LOCUS       sequence-75748-e        7178 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 7178)
  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 7178)
  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::HPH
FEATURES             Location/Qualifiers
     source          1..7178
                     /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"
     gene            complement(2326..3901)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     terminator      complement(2326..2523)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(2529..3557)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(2529..3557)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(3558..3901)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        4009..4195
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             4197..4907
                     /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             6392..6400
                     /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(7032..7050)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7033..7050)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7131..7150)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
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 tagtggatct gatatcatcg atgaattcga
     2281 gctcgttttc gacactggat ggcggcgtta gtatcgaatc gacagcagta tagcgaccag
     2341 cattcacata cgattgacgc atgatattac tttctgcgca cttaacttcg catctgggca
     2401 gatgatgtcg aggcgaaaaa aaatataaat cacgctaaca tttgattaaa atagaacaac
     2461 tacaatataa aaaaactata caaatgacaa gttcttgaaa acaagaatct ttttattgtc
     2521 agtactgatt attcctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg
     2581 agtacttcta cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc
     2641 ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca
     2701 tcatcgaaat tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata
     2761 tacgcccgga gccgcggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc
     2821 tgctgctcca tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg
     2881 gaatccccga acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc
     2941 aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg
     3001 gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc
     3061 acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta
     3121 gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg
     3181 gccgcagcga tcgcatccat ggcctccgcg accggctgca gaacagcggg cagttcggtt
     3241 tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg agatgcaata ggtcaggctc
     3301 tcgctgaatt ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc
     3361 cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat
     3421 ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc
     3481 aggtcggaga cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt
     3541 tcaggctttt tacccatggt tgtttatgtt cggatgtgat gtgagaactg tatcctagca
     3601 agattttaaa aggaagtata tgaaagaaga acctcagtgg caaatcctaa ccttttatat
     3661 ttctctacag gggcgcggcg tggggacaat tcaacgcgtc tgtgagggga gcgtttccct
     3721 gctcgcaggt ctgcagcgag gagccgtaat ttttgcttcg cgccgtgcgg ccatcaaaat
     3781 gtatggatgc aaatgattat acatggggat gtatgggcta aatgtacggg cgacagtcac
     3841 atcatgcccc tgagctgcgc acgtcaagac tgtcaaggag ggtattctgg gcctccatgt
     3901 cgctggccgg gtgacccggc ggggacgagg caagctaaac agatctggcg cgccttaatt
     3961 aaccactaaa gggaacaaaa gctggtaccc tattactctt ggcctcctct agtacactct
     4021 atattttttt atgcctcggt aatgattttc attttttttt tccacctagc ggatgactct
     4081 ttttttttct tagcgattgg cattatcaca taatgaatta tacattatat aaagtaatgt
     4141 gatttcttcg aagaatatac taaaaaatga gcaggcaaga taaacgaagg caaagcatgg
     4201 tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc atggaaggtt
     4261 cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga aggcaatccg tacatgggaa
     4321 ctcaaaccat gaggatcaaa gtcatcgagg gaggacccct gccatttgcc tttgacattc
     4381 ttgccacgtc gttcatgtat ggcagccgta cttttatcaa gtacccgaaa ggcattcctg
     4441 atttctttaa acagtccttt cctgagggtt ttacttggga aagagttacg agatacgaag
     4501 atggtggagt cgtcaccgtc atgcaggaca ccagccttga ggatggctgt ctcgtttacc
     4561 acgtccaagt cagaggggta aactttccct ccaatggtcc cgtgatgcag aagaagacca
     4621 agggttggga gcctaataca gagatgatgt atccagcaga tggtggtctg aggggataca
     4681 ctcatatggc actgaaagtt gatggtggtg gccatctgtc ttgctctttc gtaacaactt
     4741 acaggtcaaa aaagaccgtc gggaacatca agatgcccgg tatccatgcc gttgatcacc
     4801 gcctggaaag gttagaggaa agtgacaatg aaatgttcgt agtacaacgc gaacacgcag
     4861 ttgccaagtt cgccgggctt ggtggtggga tggacgagct gtacaaggga tccatgagag
     4921 aagttattag tattaatggt atgttcgatt tgcccgtcca ggctagatct tttttttgtt
     4981 agttcatttt ggcgtttcat ctttcattac taacttcaac gcaaaatttt tttttttttg
     5041 atttctcttt acagtcggtc aagctggttg tcagattggt aatgcctgtt gggaattata
     5101 ttcccttgag cacggtatta agccggatgg acatctagaa gatggccttt caaagccgaa
     5161 gggaggagaa gagggtttct ccacgttttt ccatgaaacc ggctacggta agttcgttcc
     5221 aagggctatt tacgtggatt tagagcccaa cgttattgac gaagtccgta acggtcctta
     5281 caaggacttg ttccatccag aacagttgat cagtggtaag gaggacgcgg ctaataatta
     5341 tgcaagaggc cattacaccg ttggtagaga aattttgggc gatgttctgg ataggattag
     5401 aaaactggca gaccaatgtg atgggttaca agggttcttg tttacccatt ctcttggtgg
     5461 tggtactggt tccggtctag ggtccttact gttggaagaa ctatccgctg aatacggtaa
     5521 gaaatccaag ctggaatttg ccgtataccc tgctccacaa gtgtctactt ctgtggttga
     5581 gccttacaac acagttttaa ctacacatac tacattggaa catgcagatt gtactttcat
     5641 ggtcgataat gaggctattt acgacatgtg caaaagaaac ttggatatcc caagaccaag
     5701 ctttgcaaac ttaaacaacc taattgctca agtggtatca tctgttacag catcattgag
     5761 attcgacggt tcattaaatg tagatttgaa cgaatttcaa accaatttgg ttccatatcc
     5821 aagaattcat ttccccttag tttcatattc tccagtctta tccaaatcaa aggcattcca
     5881 tgagtccaac tctgtgtcag aaattacaaa cgcttgtttt gaacctggta accagatggt
     5941 caagtgtgat ccaagagatg gtaaatacat ggctacttgt ctgttataca ggggtgatgt
     6001 ggtaacaaga gatgttcaaa gagctgtcga gcaggtgaaa aacaagaaga ccgtccaatt
     6061 ggttgattgg tgtccaactg gtttcaagat cggtatttgc tacgaacctc caactgccac
     6121 accaaactca caattggcca ctgtggatag ggccgtctgt atgttgtcaa ataccacatc
     6181 cattgctgag gcttggaaga gaatcgatag aaaattcgat ttaatgtatg ccaaacgtgc
     6241 tttcgtccac tggtatgtcg gtgaaggtat ggaagaaggt gaattcaccg aagctagaga
     6301 agatttggct gctttagaaa gagattacat cgaagtgggt gccgactcat acgctgagga
     6361 agaggaattt taaggttcac tctcgccccc aacctcctta tcctttctat atttcaagta
     6421 tattgtaaaa aaaaaaattc cccctactgt tatatctctt tggtgatatc ttatctataa
     6481 tctacttatt atatttctct aatatctatg tacaaaaaat aaaaaaaaaa accattctcc
     6541 ttctctaacc tgcatttgat gaaagcttag taagtctcat cgtagttgat gggccaaaga
     6601 catttgtact tgtttcatca gattcttaca tcacccattt ccatgattat ttcacggatt
     6661 tcattagtaa ttgcattttt gtatccctag ttccttttct cccgccttgc cgtcgaattg
     6721 aaaaaaaaaa tggaaatgag gacatcttcc attgatatcc aggacaaagc aagagtagaa
     6781 gtaactatta gcaaggagaa ttccaaaaat agtgaacttc aaggtaagca gatctatagt
     6841 ttcaatatct gattttctgc ttttcaaaac ttaaaattca ttagaaggta aaacaagaac
     6901 ccgtaaaatt aaagttaccg atattatcag gtttttcacc ccatgtctac tatcccattt
     6961 agaaagaact atgtgttcaa aaactgggcc ggtcgacctg cagcgtacga agcttcagct
     7021 ggcggccgcg ttctatagtg tcacctaaat cgtatgtgta tgatacataa ggttatgtat
     7081 taattgtagc cgcgttctaa cgacaatatg tccatatggt gcactctcag tacaatctgc
     7141 tctgatgccg catagttaag ccagccccga cacccgcc
//
