LOCUS       sequence-238545-        6344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Cyclin D1 protein with a Myc tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6344)
  AUTHORS   Lessard F, Igelmann S, Trahan C, Huot G, Saint-Germain E, Mignacca 
            L, Del Toro N, Lopes-Paciencia S, Le Calve B, Montero M, 
            Deschenes-Simard X, Bury M, Moiseeva O, Rowell MC, Zorca CE, 
            Zenklusen D, Brakier-Gingras L, Bourdeau V, Oeffinger M, Ferbeyre G
  TITLE     Senescence-associated ribosome biogenesis defects contributes to 
            cell cycle arrest through the Rb pathway.
  JOURNAL   Nat Cell Biol. 2018 Jul;20(7):789-799. doi: 
            10.1038/s41556-018-0127-y. Epub 2018 Jun 25.
  PUBMED    29941930
REFERENCE   2  (bases 1 to 6344)
  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: 
            "10.1038/s41556-018-0127-y"; journalName: "Nat Cell Biol"; date: 
            "2018-07"; volume: "20"; issue: "7"; pages: "789-799"
COMMENT     Sequence Label: pcDNA3-Myc-CyclinD1(WT)
FEATURES             Location/Qualifiers
     source          1..6344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             688..717
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      1316..1325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1663..1681)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1664..1681)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1701..1718)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1707..1931
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1977..2405
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2064..2083)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2274..2295
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2414..2434)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2419..2748
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2599..2734
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2661..2680
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2815..3609
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2869..2888)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3479..3498
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3783..3904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3820..3839)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3874..3893
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3953..3969)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3953..3969)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3966..3988)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3977..3993
                     /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(4001..4007,4008..4025,4026..4031))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4046..4067
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4184..4201)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4355..4943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4435..4454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5114..5905,5906..5974))
                     /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     5737..5756
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5975..6079)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6154..6173)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac catggcagaa caaaaactta tttctgaaga agatctggaa
      721 caccagctcc tgtgctgcga agtggaaacc atccgccgcg cgtaccccga tgccaacctc
      781 ctcaacgacc gggtgctgcg ggccatgctg aaggcggagg agacctgcgc gccctcggtg
      841 tcctacttca aatgtgtgca gaaggaggtc ctgccgtcca tgcggaagat cgtcgccacc
      901 tggatgctgg aggtctgcga ggaacagaag tgcgaggagg aggtcttccc gctggccatg
      961 aactacctgg accgcttcct gtcgctggag cccgtgaaaa agagccgcct gcagctgctg
     1021 ggggccactt gcatgttcgt ggcctctaag atgaaggaga ccatccccct gacggccgag
     1081 aagctgtgca tctacaccga caactccatc cggcccgagg agctgctgca aatggagctg
     1141 ctcctggtga acaagctcaa gtggaacctg gccgcaatga ccccgcacga tttcattgaa
     1201 cacttcctct ccaaaatgcc agaggcggag gagaacaaac agatcatccg caaacacgcg
     1261 cagaccttcg ttgccctctg tgccacagat gtgaagttca tttccaatcc gccctccatg
     1321 gtggcagcgg ggagcgtggt ggccgcagtg caaggcctga acctgaggag ccccaacaac
     1381 ttcctgtcct actaccgcct cacacgcttc ctctccagag tgatcaagtg tgacccggac
     1441 tgcctccggg cctgccagga gcagatcgaa gccctgctgg agtcaagcct gcgccaggcc
     1501 cagcagaaca tggaccccaa ggccgccgag gaggaggaag aggaggagga ggaggtggac
     1561 ctggcttgca cacccaccga cgtgcgggac gtggacatct gagaattctg cagatatcca
     1621 tcacactggc ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa
     1681 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1741 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1801 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1861 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     1921 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     1981 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2041 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2101 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2161 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2221 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2281 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2341 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2401 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2461 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2521 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2581 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2641 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     2701 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     2761 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2821 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2881 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2941 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3001 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3061 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3121 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3181 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3241 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3301 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3361 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3421 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3481 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3541 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3601 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3661 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3721 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3781 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3841 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3901 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     3961 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4021 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4081 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4141 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4201 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4261 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4321 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4381 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4441 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4501 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4561 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4621 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4681 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4741 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4801 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4861 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4921 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4981 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5041 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5101 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5161 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5221 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5281 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5341 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5401 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5461 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5521 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5581 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5641 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5701 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5761 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5821 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5881 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5941 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6001 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6061 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     6121 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6181 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     6241 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     6301 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
