LOCUS       sequence-206242-        4848 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  mCherry-eDHFR-Cdc42Q61L in pIVT vector for in vitro transcription. 
            Note that Cdc42 in this plasmid keeps its CAAX domain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4848)
  AUTHORS   Akera T, Chmatal L, Trimm E, Yang K, Aonbangkhen C, Chenoweth DM, 
            Janke C, Schultz RM, Lampson MA
  TITLE     Spindle asymmetry drives non-Mendelian chromosome segregation.
  JOURNAL   Science. 2017 Nov 3;358(6363):668-672. doi: 10.1126/science.aan0092.
  PUBMED    29097549
REFERENCE   2  (bases 1 to 4848)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2017-11-3- 3"; volume: "358"; issue: 
            "6363"; pages: "668-672"
COMMENT     Sequence Label: mCherry-eDHFR-Cdc42Q61L
FEATURES             Location/Qualifiers
     source          1..4848
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           62..104
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             165..872
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(310..328)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     578..597
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             891..1364
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             join(1371..1550,1551..1553,1554..1946)
                     /codon_start=1
                     /product="constitutively active mutant of human Rho family 
                     small GTPase Cdc42"
                     /label=Cdc42 (Q61L)
                     /translation="MQTIKCVVVGDGAVGKTCLLISYTTNKFPSEYVPTVFDNYAVTVM
                     IGGEPYTLGLFDTAGLEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHH
                     CPKTPFLLVGTQIDLRDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQK
                     GLKNVFDEAILAALEPPEPKKSRRCVLL"
     3'UTR           1996..2121
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2179..2196)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2179..2195)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2188..2210)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2404..2423)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2523..2545
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2583..2601)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2669..2773
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2774..2842,2843..3634)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELDEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2992..3011)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3805..4393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4294..4313
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4547..4564
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4681..4702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4717..4722,4723..4740,4741..4747)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4755..4771
                     /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)."
     primer_bind     4760..4782
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4779..4795
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4779..4795
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4819..4837
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4819..4836
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 taatacgact cactataggg agagagagag aattaccctc actaaaggga ggagaagctt
       61 gcttgttctt tttgcagaag ctcagaataa acgctcaact ttggcagatc aagcttgcat
      121 gcctgcaggt cgactctaga ggatccggcg cgccaccggt caccatggtg agcaagggcg
      181 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
      241 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
      301 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
      361 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
      421 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
      481 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
      541 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
      601 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
      661 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
      721 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
      781 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
      841 actccaccgg cggcatggac gagctgtata aggccggcct ggagtctaga atcagtctga
      901 ttgcggcgtt agcggtagat cgcgttatcg gcatggaaaa cgccatgccg tggaacctgc
      961 ctgccgatct cgcctggttt aaacgcaaca ccttaaataa acccgtgatt atgggccgcc
     1021 atacctggga atcaatcggt cgtccgttgc caggacgcaa aaatattatc ctcagcagtc
     1081 aaccgggtac ggacgatcgc gtaacgtggg tgaagtcggt ggatgaagcc atcgcggcgt
     1141 gtggtgacgt accagaaatc atggtgattg gcggcggtcg cgtttatgaa cagttcttgc
     1201 caaaagcgca aaaactgtat ctgacgcata tcgacgcaga agtggaaggc gacacccatt
     1261 tcccggatta cgagccggat gactgggaat cggtattcag cgaattccac gatgctgatg
     1321 cgcagaactc tcacagctat tgctttgaga ttctggagcg gcgggagctc atgcagacaa
     1381 ttaagtgtgt tgttgtgggc gatggtgctg ttggtaaaac atgtctcctg atatcctaca
     1441 caacaaacaa atttccatcg gaatatgtac cgactgtttt tgacaactat gcagtcacag
     1501 ttatgattgg tggagaacca tatactcttg gactttttga tactgcaggg ctagaggatt
     1561 atgacagatt acgaccgctg agttatccac aaacagatgt atttctagtc tgtttttcag
     1621 tggtctctcc atcttcattt gaaaacgtga aagaaaagtg ggtgcctgag ataactcacc
     1681 actgtccaaa gactcctttc ttgcttgttg ggactcaaat tgatctcaga gatgacccct
     1741 ctactattga gaaacttgcc aagaacaaac agaagcctat cactccagag actgctgaaa
     1801 agctggcccg tgacctgaag gctgtcaagt atgtggagtg ttctgcactt acacagaaag
     1861 gcctaaagaa tgtatttgac gaagcaatat tggctgccct ggagcctcca gaaccgaaga
     1921 agagccgcag gtgtgtgctg ctatgagagc tcgaattcgc ccttctagtg actgactagg
     1981 atctggttac cactaaacca gcctcaagaa cacccgaatg gagtctctaa gctacataat
     2041 accaacttac actttacaaa atgttgtccc ccaaaatgta gccattcgta tctgctccta
     2101 ataaaaagaa agtttcttca cattctaaaa aaaaaaaaaa aaaaaaaaaa aaaacccccc
     2161 ccccccaagg gcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     2221 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     2281 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     2341 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     2401 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     2461 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2521 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     2581 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     2641 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2701 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2761 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2821 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2881 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2941 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3001 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3061 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3121 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3181 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3241 tgtaactcgc cttgatcgtt gggaaccgga gctggatgaa gccataccaa acgacgagcg
     3301 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3361 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3421 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3481 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3541 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3601 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3661 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3721 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3781 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3841 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3901 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3961 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4021 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4081 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4141 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4201 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4261 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4321 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4381 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4441 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4501 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4561 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     4621 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     4681 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     4741 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     4801 ttacgccaag ctccaacgat ttaggtgaca ctatagaaga gaaggaat
//
