LOCUS       sequence-473585-        6726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of TFDP1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6726)
  AUTHORS   Clijsters L, Hoencamp C, Calis JJA, Marzio A, Handgraaf SM, Cuitino 
            MC, Rosenberg BR, Leone G, Pagano M
  TITLE     Cyclin F Controls Cell-Cycle Transcriptional Outputs by Directing 
            the Degradation of the Three Activator E2Fs.
  JOURNAL   Mol Cell. 2019 Jun 20;74(6):1264-1277.e7. doi: 
            10.1016/j.molcel.2019.04.010. Epub 2019 May 23.
  PUBMED    31130363
REFERENCE   2  (bases 1 to 6726)
  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: 
            "10.1016/j.molcel.2019.04.010"; journalName: "Mol Cell"; date: 
            "2019-06-20- 20"; volume: "74"; issue: "6"; pages: "1264-1277.e7"
COMMENT     Sequence Label: pcDNA3 2xFLAG-2xSTREP TFDP1
FEATURES             Location/Qualifiers
     source          1..6726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      304..892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     793..812
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1046..1063
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1180..1201
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1216..1221,1222..1239,1240..1246)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1254..1270
                     /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     1259..1281
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1278..1294
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1278..1294
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1343..1464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1354..1373)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1408..1427
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1638..2432)
                     /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(1749..1768)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2359..2378
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2499..2828)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2513..2648
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2567..2586)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2813..2833
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2842..3270)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2952..2973)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3164..3183
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3316..3540
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3529..3546
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4779..4802)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(4839..4862)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(4878..4901)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(4917..4940)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4981..5000)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4982..5000)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5045..5248)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5074..5094)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5249..5628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5800..5819
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5894..5998
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5999..6067,6068..6726,1..133)
                     /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(6217..6236)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
       61 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
      121 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
      181 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
      241 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
      301 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
      361 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
      421 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
      481 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
      541 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
      601 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
      661 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
      721 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      781 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      841 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      901 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      961 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     1021 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     1081 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     1141 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     1201 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     1261 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga
     1321 ggtcgacggt atacagacat gataagatac attgatgagt ttggacaaac cacaactaga
     1381 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1441 attataagct gcaataaaca agttggggtg ggcgaagaac tccagcatga gatccccgcg
     1501 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     1561 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     1621 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     1681 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     1741 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     1801 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     1861 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg
     1921 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     1981 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     2041 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     2101 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     2161 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     2221 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg
     2281 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     2341 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     2401 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     2461 tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca
     2521 aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc ggcctctgca
     2581 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     2641 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     2701 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     2761 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     2821 tgacacacat tccacagaat taattcgcgt taaatttttg ttaaatcagc tcatttttta
     2881 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     2941 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3001 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3061 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3121 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3181 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3241 ccgcgcttaa tgcgccgcta cagggcgcgt ggggataccc cctagagccc cagctggttc
     3301 tttccgcctc agaagccata gagcccaccg catccccagc atgcctgcta ttgtcttccc
     3361 aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca
     3421 gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca
     3481 gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag
     3541 tcgaggctga tcagcgggtt taaacgggcc ctctagactc gagcggccgc tcagtcgtcc
     3601 tcgtcattct cgttgaagtc atcgtcctcc tcgtcgtcct ccccgacgta ggacaccgga
     3661 gtctccaccc tggagccgct gtactgagac ccattggagc ttgtggccag catcccatct
     3721 gcaccgttgg tcaggtcact ggcagagaac cttgtgccgt tagacgtgga acctgccgtc
     3781 gtgatgaaca cgcctccaac agttccctga gccatttctg tcacgtatgg ctccagagcc
     3841 ttggggacca gacttctggc cattttaagg tcttcggcag agcagctccc cgactccagc
     3901 ccgcaagcca tgcccatccg cttcagcact tctatgtcat cgtggatttc aaatgtgttg
     3961 tcaaaattaa acagatactc aaatttgtca ttggagatgc tgcagtcgat gaccgtcttc
     4021 ttgctggtgt tgacgatgat gaagggcagg tggatgactg agttgggtgg cggtggccgg
     4081 ctggcctgct gctccgcatg ccggtttctc tgcaccaggt tcttgaaggc aatttgctgt
     4141 agaataagtt cttgaagttg agactgtttc tgttttattc tttcaagtct cctctgtctt
     4201 tccacctcta agttctgaca ttcctgagcc gagttggtgg gcagaccaat ccacttgatc
     4261 tccttcttct ccttggagat gatgttcatg gccattagca cgtttaaggc atcgtagacg
     4321 cgccgtctta tgtttttctg gtcataagct gactcgtttg gtaagatgtg gttgtcggca
     4381 gcactgaact ccgcaaccag ctcgtctgcc acttcgttgt aggaagtggt acctttcctc
     4441 tgcaccatgg cagaccaagg tgaggagtcg gaaggctggt tctgagaggc aaagtgagtg
     4501 ctgggggtgt gtgggcttcc taccaccagg gtgtttgacg ctgccggtct ctgaggcgta
     4561 ccaattacca cttgctgggc aatgttgaca ttggactgtc caaaggtttt tggcaagagc
     4621 tgcttcccga gcgggttgac ggtggagggg tgaacggcca cgagggacac cacgcctttc
     4681 ccgggactaa ggttctggtc tatgaagacc ttgagttctc cgttggcttc aattagaccg
     4741 gcatcttttg cgaattccac cacactggat ccagctcctt tctcgaactg cgggtggctc
     4801 cagcttcctc ctccgctgcc tccgccgctt cctcctccct tctcgaactg agggtggctc
     4861 cagctggcgg ccgacccttt atcatcatca tctttataat ccatagatcc tggtaccttg
     4921 tcatcgtcgt ccttgtagtc catggtaagc ttaagtttaa acgctagcca gcttgggtct
     4981 ccctatagtg agtcgtatta atttcgataa gccagtaagc agtgggttct ctagttagcc
     5041 agagagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     5101 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     5161 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     5221 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     5281 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     5341 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     5401 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     5461 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     5521 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     5581 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc
     5641 tggcccgtac atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat
     5701 tgtcggtcaa gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct
     5761 ccaacacaca agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac
     5821 tgagagtgca ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt
     5881 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5941 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6001 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6061 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6121 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6181 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6241 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6301 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6361 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6421 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6481 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6541 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6601 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6661 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6721 cggtat
//
