LOCUS       sequence-172568-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of cyclin D1 T286A mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4377)
  AUTHORS   Matsushime H, Roussel MF, Ashmun RA, Sherr CJ
  TITLE     Colony-stimulating factor 1 regulates novel cyclins during the G1 
            phase of the cell cycle.
  JOURNAL   Cell. 1991 May 17;65(4):701-13.
  PUBMED    1827757
REFERENCE   2  (bases 1 to 4377)
  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: "Cell."; 
            date: "1991-05-17"; volume: "65(4)"; pages: "701-13"
COMMENT     Sequence Label: pFLEX-m-cyclin D1 -T286A mutant
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..24
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    975..1109
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(999..1018)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1053..1072
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1252..1274)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1360..1500
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1415..1434
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1515..1532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1686..2274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1766..1785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2445..3236,3237..3305))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3068..3087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3306..3410)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3478..3496
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3566..3586)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(3571..3900,3901..3915,3916..4184)
                     /label=SR-alpha promoter
                     /note="hybrid promoter consisting of the SV40 early 
                     promoter plus part of the long terminal repeat of human 
                     T-lymphotrophic virus 1"
     rep_origin      3751..3886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     LTR             3916..4184
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          4228..4324
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
ORIGIN
        1 gactacaagg acgacgatga caagtctcga gcaattgaat tcggccgcgc catggaacac
       61 cagctcctgt gctgcgaagt ggagaccatc cgccgcgcgt accctgacac caatctcctc
      121 aacgaccggg tgctgcgagc catgctcaag acggaggaga cctgtgcgcc ctccgtatct
      181 tacttcaagt gcgtgcagaa ggagattgtg ccatccatgc ggaaaatcgt ggccacctgg
      241 atgctggagg tctgtgagga gcagaagtgc gaagaggagg tcttcccgct ggccatgaac
      301 tacctggacc gcttcctgtc cctggagccc ctgaagaaga gccgcctgca gctgctgggg
      361 gccacctgca tgttcgtggc ctctaagatg aaggagacca ttcccttgac tgccgagaaa
      421 ttgtgcatct acactgacaa ctctatccgg cccgaggagc tgctgcaaat ggaactgctt
      481 ctggtgaaca agctcaagtg gaacctggcc gccatgactc cccacgattt catcgaacac
      541 ttcctctcca aaatgccaga ggcggatgag aacaagcaga ccatccgcaa gcatgcacag
      601 acctttgtgg ccctctgtgc cacagatgtg aagttcattt ccaacccacc ctccatggta
      661 gctgctggga gcgtggtggc tgcgatgcaa ggcctgaacc tgggcagccc caacaacttc
      721 ctctcctgct accgcacaac gcactttctt tccagagtca tcaagtgtga cccggactgc
      781 ctccgtgcct gccaggaaca gattgaagcc cttctggagt caagcctgcg ccaggcccag
      841 cagaacgtcg accccaaggc cactgaggag gagggggaag tggaggaaga ggctggtctg
      901 gcctgcgcgc ccaccgacgt gcgagatgtg gacatctgag gatcccgcaa gccgaattcc
      961 tgattgattg accggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     1021 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     1081 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     1141 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     1201 tggctgatta tgatccggct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     1261 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     1321 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     1381 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     1441 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     1501 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1561 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1621 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1681 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1741 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1801 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1861 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1921 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1981 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2041 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2101 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     2161 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2221 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2281 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2341 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2401 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2461 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2521 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2581 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2641 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2701 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2761 gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2821 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2881 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2941 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3001 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3061 acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3121 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3181 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3241 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3301 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3361 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3421 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     3481 aggcgtatca cgaggccctt tcgtcttcaa gaattggtcg accaattctc atgtttgaca
     3541 gcttatcatc gataagcttg gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3601 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3661 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3721 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3781 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3841 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3901 aagctcctcg atcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc
     3961 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact
     4021 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc
     4081 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa
     4141 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct cctcgatcga
     4201 ggaactgaaa aaccagaaag ttaactggta agtttagtct ttttgtcttt tatttcaggt
     4261 cccggatccg gtggtggtgc aaatcaaaga actgctcctc agtggatgtt gcctttactt
     4321 ctaggcctgt acggaagtgt tacttctgct ctaaaagctg ctctagccct cgccatg
//
