LOCUS       sequence-245679-        8270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human E2F5 with HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8270)
  TITLE     K Helin plasmid
REFERENCE   2  (bases 1 to 8270)
  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"
COMMENT     Sequence Label: pCMVHA-E2F5
FEATURES             Location/Qualifiers
     source          1..8270
                     /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     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          696..1268
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1348..1374
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      1382..1391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1545..1562
                     /codon_start=1
                     /product="recognition and cleavage site for Genenase(TM) I,
                     a subtilisin BPN' variant engineered for increased 
                     substrate specificity (Carter and Wells, 1987)"
                     /label=Genenase(TM) I site
                     /translation="PGAAHY"
     primer_bind     complement(3192..3211)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3257..3312
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3311..3330)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3670..3686)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3670..3686)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3683..3705)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3694..3710
                     /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(3718..3724,3725..3742,3743..3748))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3763..3784
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3901..3918)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4072..4660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4152..4171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4831..5622,5623..5691))
                     /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     5454..5473
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5692..5796)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5864..5882
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5920..5942)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6042..6061
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6255..6277
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6269..6286
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6270..6286
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6697..6842
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6899..7693
                     /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(6953..6972)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7563..7582
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7887..7906)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7931..7978
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
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 agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg gtcgaccgat cctgagaact tcagggtgag tttggggacc cttgattgtt
      721 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
      781 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
      841 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
      901 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
      961 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
     1021 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
     1081 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc
     1141 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
     1201 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt
     1261 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat
     1321 tcctcgacgg atcaattcac catggcatac ccatacgatg ttccagatta cgctagcgga
     1381 tccatggcgg cggcagagcc cgcgagctcg ggccagcagg cgccggcagg gcaggggcag
     1441 ggccagcggc cgccgccgca gcctccgcag gcgcaagccc cgcagccgcc cccgccgccg
     1501 cagctcgggg gcgcgggggg cggcagcagc aggcacgaga agagcctggg gctgctcact
     1561 accaagttcg tgtcgctgct gcaggaggcc aaggacggcg ttctggatct caaagcggct
     1621 gctgatactt tggctgtgag gcaaaaaagg agaatttatg atatcaccaa tgtcttagag
     1681 ggaattgact tgattgaaaa aaagtcaaaa aacagtatcc agtggaaagg tgtaggtgct
     1741 ggctgtaata ctaaagaagt catagataga ttaagatatc ttaaagctga aattgaagat
     1801 ctagaactga aggaaagaga acttgatcag cagaagttgt ggctacagca aagcatcaaa
     1861 aatgtgatgg acgattccat taataataga ttttcctatg taactcatga agacatctgt
     1921 aattgcttta atggtgatac acttttggcc attcaggcac cttctggtac acaactggag
     1981 gtacccattc cagaaatggg tcagaatgga caaaagaaat accagatcaa tctaaagagt
     2041 cattcaggac ctatccatgt gctgcttata aataaagagt cgagttcatc taagcccgtg
     2101 gtttttcctg ttcccccacc tgatgacctc acacagcctt cctcccagtc cttgactcca
     2161 gtgactccac agaaatccag catggcaact caaaatctgc ctgagcaaca tgtctctgaa
     2221 agaagccagg ctctgcagca gacatcagct acagatatat cttcagcagg atctattagt
     2281 ggagatatca ttgatgagtt aatgtcttct gacgtgtttc ctctcttaag gctttctcct
     2341 accccggcag atgactacaa ctttaattta gatgataacg aaggagtttg tgatctgttt
     2401 gatgtccaga tactaaatta ttagattcca tggaaacttg ggactgttat ctacctctaa
     2461 ctgtgtaaca ttttagactt cttaataacc taaatattta aaataatgaa tgtaacacct
     2521 tttttagttc actgattctg aagtgttctt ccctaatact ttctttactt cacaaaactt
     2581 caaccataaa aacaaagggc tctgattgct ttaggggata agtgatttaa tatccacaaa
     2641 cgtccccact cccaaaagta actatattct ggatttcaac ttttcttcta attgtgaatc
     2701 cttctgtttt ttcttcttaa ggaggaaagt taaaggacac tacaggtcat caaaaacaag
     2761 ttggccaagg actcattact tgtcttatat ttttactgcc actaaactgc ctgtatttct
     2821 gtatgtcctt ctatccaaac agacgttcac tgccacttgt aaagtgaagg atgtaaacga
     2881 ggatatataa ctgtttcagt gaacagattt tgtgaagtgc cttctgtttt agcactttaa
     2941 gtttatcaca ttttgttgac ttctgacatt ccactttcct aggttatagg aaagatctgt
     3001 ttatgtagtt tgtttttaaa atgtgccaat gcctgtacat taacaagatt tttaaaaata
     3061 aaattgtata aaaaaaaaaa aaaaaaaaaa aatctaggat ccgtcgagga attcactcct
     3121 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     3181 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     3241 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     3301 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     3361 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     3421 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     3481 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     3541 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     3601 ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacggatc ggccgcaatt
     3661 cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3721 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3781 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3841 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3901 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3961 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4021 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4081 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4141 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4201 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4261 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4321 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4381 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4441 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4501 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4561 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4621 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4681 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4741 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4801 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4861 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4921 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4981 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5041 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5101 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5161 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5221 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5281 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5341 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5401 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5461 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5521 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5581 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5641 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5701 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5761 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5821 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5881 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     5941 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     6001 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     6061 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     6121 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6181 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     6241 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agcttcacgc
     6301 tgccgcaagc actcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt
     6361 ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa
     6421 aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac
     6481 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
     6541 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc
     6601 aggggatcaa gatcctgctt catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc
     6661 ccggaagaaa tatatttgca tgtctttagt tctatgatga cacaaacccc gcccagcgtc
     6721 ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc caggtccact
     6781 tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag cgacccgctt
     6841 aacagcgtca acagcgtgcc gcagatctga tcaagagaca ggatgaggat cgtttcgcat
     6901 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     6961 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     7021 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     7081 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     7141 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     7201 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     7261 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     7321 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     7381 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     7441 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     7501 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     7561 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     7621 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     7681 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     7741 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     7801 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     7861 caccccggga gatgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     7921 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     7981 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     8041 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     8101 ccagggctcg cagccaacgt cggggcggca agccctgcca tagccacggg ccccgtgggt
     8161 tagggacggc ggatcgcggc cctctagaga gcttggccca ttgcatacgt tgtatccata
     8221 tcataatatg tacatttata ttggctcatg tccaacatta ccgccatgtt
//
