LOCUS       sequence-148919-        6320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Centromeric plasmid to construct episomal C-terminal fusions of 
            genes with yeast-adapted TagRFP-T and HIS3 selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6320)
  AUTHORS   Malcova I, Farkasovsky M, Senohrabkova L, Vasicova P, Hasek J
  TITLE     New integrative modules for multicolor-protein labeling and 
            live-cell imaging in Saccharomyces cerevisiae.
  JOURNAL   FEMS Yeast Res. 2016 Mar 17. pii: fow027.
  PUBMED    26994102
REFERENCE   2  (bases 1 to 6320)
  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"; journalName: "FEMS Yeast 
            Res. 2016 Mar 17. pii: fow027."
COMMENT     Sequence Label: pIM23
FEATURES             Location/Qualifiers
     source          1..6320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        317..503
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             504..1166
                     /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"
     rep_origin      complement(1426..1881)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1563..1584)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1775..1794
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2011..2033
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2025..2042
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2026..2042
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2052..2071
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2052..2070
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2090..2336)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2302..2320
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3129..3145)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3129..3145)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(3146..3495)
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     complement(3546..3566)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3546..3564)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3585..3601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3585..3601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3598..3620)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3609..3625
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3633..3639,3640..3657,3658..3663))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3816..3833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3987..4575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4067..4086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4746..5537,5538..5606))
                     /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     5369..5388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5607..5711)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5748..6251
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6293..6311
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgcgtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
      361 tttttttttt cccctagcgg atgactcttt ttttttctta gcgattggca ttatcacata
      421 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc
      481 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa
      541 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
      601 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
      661 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
      721 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
      781 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctaggg gccgtgcgtg
      841 gagtaaaaag gtttggatca ggatttgcgc ctttggatga ggcactttcc agagcggtgg
      901 tagatctttc gaacaggccg tacgcagttg tcgaacttgg tttgcaaagg gagaaagtag
      961 gagatctctc ttgcgagatg atcccgcatt ttcttgaaag ctttgcagag gctagcagaa
     1021 ttaccctcca cgttgattgt ctgcgaggca agaatgatca tcaccgtagt gagagtgcgt
     1081 tcaaggctct tgcggttgcc ataagagaag ccacctcgcc caatggtacc aacgatgttc
     1141 cctccaccaa aggtgttctt atgtagtgac accgattatt taaagctgca gcatacgata
     1201 tatatacatg tgtatatatg tatacctatg aatgtcagta agtatgtata cgaacagtat
     1261 gatactgaag atgacaaggt aatgcatcat tctatacgtg tcattctgaa cgaggcgcgc
     1321 tttccttttt tctttttgct ttttcttttt ttttctcttg aactcgacgg atctatgcgg
     1381 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaaacgtta
     1441 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1501 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1561 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1621 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     1681 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     1741 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     1801 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     1861 atgcgccgct acagggcgcg tcgcgccatt cgccattcag gctgcgcaac tgttgggaag
     1921 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     1981 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     2041 gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccggccac aaattaaagc
     2101 cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt ataatgttac atgcgtacac
     2161 gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat aacgttctta atactaacat
     2221 aactataaaa aaataaatag ggacctagac ttcaggttgt ctaactcctt ccttttcggt
     2281 tagagcggat gtggggggag ggcgtgaatg taagcgtgac ataactaatt acatgactcg
     2341 accagtttta cttatacaat tcgtccatac cgttcaactt atgacctaac ttggaaggca
     2401 aatcgcagta tcttgcaaca gcgacttcgt gttgttcaac atatgtttcc ttgtctgctt
     2461 ctttaattct ttccaatcta tgatcgacgt agtatacacc aggcatcttc aagtttttag
     2521 ctggcttttt agatctgtaa gtggtcttga aattgcaaat taagtgacca ccaccaacta
     2581 atttcaatgc catgtcagtt cgaccttcta aaccaccatc ggcaggatac aacatttcgg
     2641 tgtttgcttc ccaacccaaa gttttctttt gcattacagg accgttactt gggaagttta
     2701 cacctctgat cttaacgtta tagattaaac aaccgtcttg caaggatgta tcttgtgtgg
     2761 cagtcaaaac accaccgtct tcatatgtag tgactctttc ccaagtgaaa ccttcaggga
     2821 atgattgctt aaagaaatct ggtatacctt gagtatgatt gatgaaggtt cttgaaccgt
     2881 acataaaaga tgtagctaag atatcgaagg caaaaggcaa tggaccacct tctacaactt
     2941 taattctcat ggtttgtgta ccttcatatg gtttaccttc accttcagag gtacacttaa
     3001 aatggtggtt atttacagta ccttccatat acaacttcat gtgcatgttt tccttaatca
     3061 attcttcacc cttagatacc atgtcgacgg tatcgataag cttgatatcg aattcctgca
     3121 gcccggggga tccactagtt ctagagtatg gatgggggta atagaattgt atctatgtat
     3181 ctgacgaccc tgtattacac gaaggaaata gtagatgaaa agacaagaga gcaagaaaaa
     3241 ggaaaaactt cctttctaac agacgcttta cttaacctta tatatatctt atttttttca
     3301 tcgagcgtgt tcaattggac aaggtgccac cttttcgaca cttcggttat tatgttacac
     3361 agttttcatg aggatggcgc cttgactaac ttaattagtc atttgccaac caccacagtt
     3421 ccccaatatc gacagcttca cgtgccattt gccattttgc gatcatgtga cctcaagaga
     3481 aaaagcaaaa aataatgaga gctaagggat tcgaaccctt gcatccggag ctccagcttt
     3541 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     3601 gtgtgaaatt gttatccgct cacaattcca cacaacatag gagccggaag cataaagtgt
     3661 aaagcctggg gtgcctaatg agtgaggtaa ctcacattaa ttgcgttgcg ctcactgccc
     3721 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3781 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3841 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3901 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3961 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4021 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4081 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4141 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4201 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4261 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4321 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4381 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4441 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4501 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4561 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4621 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4681 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4741 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4801 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4861 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4921 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4981 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5041 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5101 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5161 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5221 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5281 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5341 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5401 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5461 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5521 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5581 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5641 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5701 ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt ccttttcatc acgtgctata
     5761 aaaataatta taatttaaat tttttaatat aaatatataa attaaaaata gaaagtaaaa
     5821 aaagaaatta aagaaaaaat agtttttgtt ttccgaagat gtaaaagact ctagggggat
     5881 cgccaacaaa tactaccttt tatcttgctc ttcctgctct caggtattaa tgccgaattg
     5941 tttcatcttg tctgtgtaga agaccacaca cgaaaatcct gtgattttac attttactta
     6001 tcgttaatcg aatgtatatc tatttaatct gcttttcttg tctaataaat atatatgtaa
     6061 agtacgcttt ttgttgaaat tttttaaacc tttgtttatt tttttttctt cattccgtaa
     6121 ctcttctacc ttctttattt actttctaaa atccaaatac aaaacataaa aataaataaa
     6181 cacagagtaa attcccaaat tattccatca ttaaaagata cgaggcgcgt gtaagttaca
     6241 ggcaagcgat ccgtcctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6301 tatcacgagg ccctttcgtc
//
