag ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg
     3841 tcatggaagg ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc
     3901 cgtacatggg aactcaaacc atgaggatca aagtcatcga gggaggaccc ctgccatttg
     3961 cctttgacat tcttgccacg tcgttcatgt atggcagccg tacttttatc aagtacccga
     4021 aaggcattcc tgatttcttt aaacagtcct ttcctgaggg ttttacttgg gaaagagtta
     4081 cgagatacga agatggtgga gtcgtcaccg tcatgcagga caccagcctt gaggatggct
     4141 gtctcgttta ccacgtccaa gtcagagggg taaactttcc ctccaatggt cccgtgatgc
     4201 agaagaagac caagggttgg gagcctaata cagagatgat gtatccagca gatggtggtc
     4261 tgaggggata cactcatatg gcactgaaag ttgatggtgg tggccatctg tcttgctctt
     4321 tcgtaacaac ttacaggtca aaaaagaccg tcgggaacat caagatgccc ggtatccatg
     4381 ccgttgatca ccgcctggaa aggttagagg aaagtgacaa tgaaatgttc gtagtacaac
     4441 gcgaacacgc agttgccaag ttcgccgggc ttggtggtgg gatggacgag ctgtacaagg
     4501 gatccatgag agaagttatt agtattaatg gtatgttcga tttgcccgtc caggctagat
     4561 cttttttttg ttagttcatt ttggcgtttc atctttcatt actaacttca acgcaaaatt
     4621 tttttttttt tgatttctct ttacagtcgg tcaagctggt tgtcagattg gtaatgcctg
     4681 ttgggaatta tattcccttg agcacggtat taagccggat ggacatctag aagatggcct
     4741 ttcaaagccg aagggaggag aagagggttt ctccacgttt ttccatgaaa ccggctacgg
     4801 taagttcgtt ccaagggcta tttacgtgga tttagagccc aacgttattg acgaagtccg
     4861 taacggtcct tacaaggact tgttccatcc agaacagttg atcagtggta aggaggacgc
     4921 ggctaataat tatgcaagag gccattacac cgttggtaga gaaattttgg gcgatgttct
     4981 ggataggatt agaaaactgg cagaccaatg tgatgggtta caagggttct tgtttaccca
     5041 ttctcttggt ggtggtactg gttccggtct agggtcctta ctgttggaag aactatccgc
     5101 tgaatacggt aagaaatcca agctggaatt tgccgtatac cctgctccac aagtgtctac
     5161 ttctgtggtt gagccttaca acacagtttt aactacacat actacattgg aacatgcaga
     5221 ttgtactttc atggtcgata atgaggctat ttacgacatg tgcaaaagaa acttggatat
     5281 cccaagacca agctttgcaa acttaaacaa cctaattgct caagtggtat catctgttac
     5341 agcatcattg agattcgacg gttcattaaa tgtagatttg aacgaatttc aaaccaattt
     5401 ggttccatat ccaagaattc atttcccctt agtttcatat tctccagtct tatccaaatc
     5461 aaaggcattc catgagtcca actctgtgtc agaaattaca aacgcttgtt ttgaacctgg
     5521 taaccagatg gtcaagtgtg atccaagaga tggtaaatac atggctactt gtctgttata
     5581 caggggtgat gtggtaacaa gagatgttca aagagctgtc gagcaggtga aaaacaagaa
     5641 gaccgtccaa ttggttgatt ggtgtccaac tggtttcaag atcggtattt gctacgaacc
     5701 tccaactgcc acaccaaact cacaattggc cactgtggat agggccgtct gtatgttgtc
     5761 aaataccaca tccattgctg aggcttggaa gagaatcgat agaaaattcg atttaatgta
     5821 tgccaaacgt gctttcgtcc actggtatgt cggtgaaggt atggaagaag gtgaattcac
     5881 cgaagctaga gaagatttgg ctgctttaga aagagattac atcgaagtgg gtgccgactc
     5941 atacgctgag gaagaggaat tttaaggttc actctcgccc ccaacctcct tatcctttct
     6001 atatttcaag tatattgtaa aaaaaaaaat tccccctact gttatatctc tttggtgata
     6061 tcttatctat aatctactta ttatatttct ctaatatcta tgtacaaaaa ataaaaaaaa
     6121 aaaccattct ccttctctaa cctgcatttg atgaaagctt agtaagtctc atcgtagttg
     6181 atgggccaaa gacatttgta cttgtttcat cagattctta catcacccat ttccatgatt
     6241 atttcacgga tttcattagt aattgcattt ttgtatccct agttcctttt ctcccgcctt
     6301 gccgtcgaat tgaaaaaaaa aatggaaatg aggacatctt ccattgatat ccaggacaaa
     6361 gcaagagtag aagtaactat tagcaaggag aattccaaaa atagtgaact tcaaggtaag
     6421 cagatctata gtttcaatat ctgattttct gcttttcaaa acttaaaatt cattagaagg
     6481 taaaacaaga acccgtaaaa ttaaagttac cgatattatc aggtttttca ccccatgtct
     6541 actatcccat ttagaaagaa ctatgtgttc aaaaactggg ccggtcgacc tgcagcgtac
     6601 gaagcttcag ctggcggccg cgttctatag tgtcacctaa atcgtatgtg tatgatacat
     6661 aaggttatgt attaattgta gccgcgttct aacgacaata tgtccatatg gtgcactctc
     6721 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
//
