LOCUS       sequence-75769-e        6781 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 6781)
  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 6781)
  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:CloverGFP-Tub1+3'UTR::HIS3
FEATURES             Location/Qualifiers
     source          1..6781
                     /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(3794..3796,3797..3799,3800..4510)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3839..3860)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     4447..4468
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             5995..6003
                     /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(6635..6653)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6636..6653)
                     /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 ggcaaagctc gagatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     3841 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtccgcggcg agggcgaggg
     3901 cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     3961 gccctggccc accctcgtga ccaccttcgg ctacggcgtg gcctgcttca gccgctaccc
     4021 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     4081 gcgcaccatc tctttcaagg acgacggtac ctacaagacc cgcgccgagg tgaagttcga
     4141 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     4201 catcctgggg cacaagctgg agtacaactt caacagccac aacgtctata tcacggccga
     4261 caagcagaag aacggcatca aggctaactt caagatccgc cacaacgttg aggacggcag
     4321 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     4381 gcccgacaac cactacctga gccatcagtc cgccctgagc aaagacccca acgagaagcg
     4441 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg attacacatg gcatggacga
     4501 gctgtacaag ggatccatga gagaagttat tagtattaat ggtatgttcg atttgcccgt
     4561 ccaggctaga tctttttttt gttagttcat tttggcgttt catctttcat tactaacttc
     4621 aacgcaaaat tttttttttt ttgatttctc tttacagtcg gtcaagctgg ttgtcagatt
     4681 ggtaatgcct gttgggaatt atattccctt gagcacggta ttaagccgga tggacatcta
     4741 gaagatggcc tttcaaagcc gaagggagga gaagagggtt tctccacgtt tttccatgaa
     4801 accggctacg gtaagttcgt tccaagggct atttacgtgg atttagagcc caacgttatt
     4861 gacgaagtcc gtaacggtcc ttacaaggac ttgttccatc cagaacagtt gatcagtggt
     4921 aaggaggacg cggctaataa ttatgcaaga ggccattaca ccgttggtag agaaattttg
     4981 ggcgatgttc tggataggat tagaaaactg gcagaccaat gtgatgggtt acaagggttc
     5041 ttgtttaccc attctcttgg tggtggtact ggttccggtc tagggtcctt actgttggaa
     5101 gaactatccg ctgaatacgg taagaaatcc aagctggaat ttgccgtata ccctgctcca
     5161 caagtgtcta cttctgtggt tgagccttac aacacagttt taactacaca tactacattg
     5221 gaacatgcag attgtacttt catggtcgat aatgaggcta tttacgacat gtgcaaaaga
     5281 aacttggata tcccaagacc aagctttgca aacttaaaca acctaattgc tcaagtggta
     5341 tcatctgtta cagcatcatt gagattcgac ggttcattaa atgtagattt gaacgaattt
     5401 caaaccaatt tggttccata tccaagaatt catttcccct tagtttcata ttctccagtc
     5461 ttatccaaat caaaggcatt ccatgagtcc aactctgtgt cagaaattac aaacgcttgt
     5521 tttgaacctg gtaaccagat ggtcaagtgt gatccaagag atggtaaata catggctact
     5581 tgtctgttat acaggggtga tgtggtaaca agagatgttc aaagagctgt cgagcaggtg
     5641 aaaaacaaga agaccgtcca attggttgat tggtgtccaa ctggtttcaa gatcggtatt
     5701 tgctacgaac ctccaactgc cacaccaaac tcacaattgg ccactgtgga tagggccgtc
     5761 tgtatgttgt caaataccac atccattgct gaggcttgga agagaatcga tagaaaattc
     5821 gatttaatgt atgccaaacg tgctttcgtc cactggtatg tcggtgaagg tatggaagaa
     5881 ggtgaattca ccgaagctag agaagatttg gctgctttag aaagagatta catcgaagtg
     5941 ggtgccgact catacgctga ggaagaggaa ttttaaggtt cactctcgcc cccaacctcc
     6001 ttatcctttc tatatttcaa gtatattgta aaaaaaaaaa ttccccctac tgttatatct
     6061 ctttggtgat atcttatcta taatctactt attatatttc tctaatatct atgtacaaaa
     6121 aataaaaaaa aaaaccattc tccttctcta acctgcattt gatgaaagct tagtaagtct
     6181 catcgtagtt gatgggccaa agacatttgt acttgtttca tcagattctt acatcaccca
     6241 tttccatgat tatttcacgg atttcattag taattgcatt tttgtatccc tagttccttt
     6301 tctcccgcct tgccgtcgaa ttgaaaaaaa aaatggaaat gaggacatct tccattgata
     6361 tccaggacaa agcaagagta gaagtaacta ttagcaagga gaattccaaa aatagtgaac
     6421 ttcaaggtaa gcagatctat agtttcaata tctgattttc tgcttttcaa aacttaaaat
     6481 tcattagaag gtaaaacaag aacccgtaaa attaaagtta ccgatattat caggtttttc
     6541 accccatgtc tactatccca tttagaaaga actatgtgtt caaaaactgg gccggtcgac
     6601 ctgcagcgta cgaagcttca gctggcggcc gcgttctata gtgtcaccta aatcgtatgt
     6661 gtatgataca taaggttatg tattaattgt agccgcgttc taacgacaat atgtccatat
     6721 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     6781 c
//
