LOCUS       sequence-152182-        7897 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of human E2F1 with HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7897)
  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 7897)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 E2F1
FEATURES             Location/Qualifiers
     source          1..7897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        145..524
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        525..728
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     679..699
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     725..749
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          840..1412
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1492..1518
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2963..2982)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3028..3083
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3082..3101)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3441..3457)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3441..3457)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3454..3476)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3465..3481
                     /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(3489..3495,3496..3513,3514..3519))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3534..3555
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3672..3689)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3843..4431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3923..3942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4602..5393,5394..5462))
                     /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     5225..5244
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5463..5567)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5635..5653
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5691..5713)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5813..5832
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6026..6048
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6040..6057
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6041..6057
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6468..6613
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6670..7464
                     /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(6724..6743)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7334..7353
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7658..7677)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7702..7749
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ggcaagccct gccatagcca cgggccccgt gggttaggga cggcggatcg cggccctcta
       61 gagagcttgg cccattgcat acgttgtatc catatcataa tatgtacatt tatattggct
      121 catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
      181 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      241 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      301 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      361 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      421 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      481 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      541 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      601 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      661 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      721 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
      781 tccatagaag acaccgggac cgatccagcc tccggtcgac cgatcctgag aacttcaggg
      841 tgagtttggg gacccttgat tgttctttct ttttcgctat tgtaaaattc atgttatatg
      901 gagggggcaa agttttcagg gtgttgttta gaatgggaag atgtcccttg tatcaccatg
      961 gaccctcatg ataattttgt ttctttcact ttctactctg ttgacaacca ttgtctcctc
     1021 ttattttctt ttcattttct gtaacttttt cgttaaactt tagcttgcat ttgtaacgaa
     1081 tttttaaatt cacttttgtt tatttgtcag attgtaagta ctttctctaa tcactttttt
     1141 ttcaaggcaa tcagggtata ttatattgta cttcagcaca gttttagaga acaattgtta
     1201 taattaaatg ataaggtaga atatttctgc atataaattc tggctggcgt ggaaatattc
     1261 ttattggtag aaacaactac accctggtca tcatcctgcc tttctcttta tggttacaat
     1321 gatatacact gtttgagatg aggataaaat actctgagtc caaaccgggc ccctctgcta
     1381 accatgttca tgccttcttc tctttcctac agctcctggg caacgtgctg gttgttgtgc
     1441 tgtctcatca ttttggcaaa gaattcctcg acggatcaat tcaccatggc atacccatac
     1501 gatgttccag attacgctag cggatccgcc atggccttgg ccggggcccc tgcgggcggc
     1561 ccatgcgcgc cggcgctgga ggccctgctc ggggccggcg cgctgcggct gctcgactcc
     1621 tcgcagatcg tcatcatctc cgccgcgcag gacgccagcg ccccgccggc tcccaccggc
     1681 cccgcggcgc ccgccgccgg cccctgcgac cctgacctgc tgctcttcgc cacaccgcag
     1741 gcgccccggc ccacacccag tgcgccgcgg cccgcgctcg gccgcccgcc ggtgaagcgg
     1801 aggctggacc tggaaactga ccatcagtac ctggccgaga gcagtgggcc agctcggggc
     1861 agaggccgcc atccaggaaa aggtgtgaaa tccccggggg agaagtcacg ctatgagacc
     1921 tcactgaatc tgaccaccaa gcgcttcctg gagctgctga gccactcggc tgacggtgtc
     1981 gtcgacctga actgggctgc cgaggtgctg aaggtgcaga agcggcgcat ctatgacatc
     2041 accaacgtcc ttgagggcat ccagctcatt gccaagaagt ccaagaacca catccagtgg
     2101 ctgggcagcc acaccacagt gggcgtcggc ggacggcttg aggggttgac ccaggacctc
     2161 cgacagctgc aggagagcga gcagcagctg gaccacctga tgaatatctg tactacgcag
     2221 ctgcgcctgc tctccgagga cactgacagc cagcgcctgg cctacgtgac gtgtcaggac
     2281 cttcgtagca ttgcagaccc tgcagagcag atggttatgg tgatcaaagc ccctcctgag
     2341 acccagctcc aagccgtgga ctcttcggag aactttcaga tctcccttaa gagcaaacaa
     2401 ggcccgatcg atgttttcct gtgccctgag gagaccgtag gtgggatcag ccctgggaag
     2461 accccatccc aggaggtcac ttctgaggag gagaacaggg ccactgactc tgccaccata
     2521 gtgtcaccac caccatcatc tcccccctca tccctcacca cagatcccag ccagtctcta
     2581 ctcagcctgg agcaaaaacc gctgttgtcc cggatgggca gcctgcgggc tcccgtggac
     2641 gaggaccgcc tgtccccgct ggtggcggcc gactcgctcc tggagcatgt gcgggaggac
     2701 ttctccggcc tcctccctga ggagttcatc agcctttccc caccccacga ggccctcgac
     2761 taccacttcg gcctcgagga gggcgagggc atcagagacc tcttcgactg tgactttggg
     2821 gacctcaccc ccctggattt ctgacagggc tggatccact agttctagga tccgtcgagg
     2881 aattcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
     2941 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
     3001 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     3061 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     3121 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     3181 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     3241 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     3301 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     3361 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacggat
     3421 cggccgcaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3481 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3541 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3601 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3661 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3721 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3781 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3841 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3901 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3961 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4021 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4081 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4141 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4201 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4261 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4321 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4381 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4441 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4501 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4561 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4621 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4681 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4741 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4801 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4861 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4921 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4981 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5041 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5101 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5161 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5221 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5281 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5341 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5401 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5461 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5521 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5581 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     5641 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5701 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     5761 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     5821 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     5881 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     5941 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     6001 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
     6061 aagcttcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag cggaacacgt
     6121 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     6181 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     6241 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     6301 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     6361 tctgatggcg caggggatca agatcctgct tcatccccgt ggcccgttgc tcgcgtttgc
     6421 tggcggtgtc cccggaagaa atatatttgc atgtctttag ttctatgatg acacaaaccc
     6481 cgcccagcgt cttgtcattg gcgaattcga acacgcagat gcagtcgggg cggcgcggtc
     6541 ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag cgaccctgca
     6601 gcgacccgct taacagcgtc aacagcgtgc cgcagatctg atcaagagac aggatgagga
     6661 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6721 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6781 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6841 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6901 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6961 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     7021 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     7081 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     7141 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     7201 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     7261 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     7321 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     7381 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     7441 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     7501 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     7561 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     7621 agttcttcgc ccaccccggg agatggggga ggctaactga aacacggaag gagacaatac
     7681 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     7741 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     7801 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     7861 cgggtgaagg cccagggctc gcagccaacg tcggggc
//
