LOCUS       sequence-189831-        6737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human E2F1 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6737)
  AUTHORS   Hoog G, Zarrizi R, von Stedingk K, Jonsson K, Alvarado-Kristensson M
  TITLE     Nuclear localization of gamma-tubulin affects E2F transcriptional 
            activity and S-phase progression.
  JOURNAL   FASEB J. 2011 Nov;25(11):3815-27. doi: 10.1096/fj.11-187484. Epub 
            2011 Jul 25.
  PUBMED    21788450
REFERENCE   2  (bases 1 to 6737)
  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: 
            "10.1096/fj.11-187484"; journalName: "FASEB J"; date: "2011-11"; 
            volume: "25"; issue: "11"; pages: "3815-27"
COMMENT     Sequence Label: pET21 E2F1
FEATURES             Location/Qualifiers
     source          1..6737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(974..2056)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2018..2037
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2057..2134)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2340..2359
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2443..2462
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2443..2461
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2462..2486
                     /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)."
     RBS             2501..2523
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2530..2562
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             3911..3928
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3981..3999)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3995..4042
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4079..4534
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4166..4185)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4376..4397
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4560..4664
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4665..4733,4734..5525)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4883..4902)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5696..6284
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6185..6204
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6438..6455
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6470..6609
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6535..6554)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6695..6717
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(6711..6737,1..165))
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
       61 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
      121 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
      181 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
      241 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
      301 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
      361 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
      421 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
      481 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
      541 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
      601 caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg
      661 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
      721 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
      781 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
      841 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
      901 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
      961 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1021 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     1081 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     1141 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     1201 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     1261 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     1321 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     1381 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     1441 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     1501 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     1561 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     1621 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     1681 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     1741 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     1801 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     1861 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     1921 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     1981 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     2041 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     2101 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     2161 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     2221 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     2281 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     2341 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     2401 atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag
     2461 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg
     2521 agatatacca tggctagcat gactggtgga cagcaaatgg gtcggatccg aattcaccat
     2581 ggccttggcc ggggcccctg cgggcggccc atgcgcgccg gcgctggagg ccctgctcgg
     2641 ggccggcgcg ctgcggctgc tcgactcctc gcagatcgtc atcatctccg ccgcgcagga
     2701 cgccagcgcc ccgccggctc ccaccggccc cgcggcgccc gccgccggcc cctgcgaccc
     2761 tgacctgctg ctcttcgcca caccgcaggc gccccggccc acacccagtg cgccgcggcc
     2821 cgcgctcggc cgcccgccgg tgaagcggag gctggacctg gaaactgacc atcagtacct
     2881 ggccgagagc agtgggccag ctcggggcag aggccgccat ccaggaaaag gtgtgaaatc
     2941 cccgggggag aagtcacgct atgagacctc actgaatctg accaccaagc gcttcctgga
     3001 gctgctgagc cactcggctg acggtgtcgt cgacctgaac tgggctgccg aggtgctgaa
     3061 ggtgcagaag cggcgcatct atgacatcac caacgtcctt gagggcatcc agctcattgc
     3121 caagaagtcc aagaaccaca tccagtggct gggcagccac accacagtgg gcgtcggcgg
     3181 acggcttgag gggttgaccc aggacctccg acagctgcag gagagcgagc agcagctgga
     3241 ccacctgatg aatatctgta ctacgcagct gcgcctgctc tccgaggaca ctgacagcca
     3301 gcgcctggcc tacgtgacgt gtcaggacct tcgtagcatt gcagaccctg cagagcagat
     3361 ggttatggtg atcaaagccc ctcctgagac ccagctccaa gccgtggact cttcggagaa
     3421 ctttcagatc tcccttaaga gcaaacaagg cccgatcgat gttttcctgt gccctgagga
     3481 gaccgtaggt gggatcagcc ctgggaagac cccatcccag gaggtcactt ctgaggagga
     3541 gaacagggcc actgactctg ccaccatagt gtcaccacca ccatcatctc ccccctcatc
     3601 cctcaccaca gatcccagcc agtctctact cagcctggag caagaaccgc tgttgtcccg
     3661 gatgggcagc ctgcgggctc ccgtggacga ggaccgcctg tccccgctgg tggcggccga
     3721 ctcgctcctg gagcatgtgc gggaggactt ctccggcctc ctccctgagg agttcatcag
     3781 cctttcccca ccccacgagg ccctcgacta ccacttcggc ctcgaggagg gcgagggcat
     3841 cagagacctc ttcgactgtg actttgggga cctcaccccc ctggatttca agcttgcggc
     3901 cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     3961 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     4021 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
     4081 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4141 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4201 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4261 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4321 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4381 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4441 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4501 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     4561 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4621 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4681 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4741 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4801 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4861 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4921 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4981 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5041 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5101 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5161 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     5221 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5281 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5341 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5401 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5461 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5521 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5581 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5641 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5701 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5761 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5821 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     5881 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5941 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6001 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6061 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6121 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6181 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6241 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6301 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6361 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6421 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     6481 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     6541 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     6601 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     6661 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     6721 tcaccgtcat caccgaa
//
