LOCUS Exported 6822 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses mouse Egr2 in mammalian cells. ACCESSION . VERSION . KEYWORDS pcDNA 3.1(-) mouse Egr2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6822) AUTHORS Salotti J, Sakchaisri K, Tourtellotte WG, Johnson PF TITLE An Arf-Egr-C/EBPbeta pathway linked to ras-induced senescence and cancer. JOURNAL Mol Cell Biol. 2015 Mar;35(5):866-83. doi: 10.1128/MCB.01489-14. Epub 2014 Dec 22. PUBMED 25535333 REFERENCE 2 (bases 1 to 6822) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6822 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 12..391 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 392..595 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 546..566 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 640..659 /label=T7 /note="T7 promoter, forward primer" promoter 640..658 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2190..2207) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2196..2420 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2466..2894 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2553..2572) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2763..2784 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2903..2923) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2908..3237 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3088..3223 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3150..3169 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3304..4098 /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(3358..3377) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3968..3987 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4272..4393 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4309..4328) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4363..4382 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4442..4458) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4442..4458) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4455..4477) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4466..4482 /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(4490..4520) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4535..4556 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4673..4690) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4844..5432) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4924..4943) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5603..6463) /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 6226..6245 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6464..6568) /gene="bla" /label=AmpR promoter primer_bind complement(6643..6662) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt 61 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg 121 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac 181 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt 241 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa 301 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta 361 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg 421 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg 481 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc 541 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg 601 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga 661 gacccaagct ggctagcgtt taaacgggcc ctctagactc gagcggccgc cactgtgctg 721 gatatctgca gaattcgcgc caccatgatg accgccaagg ccgtagacaa aatcccagta 781 actctcagtg gttttatgca ccagctgcct gacagcctct acccggtgga agacctcgcc 841 gcctcgtcgg tgaccatctt ccccaatggt gaactgggag gcccctttga ccagatgaac 901 ggagtggcgg gagatggcat gatcaacatt gacatgactg gagagaagag acccctggat 961 ctcccgtatc cgagtagctt cgctcccatc tctgcaccta gaaaccagac cttcacttac 1021 atgggcaaat tctccattga cccacagtac cctggtgcca gctgctatcc agaaggtatc 1081 atcaatattg tgagtgcggg catcttgcaa ggggtcaccc ctccagcttc aaccacagcc 1141 tcctccagcg tcacctccgc ctcccccaac ccactggcca cgggacccct gggtgtgtgt 1201 accatgtccc agactcagcc tgaactggac cacctctact ctccgccacc acctcctcct 1261 ccttattcgg gctgtacagg agatctctac caggatcctt cagcattctt atcgccgcca 1321 tccaccactt ccacctcctc tctggcctac cagccacctc cttcctaccc atcccccaag 1381 ccagctatgg acccaggtct cattcctatg atcccagact atcctggatt ttttccatct 1441 ccgtgccaga gagatccaca cggtgctgct ggcccagatc gaaagccgtt tccctgtcct 1501 ctggactccc tgcgagtgcc ccctccactc acgccactct ctaccatccg taattttact 1561 ctggggggtc ccggtgctgg agtcacggga ccaggagcaa gtggaggtgg tgagggacct 1621 cggctgcctg gcagtgggtc tgcagcagtg actgccaccc cttataatcc gcaccacctg 1681 ccattgcggc ccatcctgcg acctcgaaag taccctaaca ggcccagcaa aacgccagtg 1741 cacgaaaggc cctatccctg cccagcagaa ggttgtgata ggaggttctc acgctctgat 1801 gagctgacca ggcacatccg aatccacacg ggccacaagc ccttccagtg tcggatctgc 1861 atgcgaaact tcagccgaag tgaccacctt actactcaca tccgaaccca caccggggag 1921 aagccctttg cctgtgacta ttgtggccgc aagtttgcca ggagtgacga aaggaagcgc 1981 cacaccaaga tccaccttcg gcagaaggaa cggaagagca gtgctccctc tgcacctcca 2041 tctgcccagt cttcagcctc tggtcctggg ggctcgcagg ccgggggcag cctgtgcggt 2101 aacagcgcca ttggaggacc actggcctcc tgcacctctc gaaccaggac accgtgagtc 2161 gacaagctta agtttaaacc gctgatcagc ctcgactgtg ccttctagtt gccagccatc 2221 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct 2281 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg 2341 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg 2401 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta 2461 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 2521 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 2581 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 2641 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 2701 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 2761 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 2821 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 2881 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 2941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 3001 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 3061 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 3121 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc 3181 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag 3241 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt 3301 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 3361 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 3421 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 3481 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 3541 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 3601 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 3661 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 3721 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 3781 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 3841 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 3901 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 3961 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 4021 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4081 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 4141 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 4201 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 4261 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 4321 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 4381 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 4441 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 4501 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 4561 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 4621 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 4681 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 4741 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 4801 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 4861 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 4921 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 4981 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 5041 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 5101 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 5161 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 5221 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 5281 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 5341 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 5401 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 5461 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 5521 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 5581 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 5641 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 5701 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 5761 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 5821 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 5881 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 5941 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 6001 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 6061 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 6121 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 6181 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 6241 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 6301 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 6361 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 6421 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 6481 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 6541 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg 6601 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc 6661 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc 6721 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct 6781 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc ag //