LOCUS Exported 9237 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Retroviral expression of WT-STAT3. Please note that this plasmid contains WT-STAT3 tagged with FLAG, not GFP.. ACCESSION . VERSION . KEYWORDS pLEGFP-WT-STAT3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9237) AUTHORS Dasgupta M, Unal H, Willard B, Yang J, Karnik SS, Stark GR TITLE Critical role for lysine 685 in gene expression mediated by transcription factor unphosphorylated STAT3. JOURNAL J Biol Chem. 2014 Oct 31;289(44):30763-71. doi: 10.1074/jbc.M114.603894. Epub 2014 Sep 12. PUBMED 25217633 REFERENCE 2 (bases 1 to 9237) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9237 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 2882..2905 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" regulatory 2960..2969 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2966..3685 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2966..3682 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3011..3032) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3272..3291) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3619..3640 /label=EGFP-C /note="EGFP, forward primer" LTR 3763..4356 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(4459..4481) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 4567..4707 /label=bom /note="basis of mobility region from pBR322" primer_bind 4622..4641 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(4722..4739) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4893..5481) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4973..4992) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5652..6512) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6275..6294 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6513..6617) /gene="bla" /label=AmpR promoter primer_bind 6685..6703 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" LTR 7275..7450 /label=5' LTR (truncated) /note="truncated long terminal repeat from Moloney murine sarcoma virus" misc_feature 7513..7712 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 7913..8329 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 8259..8281 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 8297..8313 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 8373..9167 /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(8427..8446) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 9037..9056 /label=Neo-F /note="Neomycin resistance gene, forward primer" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 541 ccggactcag atctcgagct caagcttcag gatggcccaa tggaatcagc tacagcagct 601 tgacacacgg tacctggagc agctccatca gctctacagt gacagcttcc caatggagct 661 gcggcagttt ctggcccctt ggattgagag tcaagattgg gcatatgcgg ccagcaaaga 721 atcacatgcc actttggtgt ttcataatct cctgggagag attgaccagc agtatagccg 781 cttcctgcaa gagtcgaatg ttctctatca gcacaatcta cgaagaatca agcagtttct 841 tcagagcagg tatcttgaga agccaatgga gattgcccgg attgtggccc ggtgcctgtg 901 ggaagaatca cgccttctac agactgcagc cactgcggcc cagcaagggg gccaggccaa 961 ccaccccaca gcagccgtgg tgacggagaa gcagcagatg ctggagcagc accttcagga 1021 tgtccggaag agagtgcagg atctagaaca gaaaatgaaa gtggtagaga atctccagga 1081 tgactttgat ttcaactata aaaccctcaa gagtcaagga gacatgcaag atctgaatgg 1141 aaacaaccag tcagtgacca ggcagaagat gcagcagctg gaacagatgc tcactgcgct 1201 ggaccagatg cggagaagca tcgtgagtga gctggcgggg cttttgtcag cgatggagta 1261 cgtgcagaaa actctcacgg acgaggagct ggctgactgg aagaggcggc aacagattgc 1321 ctgcattgga ggcccgccca acatctgcct agatcggcta gaaaactgga taacgtcatt 1381 agcagaatct caacttcaga cccgtcaaca aattaagaaa ctggaggagt tgcagcaaaa 1441 agtttcctac aaaggggacc ccattgtaca gcaccggccg atgctggagg agagaatcgt 1501 ggagctgttt agaaacttaa tgaaaagtgc ctttgtggtg gagcggcagc cctgcatgcc 1561 catgcatcct gaccggcccc tcgtcatcaa gaccggcgtc cagttcacta ctaaagtcag 1621 gttgctggtc aaattccctg agttgaatta tcagcttaaa attaaagtgt gcattgacaa 1681 agactctggg gacgttgcag ctctcagagg atcccggaaa tttaacattc tgggcacaaa 1741 cacaaaagtg atgaacatgg aagaatccaa caacggcagc ctctctgcag aattcaaaca 1801 cttgaccctg agggagcaga gatgtgggaa tgggggccga gccaattgtg atgcttccct 1861 gattgtgact gaggagctgc acctgatcac ctttgagacc gaggtgtatc accaaggcct 1921 caagattgac ctagagaccc actccttgcc agttgtggtg atctccaaca tctgtcagat 1981 gccaaatgcc tgggcgtcca tcctgtggta caacatgctg accaacaatc ccaagaatgt 2041 aaactttttt accaagcccc caattggaac ctgggatcaa gtggccgagg tcctgagctg 2101 gcagttctcc tccaccacca agcgaggact gagcatcgag cagctgacta cactggcaga 2161 gaaactcttg ggacctggtg tgaattattc agggtgtcag atcacatggg ctaaattttg 2221 caaagaaaac atggctggca agggcttctc cttctgggtc tggctggaca atatcattga 2281 ccttgtgaaa aagtacatcc tggccctttg gaacgaaggg tacatcatgg gctttatcag 2341 taaggagcgg gagcgggcca tcttgagcac taagcctcca ggcaccttcc tgctaagatt 2401 cagtgaaagc agcaaagaag gaggcgtcac tttcacttgg gtggagaagg acatcagcgg 2461 taagacccag atccagtccg tggaaccata cacaaagcag cagctgaaca acatgtcatt 2521 tgctgaaatc atcatgggct ataagatcat ggatgctacc aatatcctgg tgtctccact 2581 ggtctatctc tatcctgaca ttcccaagga ggaggcattc ggaaagtatt gtcggccaga 2641 gagccaggag catcctgaag ctgacccagg tagcgctgcc ccatacctga agaccaagtt 2701 tatctgtgtg acaccaacga cctgcagcaa taccattgac ctgccgatgt ccccccgcac 2761 tttagattca ttgatgcagt ttggaaataa tggtgaaggt gctgaaccct cagcaggagg 2821 gcagtttgag tccctcacct ttgacatgga gttgacctcg gagtgcgcta cctcccccat 2881 ggactacaag gacgacgatg acaagtgaaa gcttcgaatt ctgcagtcga cggtaccgcg 2941 ggcccgggat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt 3001 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg 3061 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg 3121 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt 3181 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg 3241 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga 3301 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa 3361 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta 3421 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat 3481 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg 3541 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc 3601 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct 3661 cggcatggac gagctgtaca agtaaagcgg ccgcgactct agccttaatg gcctaacatc 3721 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg 3781 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact 3841 gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa 3901 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc 3961 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag 4021 aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt 4081 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt 4141 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc 4201 cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat 4261 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct 4321 gagtgattga ctacccgtca gcgggggtct ttcatttggg ggctcgtccg ggatcgggag 4381 acccctgccc agggaccacc gacccaccac cgggaggtaa gctggctgcc tcgcgcgttt 4441 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct 4501 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg 4561 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat 4621 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga 4681 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg 4741 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 4801 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 4861 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 4921 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 4981 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 5041 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 5101 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 5161 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 5221 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 5281 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta 5341 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 5401 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 5461 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 5521 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 5581 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 5641 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 5701 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 5761 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 5821 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 5881 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 5941 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt 6001 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 6061 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 6121 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 6181 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 6241 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact 6301 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 6361 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 6421 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 6481 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 6541 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 6601 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt 6661 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa 6721 ttcataccag atcaccgaaa actgtcctcc aaatgtgtcc ccctcacact cccaaattcg 6781 cgggcttctg cctcttagac cactctaccc tattccccac actcaccgga gccaaagccg 6841 cggcccttcc gtttctttgc ttttgaaaga ccccacccgt aggtggcaag ctagcttaag 6901 taacgccact ttgcaaggca tggaaaaata cataactgag aatagaaaag ttcagatcaa 6961 ggtcaggaac aaagaaacag ctgaatacca aacaggatat ctgtggtaag cggttcctgc 7021 cccggctcag ggccaagaac agatgagaca gctgagtgat gggccaaaca ggatatctgt 7081 ggtaagcagt tcctgccccg gctcggggcc aagaacagat ggtccccaga tgcggtccag 7141 ccctcagcag tttctagtga atcatcagat gtttccaggg tgccccaagg acctgaaaat 7201 gaccctgtac cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt 7261 ccgctctccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcttccga 7321 tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa 7381 tcgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cacgacgggg 7441 gtctttcatt tgggggctcg tccgggattt ggagacccct gcccagggac caccgaccca 7501 ccaccgggag gtaagctggc cagcaactta tctgtgtctg tccgattgtc tagtgtctat 7561 gtttgatgtt atgcgcctgc gtctgtacta gttagctaac tagctctgta tctggcggac 7621 ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac gtcccaggga 7681 ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga atccgacccc 7741 gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga 7801 atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat 7861 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac 7921 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca 7981 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc 8041 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt 8101 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt 8161 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa 8221 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac 8281 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg gaattccgat 8341 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 8401 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 8461 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 8521 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 8581 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 8641 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 8701 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 8761 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 8821 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 8881 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 8941 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 9001 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 9061 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 9121 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 9181 gttcgaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg //