LOCUS Exported 8845 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Constitutive ETV4 ORF, puromycin selection. ACCESSION . VERSION . KEYWORDS pLX_TRC311 ETV4-S SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8845) AUTHORS Wang B, Krall EB, Aguirre AJ, Kim M, Widlund HR, Doshi MB, Sicinska E, Sulahian R, Goodale A, Cowley GS, Piccioni F, Doench JG, Root DE, Hahn WC TITLE ATXN1L, CIC, and ETS Transcription Factors Modulate Sensitivity to MAPK Pathway Inhibition. JOURNAL Cell Rep. 2017 Feb 7;18(6):1543-1557. doi: 10.1016/j.celrep.2017.01.031. PUBMED 28178529 REFERENCE 2 (bases 1 to 8845) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8845 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 98..1276 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 329..1267 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1224..1244 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" protein_bind 1296..1320 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(1942..1966) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 1974..2015 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" misc_feature 2066..2654 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2119..2139) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2537..2548) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 2726..2959 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 3037..3158 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3074..3093) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3128..3147 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 3198..3333 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(3353..3372) /label=T7 /note="T7 promoter, forward primer" promoter complement(3354..3372) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3382..3399) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3382..3398) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3391..3413) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3540..3995 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3627..3646) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3837..3858 /label=F1ori-F /note="F1 origin, forward primer" promoter 4021..4125 /gene="bla" /label=AmpR promoter CDS 4126..4986 /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 complement(4344..4363) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5157..5745 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5646..5665 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5899..5916 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6033..6054 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6069..6099 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6107..6123 /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)." primer_bind 6112..6134 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6131..6147 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6131..6147 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 6166..6186 /label=T3 /note="T3 promoter, forward primer" promoter 6168..6186 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 6214..6440 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 6441..6621 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 6668..6793 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 7286..7519 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7704..7748 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 7979..8189 /label=SV40 promoter /note="SV40 early promoter" promoter 8001..8197 /label=SV40 promoter /note="SV40 early promoter" rep_origin 8048..8183 /label=SV40 ori /note="SV40 origin of replication" CDS 8219..8614 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" misc_feature 8688..8805 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 ggctcgaggc ctgcaggtgc aaagatggat aaagttttaa acagagagga atctttgcag 61 ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc 121 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg 181 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct 241 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt 301 tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg 361 cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc tgcagtacgt 421 gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta 481 aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt 541 gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta 601 aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg 661 ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc 721 gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg 781 ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc 841 cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc 901 gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc 961 gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga 1021 ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac 1081 gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt 1141 ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt 1201 tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc 1261 atttcaggtg tcgtgaggct agcatcgatt gatcaacaag tttgtacaaa aaagttggca 1321 tgtacctcca cacagagggc ttctctgggc cctctccagg tgacggggcc atgggctatg 1381 gctatgagaa acctctgcga ccattcccag atgatgtctg cgttgtccct gagaaatttg 1441 aaggagacat caagcaggaa ggggtcggtg catttcgaga ggggccgccc taccagcgcc 1501 ggggtgccct gcagctgtgg caatttctgg tggccttgct ggatgaccca acaaatgccc 1561 atttcattgc ctggacgggc cggggaatgg agttcaagct cattgagcct gaggaggtcg 1621 ccaggctctg gggcatccag aagaaccggc cagccatgaa ttacgacaag ctgagccgct 1681 cgctccgata ctattatgag aaaggcatca tgcagaaggt ggctggtgag cgttacgtgt 1741 acaagtttgt gtgtgagccc gaggccctct tctctttggc cttcccggac aatcagcgtc 1801 cagctctcaa ggctgagttt gaccggcctg tcagtgagga ggacacagtc cctttgtccc 1861 acttggatga gagccccgcc tacctcccag agctggctgg ccccgcccag ccatttggcc 1921 ccaagggtgg ctactcttac tacccaactt tcttgtacaa agtggttgat atcggtaagc 1981 ctatccctaa ccctctcctc ggtctcgatt ctacgtagta atgaactagt accggttaag 2041 tcgacaatca acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt 2101 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc 2161 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg 2221 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac 2281 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc 2341 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc 2401 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa 2461 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc 2521 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc 2581 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg 2641 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc 2701 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat 2761 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct 2821 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta 2881 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca 2941 gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta 3001 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat 3061 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 3121 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc 3181 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 3241 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat 3301 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag 3361 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc 3421 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag 3481 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga 3541 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 3601 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 3661 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 3721 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 3781 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 3841 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 3901 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 3961 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg 4021 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 4081 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 4141 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 4201 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 4261 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 4321 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg 4381 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 4441 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 4501 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 4561 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 4621 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca 4681 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta 4741 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct 4801 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca 4861 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag 4921 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat 4981 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt 5041 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 5101 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 5161 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 5221 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 5281 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 5341 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 5401 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 5461 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 5521 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 5581 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 5641 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 5701 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 5761 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 5821 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 5881 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc 5941 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc 6001 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca 6061 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa 6121 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac 6181 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt 6241 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg 6301 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg 6361 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt 6421 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct 6481 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 6541 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 6601 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc 6661 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg 6721 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg 6781 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt 6841 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct 6901 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact 6961 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac 7021 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt 7081 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga 7141 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata 7201 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc 7261 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag 7321 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg 7381 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc 7441 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc 7501 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg 7561 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga 7621 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg 7681 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg 7741 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa 7801 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata 7861 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac 7921 ccatgcattg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 7981 aggcagaagt atgcaaagca tgcgtctcaa ttagtcagca accatagtcc cgcccctaac 8041 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 8101 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 8161 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctttctagag gtaccaccat 8221 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa 8281 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg 8341 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt 8401 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa 8461 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca 8521 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg 8581 tgaattgctg ccctctggtt atgtgtggga gggcctgcag ctgcagtagt aaggcgcgcc 8641 gttaacgaat tctagatctt gagacaaatg gcagtattca tccacaattt taaaagaaaa 8701 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata 8761 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg 8821 gacagcagag atccactttg gcgcc //