LOCUS       sequence-473586-        6723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of TFDP2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6723)
  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 6723)
  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 TFDP2
FEATURES             Location/Qualifiers
     source          1..6723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     189..209
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(218..646)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(328..349)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     540..559
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    692..916
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     905..922
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2152..2175)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(2212..2235)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(2251..2274)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2290..2313)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2354..2373)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2355..2373)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2418..2621)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2447..2467)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2622..3001
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3173..3192
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3267..3371
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3372..3440,3441..4232)
                     /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(3590..3609)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4403..4991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4892..4911
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5145..5162
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5279..5300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5315..5320,5321..5338,5339..5345)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5353..5369
                     /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     5358..5380
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5377..5393
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5377..5393
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5442..5563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5453..5472)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5507..5526
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5737..6531)
                     /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(5848..5867)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6458..6477
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6598..6723,1..204))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6612..6723,1..24)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6666..6685)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
       61 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
      121 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      181 ctttccacac cctaactgac acacattcca cagaattaat tcgcgttaaa tttttgttaa
      241 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      301 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      361 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      421 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      481 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      541 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      601 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg atacccccta
      661 gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc cccagcatgc
      721 ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata
      781 gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg
      841 ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg
      901 caactagaag gcacagtcga ggctgatcag cgggtttaaa cgggccctct agactcgagc
      961 ggccgcttat tctggggagg aggaatcctc ctcatcatct tcctcatctt catcattgaa
     1021 cgaacagggg gtctcgcctc gggactcgga gcagtgagag gccgcactgc tggactggtg
     1081 actgtttggg gccaggaact gcccagttgc taaggccact tctgcatcca agcataaccc
     1141 ttggtttaca cttgactggg gtaaggtggc accagtggtc aggtctaaat ttgaaactga
     1201 ttgggtagag ttcagaagta gtccctgatt taaccaagaa ggtcctgtgg agatatctgt
     1261 gatataacct tctaaagcct ttggcaccag ggatttcgca agtttcagat cctccagaga
     1321 gcatttgcct gactccaggc caaacgacat tcccatccgc tttagtactt ctatgtcatc
     1381 atggatctca aaggtgttgt caaaattgaa aagatactca aacttgtcac tggagatgct
     1441 gcaatctatg actgtttttc tgcttgtatt gatgattatg aatggcagct gaatggtaga
     1501 gttcagagcc ggcgggccct ggttttgctg ctcattttgt cgatttctct gtaccaggtt
     1561 tttgaaagcg atttgctgta ggagaagttc ttgcagctgg gcccgcttct gctttatccg
     1621 ttctatccgc ctctgcttct ctatctccag attctgacat tcctgagcag aattggtagg
     1681 caggccaatc cacttgattt cttttttttc ctttgaaatt atgttcattg ccattagcac
     1741 atttaaagca tcataaactc ttcgcctaat gttcttctga tcataagcct gcgaatcagc
     1801 agccaaatgg ttatttgaat tggtgaactc tgacaccagc tcatcagcga cttcattgta
     1861 cgatgttgta ccttttcgtt gaactttctc acacactttc attgaaaagt gtctcaagcc
     1921 tttcccattt ttatctcctt ttttgcttcg tttactttct gaaaaatcag agtctataaa
     1981 ttttctagcc cgttttctat caccagggac ccagccagta gcttctgcta tgtgtgtctg
     2041 agtaaccatt gctggtgcag gggtatatgg actcccaatc agaacacttc ctgaactggt
     2101 tagtctctgt ggtgtgctta taatgaattc caccacactg gatccagctc ctttctcgaa
     2161 ctgcgggtgg ctccagcttc ctcctccgct gcctccgccg cttcctcctc ccttctcgaa
     2221 ctgagggtgg ctccagctgg cggccgaccc tttatcatca tcatctttat aatccataga
     2281 tcctggtacc ttgtcatcgt cgtccttgta gtccatggta agcttaagtt taaacgctag
     2341 ccagcttggg tctccctata gtgagtcgta ttaatttcga taagccagta agcagtgggt
     2401 tctctagtta gccagagagc tctgcttata tagacctccc accgtacacg cctaccgccc
     2461 atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc
     2521 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
     2581 cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact
     2641 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     2701 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     2761 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     2821 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     2881 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     2941 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt
     3001 caacgcgtat atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga
     3061 ttcttcatgc aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac
     3121 tactcagcga cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca
     3181 gagcagattg tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt
     3241 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3301 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3361 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3421 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3481 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3541 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3601 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3661 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3721 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3781 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3841 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3901 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3961 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4021 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4081 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4141 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4201 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4261 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4321 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4381 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4441 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4501 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4561 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4621 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4681 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4741 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4801 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4861 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4921 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4981 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5041 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5101 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5161 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5221 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5281 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5341 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     5401 acgccaagct ctagctagag gtcgacggta tacagacatg ataagataca ttgatgagtt
     5461 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     5521 tattgcttta tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact
     5581 ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga
     5641 agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg
     5701 tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     5761 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     5821 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     5881 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     5941 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     6001 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     6061 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     6121 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     6181 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     6241 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     6301 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt
     6361 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     6421 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     6481 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     6541 tcctcatcct gtctcttgat cagatccgaa aatggatata caagctcccg ggagcttttt
     6601 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggcaga
     6661 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     6721 gga
//
