LOCUS       sequence-237565-        7935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human E2F2 with HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7935)
  AUTHORS   Lukas J, Petersen BO, Holm K, Bartek J, Helin K
  TITLE     Deregulated expression of E2F family members induces S-phase entry 
            and overcomes p16INK4A-mediated growth suppression.
  JOURNAL   Mol Cell Biol. 1996 Mar . 16(3):1047-57.
  PUBMED    8622649
REFERENCE   2  (bases 1 to 7935)
  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: "Mol Cell 
            Biol. 1996 Mar . 16(3):1047-57."
COMMENT     Sequence Label: pCMVHA E2F2
FEATURES             Location/Qualifiers
     source          1..7935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        335..714
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        715..918
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     869..889
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     915..939
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1030..1602
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1682..1708
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3191..3210)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3256..3311
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3310..3329)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3669..3685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3669..3685)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3682..3704)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3693..3709
                     /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(3717..3723,3724..3741,3742..3747))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3762..3783
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3900..3917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4071..4659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4151..4170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4830..5621,5622..5690))
                     /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     5453..5472
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5691..5795)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5863..5881
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5919..5941)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6041..6060
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6254..6276
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6268..6285
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6269..6285
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6696..6841
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6898..7692
                     /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(6952..6971)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7562..7581
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7886..7905)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    join(7930..7935,1..42)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
       61 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
      121 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
      181 acgtcggggc ggcaagccct gccatagcca cgggccccgt gggttaggga cggcggatcg
      241 cggccctcta gagagcttgg cccattgcat acgttgtatc catatcataa tatgtacatt
      301 tatattggct catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa
      361 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      421 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      481 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      541 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      601 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      661 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      721 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      781 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      841 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      901 gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
      961 tgttttgacc tccatagaag acaccgggac cgatccagcc tccggtcgac cgatcctgag
     1021 aacttcaggg tgagtttggg gacccttgat tgttctttct ttttcgctat tgtaaaattc
     1081 atgttatatg gagggggcaa agttttcagg gtgttgttta gaatgggaag atgtcccttg
     1141 tatcaccatg gaccctcatg ataattttgt ttctttcact ttctactctg ttgacaacca
     1201 ttgtctcctc ttattttctt ttcattttct gtaacttttt cgttaaactt tagcttgcat
     1261 ttgtaacgaa tttttaaatt cacttttgtt tatttgtcag attgtaagta ctttctctaa
     1321 tcactttttt ttcaaggcaa tcagggtata ttatattgta cttcagcaca gttttagaga
     1381 acaattgtta taattaaatg ataaggtaga atatttctgc atataaattc tggctggcgt
     1441 ggaaatattc ttattggtag aaacaactac accctggtca tcatcctgcc tttctcttta
     1501 tggttacaat gatatacact gtttgagatg aggataaaat actctgagtc caaaccgggc
     1561 ccctctgcta accatgttca tgccttcttc tctttcctac agctcctggg caacgtgctg
     1621 gttgttgtgc tgtctcatca ttttggcaaa gaattcctcg acggatcaat tcaccatggc
     1681 atacccatac gatgttccag attacgctag cggatccatg ctgcaagggc cccgggcctt
     1741 ggcttcggcc gctgggcaga ccccgaaggt ggtgcccgcg atgagcccca cagagctgtg
     1801 gccatccggc ctcagcagcc cccagctctg cccagctact gctacctact acacaccgct
     1861 gtacccgcag acggcgcctc ccgcagcggc gccaggcacc tgcctcgacg ccactcccca
     1921 cggacccgag ggccaagttg tgcgatgcct gccggcaggc cggctgccgg ccaaaaggaa
     1981 gctggatctg gaggggattg ggaggcccgt cgtccctgag ttcccaaccc ccaaggggaa
     2041 gtgcatcaga gtggatggcc tccccagccc caaaaccccc aaatcccccg gggagaagac
     2101 tcggtatgac acttcgctgg ggctgctcac caagaagttc atttacctcc tgagcgagtc
     2161 agaggatggg gtcctggacc tgaactgggc cgctgaggtg ctggacgtgc agaagcggcg
     2221 catctatgac atcaccaacg tgctggaagg catccagctc atccgcaaga aggccaagaa
     2281 caacatccag tgggtaggca ggggaatgtt tgaagacccc accagacctg ggaagcagca
     2341 acagctgggg caggagctga aggagctgat gaacacggag caggccttgg accagctcat
     2401 ccagagctgc tctctgagct tcaagcacct gactgaggac aaggccaaca agaggctggc
     2461 ctatgtgact taccaggata tccgtgctgt tggcaacttt aaggagcaga cagtgattgc
     2521 cgtcaaggcc cctccgcaga cgagactgga agtgcccgac aggactgagg acaacctgca
     2581 gatatatctc aagagcaccc aagggcccat cgaagtctac ctgtgcccag aggaggtgca
     2641 ggagccggac agtccttccg aggagcctct cccctctacc tccaccctct gccccagccc
     2701 tgactctgcc cagcccagca gcagcaccga ccctagcatc atggagccca cagcatcctc
     2761 agtgccagca ccagcgccaa ccccccagca ggccccaccg cctccatccc tggtcccctt
     2821 ggaggctact gacagcctgc tggagctgcc gcacccactc ctgcagcaga ctgaggacca
     2881 gttcctgtcc ccgaccctgg cgtgcagctc ccctctgatc agcttctccc catccttgga
     2941 ccaggacgac tacctgtggg gcttggaggc gggtgagggc atcagcgatc tcttcgactc
     3001 ctacgacctt ggggacctgt tgattaattg agtggccccg cctgccccca gcagcctgcc
     3061 cccgactcta cctcctcaca gacaggctga cagccggatc cgtcgaggaa ttcactcctc
     3121 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     3181 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     3241 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     3301 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     3361 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     3421 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     3481 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     3541 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     3601 tcccagtcat agctgtccct cttctcttat ggagatccct cgacggatcg gccgcaattc
     3661 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3721 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3781 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3841 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3901 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3961 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4021 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4081 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4141 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4201 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4261 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4321 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4381 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4441 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4501 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4561 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4621 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4681 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4741 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4801 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4861 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4921 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4981 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5041 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5101 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5161 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5221 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5281 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5341 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5401 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5461 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5521 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5581 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5641 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5701 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5761 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5821 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     5881 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5941 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     6001 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     6061 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     6121 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     6181 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     6241 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttcacgct
     6301 gccgcaagca ctcagggcgc aagggctgct aaaggaagcg gaacacgtag aaagccagtc
     6361 cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa
     6421 acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact
     6481 gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg
     6541 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     6601 ggggatcaag atcctgcttc atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc
     6661 cggaagaaat atatttgcat gtctttagtt ctatgatgac acaaaccccg cccagcgtct
     6721 tgtcattggc gaattcgaac acgcagatgc agtcggggcg gcgcggtccc aggtccactt
     6781 cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg accctgcagc gacccgctta
     6841 acagcgtcaa cagcgtgccg cagatctgat caagagacag gatgaggatc gtttcgcatg
     6901 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     6961 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     7021 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     7081 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     7141 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     7201 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     7261 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     7321 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     7381 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     7441 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     7501 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     7561 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     7621 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     7681 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     7741 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     7801 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     7861 accccgggag atgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     7921 gcgctatgac ggcaa
//
