LOCUS Exported 7119 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION gene expression of EGFP-TIA1. ACCESSION . VERSION . KEYWORDS pFRT_TO_eGFP_TIA1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7119) AUTHORS Meyer C, Garzia A, Mazzola M, Gerstberger S, Molina H, Tuschl T TITLE The TIA1 RNA-Binding Protein Family Regulates EIF2AK2-Mediated Stress Response and Cell Cycle Progression. JOURNAL Mol Cell. 2018 Feb 15;69(4):622-635.e6. doi: 10.1016/j.molcel.2018.01.011. PUBMED 29429924 REFERENCE 2 (bases 1 to 7119) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7119 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 820..838 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 841..859 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 869..893 /label=LNCX /note="Human CMV promoter, forward primer" CDS 990..1706 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 990..1706 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1035..1056) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1296..1315) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1643..1664 /label=EGFP-C /note="EGFP, forward primer" protein_bind 1713..1737 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2915..2939) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2992..3033 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(3071..3088) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3077..3301 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(3434..3453) /label=F1ori-R /note="F1 origin, reverse primer" protein_bind 3585..3632 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS 3640..4662 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " polyA_signal 4792..4913 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4829..4848) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4883..4902 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4962..4978) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4962..4978) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4975..4997) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4986..5002 /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(5010..5040) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5055..5076 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5193..5210) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5364..5952) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5444..5463) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6123..6983) /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 6746..6765 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6984..7088) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc 841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga 901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga 961 ctctagcgtt taaacttaag cttggtacca tggtgagcaa gggcgaggag ctgttcaccg 1021 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt 1081 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca 1141 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt 1201 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 1261 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg 1321 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 1381 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg 1441 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca 1501 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 1561 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag 1621 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca 1681 ctctcggcat ggacgagctg tacaagaagc ttacaagttt gtacaaaaaa gcaggctcca 1741 ccatgggaac caattcagtc gacatggagg acgagatgcc caagactcta tacgtcggta 1801 acctttccag agatgtgaca gaagctctaa ttctgcaact ctttagccag attggacctt 1861 gtaaaaactg caaaatgatt atggatacag ctggaaatga tccctattgt tttgtggagt 1921 ttcatgagca tcgtcatgca gctgcagcat tagctgctat gaatggacgg aagataatgg 1981 gtaaggaagt caaagtgaat tgggcaacaa cccctagcag tcaaaagaaa gatacaagca 2041 atcatttcca tgtctttgtt ggtgatctca gcccagaaat tacaactgaa gatataaaag 2101 ctgcttttgc accatttgga agaatatcag atgcccgagt ggtaaaagac atggcaacag 2161 gaaagtctaa gggatatggc tttgtctcct ttttcaacaa atgggatgct gaaaacgcca 2221 ttcaacagat gggtggccag tggcttggtg gaagacaaat cagaactaac tgggcaaccc 2281 gaaagcctcc cgctccaaag agtacatatg agtcaaatac caaacagcta tcatatgatg 2341 aggttgtaaa tcagtctagt ccaagcaact gtactgtata ctgtggaggt gttacttctg 2401 ggctaacaga acaactaatg cgtcagactt tttcaccatt tggacaaata atggaaattc 2461 gagtctttcc agataaagga tattcatttg ttcggttcaa ttcccatgaa agtgcagcac 2521 atgcaattgt ttctgttaat ggtactacca ttgaaggtca tgttgtgaaa tgctattggg 2581 gcaaagaaac tcttgatatg ataaatcccg tgcaacagca gaatcaaatt ggatatcccc 2641 aaccttatgg ccagtggggc cagtggtatg gaaatgcaca acaaattggc cagtatatgc 2701 ctaatggttg gcaagttcct gcatatggaa tgtatggcca ggcatggaac cagcaaggat 2761 ttaatcagac acagtcttct gcaccatgga tgggaccaaa ttatggagtg caaccgcctc 2821 aagggcaaaa tggcagcatg ttgcccaatc agccttctgg gtatcgagtg gcagggtatg 2881 aaacccagtg agcggccgca ctcgagatat ctagacccag ctttcttgta caaagtggtt 2941 gatatccagc acagtggcgg ccgctcaagt ctagagggcc cgcggttcga aggtaagcct 3001 atccctaacc ctctcctcgg tctcgattct acgctcgagt ctagagggcc cgtttaaacc 3061 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 3121 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 3181 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 3241 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 3301 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 3361 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 3421 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 3481 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 3541 ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga 3601 agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg 3661 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc 3721 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg 3781 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca 3841 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc 3901 tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg 3961 cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc 4021 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt 4081 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac 4141 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc 4201 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat 4261 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag 4321 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac 4381 ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc 4441 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg 4501 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa 4561 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt 4621 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc 4681 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc 4741 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt 4801 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 4861 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 4921 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 4981 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 5041 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 5101 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 5161 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 5221 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 5281 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 5341 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 5401 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 5461 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 5521 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 5581 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 5641 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 5701 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 5761 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 5821 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 5881 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 5941 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 6001 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 6061 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 6121 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 6181 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 6241 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 6301 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 6361 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 6421 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 6481 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 6541 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 6601 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 6661 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 6721 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 6781 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 6841 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 6901 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 6961 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 7021 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 7081 gttccgcgca catttccccg aaaagtgcca cctgacgtc //