LOCUS       sequence-206243-        4267 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  mCherry-eDHFR in pIVT vector for in vitro transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4267)
  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 4267)
  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
FEATURES             Location/Qualifiers
     source          1..4267
                     /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"
     3'UTR           1414..1539
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(1598..1615)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1598..1614)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1607..1629)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1823..1842)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1942..1964
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2002..2020)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2088..2192
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2193..2261,2262..3053)
                     /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(2411..2430)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3224..3812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3713..3732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3966..3983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4100..4121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4136..4141,4142..4159,4160..4166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4174..4190
                     /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     4179..4201
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4198..4214
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4198..4214
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4238..4256
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4238..4255
                     /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 gaattcgccc
     1381 ttctagtgac tgactaggat ctggttacca ctaaaccagc ctcaagaaca cccgaatgga
     1441 gtctctaagc tacataatac caacttacac tttacaaaat gttgtccccc aaaatgtagc
     1501 cattcgtatc tgctcctaat aaaaagaaag tttcttcaca ttctaaaaaa aaaaaaaaaa
     1561 aaaaaaaaaa aacccccccc cccccaaggg cgaattcact ggccgtcgtt ttacaacgtc
     1621 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1681 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1741 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     1801 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     1861 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     1921 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     1981 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2041 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2101 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2161 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2221 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2281 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2341 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2401 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2461 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2521 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2581 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2641 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctggatgaag
     2701 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2761 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2821 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2881 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2941 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3001 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3061 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3121 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3181 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3241 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3301 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3361 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3421 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3481 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3541 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3601 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3661 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3721 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3781 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3841 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3901 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3961 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     4021 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     4081 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     4141 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     4201 gaaacagcta tgaccatgat tacgccaagc tccaacgatt taggtgacac tatagaagag
     4261 aaggaat
//
