LOCUS       sequence-401496-        4655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CDK1-mCherry in pIVT vector for in vitro transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4655)
  AUTHORS   Davydenko O, Schultz RM, Lampson MA
  TITLE     Increased CDK1 activity determines the timing of 
            kinetochore-microtubule attachments in meiosis I.
  JOURNAL   J Cell Biol. 2013 Jul 22;202(2):221-9. doi: 10.1083/jcb.201303019. 
            Epub 2013 Jul 15.
  PUBMED    23857768
REFERENCE   2  (bases 1 to 4655)
  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: 
            "10.1083/jcb.201303019"; journalName: "J Cell Biol"; date: 
            "2013-07-22- 22"; volume: "202"; issue: "2"; pages: "221-9"
COMMENT     Sequence Label: Cdk1-mCherry
FEATURES             Location/Qualifiers
     source          1..4655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        41..59
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     41..58
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     71..90
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        71..89
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           132..174
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      193..202
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1108..1818
                     /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(1253..1271)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1521..1540
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     3'UTR           1868..1993
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2056..2073)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2056..2072)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2065..2087)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2281..2300)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2400..2422
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2460..2478)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2546..2650
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2651..2719,2720..3511)
                     /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(2869..2888)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3682..4270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4171..4190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4424..4441
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4558..4579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4594..4599,4600..4617,4618..4624)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4632..4648
                     /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     join(4637..4655,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctccaacg atttaggtga cactatagaa
       61 gagaaggaat taatacgact cactataggg agagagagag aattaccctc actaaaggga
      121 ggagaagctt gcttgttctt tttgcagaag ctcagaataa acgctcaact ttggcagatc
      181 aagcttgcat gcgccaccat ggaagactat atcaaaatag agaaaattgg agaaggtact
      241 tacggtgtgg tgtataaggg tagacacaga gtcactggcc agatagtggc catgaagaag
      301 atcagacttg aaagcgagga agaaggagtg cccagtactg caattcggga aatctctcta
      361 ttaaaagaac ttcgacatcc aaatatagtc agcctgcagg atgtgctcat gcaggactcc
      421 aggctgtatc tcatctttga gttcctgtcc atggacctca agaagtacct ggactccatc
      481 cctcctgggc agttcatgga ttcttcactc gttaagagtt acttacacca aatcctccag
      541 ggaattgtgt tttgccactc ccggcgagtt cttcacagag acttgaaacc tcaaaatcta
      601 ttgattgatg acaaaggaac aatcaaactg gctgatttcg gccttgccag agcgtttgga
      661 ataccgatac gagtgtacac acacgaggta gtgacgctgt ggtaccgatc tccagaagtg
      721 ttgctgggct cggctcgtta ctccactccg gttgacatct ggagtatagg gaccatattt
      781 gcagaactgg ccaccaagaa gccgcttttc cacggcgact cagagattga ccagctcttc
      841 aggatcttca gagctctggg cactcctaac aacgaagtgt ggccagaagt cgagtccctg
      901 caggactaca agaacacctt tcccaagtgg aagccgggga gcctcgcatc ccacgtcaag
      961 aacctggacg agaacggctt ggatttgctc tcaaaaatgc tagtctatga tcctgccaaa
     1021 cgaatctctg gcaaaatggc cctgaagcac ccgtactttg atgacttgga caatcagatt
     1081 aagaagatgg tcgactctag aggatccatg gtgagcaagg gcgaggagga taacatggcc
     1141 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     1201 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     1261 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     1321 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     1381 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     1441 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     1501 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1561 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     1621 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     1681 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     1741 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     1801 gacgagctgt acaagtaaga gctcgaattc gcccttctag tgactgacta ggatctggtt
     1861 accactaaac cagcctcaag aacacccgaa tggagtctct aagctacata ataccaactt
     1921 acactttaca aaatgttgtc ccccaaaatg tagccattcg tatctgctcc taataaaaar
     1981 aaagtttctt cacattctaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaan nccccccccc
     2041 cccaagggcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2101 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2161 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
     2221 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     2281 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     2341 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     2401 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     2461 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2521 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2581 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2641 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2701 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2761 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2821 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2881 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2941 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3001 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3061 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3121 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3181 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3241 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3301 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3361 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3421 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3481 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3541 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3601 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3661 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3721 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3781 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3841 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3901 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3961 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4021 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4081 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4141 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4201 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4261 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4321 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4381 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4441 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4501 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4561 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     4621 gttgtgtgga attgtgagcg gataacaatt tcaca
//
