LOCUS Exported 9644 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral plasmids encoding human KLF4 with co-expressoin of GFP mediated by P2A.. ACCESSION . VERSION . KEYWORDS pLVH-EF1a-GFP-P2A-KLF4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9644) AUTHORS Shao Z, Zhang R, Khodadadi-Jamayran A, Chen B, Crowley MR, Festok MA, Crossman DK, Townes TM, Hu K TITLE The acetyllysine reader BRD3R promotes human nuclear reprogramming and regulates mitosis. JOURNAL Nat Commun. 2016 Mar 7;7:10869. doi: 10.1038/ncomms10869. PUBMED 26947130 REFERENCE 2 (bases 1 to 9644) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9644 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /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" misc_feature 2028..2143 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" promoter 2197..2408 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" intron 2409..3347 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3304..3324 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 3364..3373 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3370..4086 /codon_start=1 /product="Aequorea coerulescens GFP" /label=AcGFP1 /note="mammalian codon-optimized" /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF GFVTAAAITHGMDELYK" CDS 4114..4170 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" misc_feature 5612..6200 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5665..5685) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6083..6094) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6407..7040 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(7169..7185) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7169..7185) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7182..7204) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7193..7209 /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(7217..7247) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7262..7283 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7400..7417) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7571..8159) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7651..7670) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8330..9190) /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 8953..8972 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9191..9295) /gene="bla" /label=AmpR promoter polyA_signal 9343..9477 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(9380..9399) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 9434..9453 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgatgtgccc gtcagtgggc agagcgcaca tcgcccacag 2221 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg 2281 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag 2341 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca 2401 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc 2461 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc 2521 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg 2581 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc 2641 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg 2701 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta 2761 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg 2821 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc 2881 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc 2941 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag 3001 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa 3061 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc 3121 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg 3181 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag 3241 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca 3301 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgacccg 3361 gtcgccacca tggtgagcaa gggcgccgag ctgttcaccg gcatcgtgcc catcctgatc 3421 gagctgaatg gcgatgtgaa tggccacaag ttcagcgtga gcggcgaggg cgagggcgat 3481 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcctgtgccc 3541 tggcccaccc tggtgaccac cctgagctac ggcgtgcagt gcttctcacg ctaccccgat 3601 cacatgaagc agcacgactt cttcaagagc gccatgcctg agggctacat ccaggagcgc 3661 accatcttct tcgaggatga cggcaactac aagtcgcgcg ccgaggtgaa gttcgagggc 3721 gataccctgg tgaatcgcat cgagctgacc ggcaccgatt tcaaggagga tggcaacatc 3781 ctgggcaata agatggagta caactacaac gcccacaatg tgtacatcat gaccgacaag 3841 gccaagaatg gcatcaaggt gaacttcaag atccgccaca acatcgagga tggcagcgtg 3901 cagctggccg accactacca gcagaatacc cccatcggcg atggccctgt gctgctgccc 3961 gataaccact acctgtccac ccagagcgcc ctgtccaagg accccaacga gaagcgcgat 4021 cacatgatct acttcggctt cgtgaccgcc gccgccatca cccacggcat ggatgagctg 4081 tacaagtccg gactcagatc tcgaggatct ggagcaacaa acttctcact actcaaacaa 4141 gcaggtgacg tggaggagaa tcccggcccc atggctgtca gcgacgcgct gctcccatct 4201 ttctccacgt tcgcgtctgg cccggcggga agggagaaga cactgcgtca agcaggtgcc 4261 ccgaataacc gctggcggga ggagctctcc cacatgaagc gacttccccc agtgcttccc 4321 ggccgcccct atgacctggc ggcggcgacc gtggccacag acctggagag cggcggagcc 4381 ggtgcggctt gcggcggtag caacctggcg cccctacctc ggagagagac cgaggagttc 4441 aacgatctcc tggacctgga ctttattctc tccaattcgc tgacccatcc tccggagtca 4501 gtggccgcca ccgtgtcctc gtcagcgtca gcctcctctt cgtcgtcgcc gtcgagcagc 4561 ggccctgcca gcgcgccctc cacctgcagc ttcacctatc cgatccgggc cgggaacgac 4621 ccgggcgtgg cgccgggcgg cacgggcgga ggcctcctct atggcaggga gtccgctccc 4681 cctccgacgg ctcccttcaa cctggcggac atcaacgacg tgagcccctc gggcggcttc 4741 gtggccgagc tcctgcggcc agaattggac ccggtgtaca ttccgccgca gcagccgcag 4801 ccgccaggtg gcgggctgat gggcaagttc gtgctgaagg cgtcgctgag cgcccctggc 4861 agcgagtacg gcagcccgtc ggtcatcagc gtcagcaaag gcagccctga cggcagccac 4921 ccggtggtgg tggcgcccta caacggcggg ccgccgcgca cgtgccccaa gatcaagcag 4981 gaggcggtct cttcgtgcac ccacttgggc gctggacccc ctctcagcaa tggccaccgg 5041 ccggctgcac acgacttccc cctggggcgg cagctcccca gcaggactac cccgaccctg 5101 ggtcttgagg aagtgctgag cagcagggac tgtcaccctg ccctgccgct tcctcccggc 5161 ttccatcccc acccggggcc caattaccca tccttcctgc ccgatcagat gcagccgcaa 5221 gtcccgccgc tccattacca agagctcatg ccacccggtt cctgcatgcc agaggagccc 5281 aagccaaaga ggggaagacg atcgtggccc cggaaaagga ccgccaccca cacttgtgat 5341 tacgcgggct gcggcaaaac ctacacaaag agttcccatc tcaaggcaca cctgcgaacc 5401 cacacaggtg agaaacctta ccactgtgac tgggacggct gtggatggaa attcgcccgc 5461 tcagatgaac tgaccaggca ctaccgtaaa cacacggggc accgcccgtt ccagtgccaa 5521 aaatgcgacc gagcattttc caggtcggac cacctcgcct tacacatgaa gaggcatttt 5581 taaggatcca ccggatctag gcgtctggaa caatcaacct ctggattaca aaatttgtga 5641 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 5701 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa 5761 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 5821 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 5881 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 5941 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 6001 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg 6061 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 6121 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc 6181 cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga aaaacatgga 6241 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa 6301 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac 6361 aaggcagctg tagatcttag ccacttttta aaagaaaaga ggggactgga agggctaatt 6421 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc 6481 cctgattagc agaactacac accagggcca ggggtcagat atccactgac ctttggatgg 6541 tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac 6601 accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta 6661 gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag 6721 tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca 6781 gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca 6841 gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 6901 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 6961 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 7021 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 7081 tgcaaagaaa tgaatatcag agagtgagag gccttgacat tgctagcgtt ttaccgtcga 7141 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 7201 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 7261 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 7321 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 7381 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 7441 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 7501 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 7561 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 7621 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 7681 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 7741 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 7801 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 7861 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 7921 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 7981 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 8041 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 8101 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 8161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 8221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 8281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 8341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 8401 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 8461 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 8521 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 8581 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 8641 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 8701 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 8761 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 8821 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 8881 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 8941 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 9001 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 9061 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 9121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 9181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 9241 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 9301 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcaacttgtt tattgcagct 9361 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 9421 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac 9481 tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact 9541 gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt 9601 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt //