LOCUS Exported 9050 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION expresses a chimeric KLF4-activator fusion. ACCESSION . VERSION . KEYWORDS pSN31 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9050) AUTHORS Narayan S, Bryant G, Shah S, Berrozpe G, Ptashne M TITLE OCT4 and SOX2 Work as Transcriptional Activators in Reprogramming Human Fibroblasts. JOURNAL Cell Rep. 2017 Aug 15;20(7):1585-1596. doi: 10.1016/j.celrep.2017.07.071. PUBMED 28813671 REFERENCE 2 (bases 1 to 9050) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9050 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..511 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 430..448 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 526..1233 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(671..689) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 939..958 /label=mCherry-F /note="mCherry, forward primer" CDS 1243..1299 /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 2917..3505 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2970..2990) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3388..3399) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3658..3838 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" polyA_signal 3910..4031 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3947..3966) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4001..4020 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4071..4206 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4133..4152 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4226..4245) /label=T7 /note="T7 promoter, forward primer" promoter complement(4227..4245) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4255..4272) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4255..4271) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4264..4286) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4413..4868 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4500..4519) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4710..4731 /label=F1ori-F /note="F1 origin, forward primer" promoter 4894..4998 /gene="bla" /label=AmpR promoter CDS 4999..5859 /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(5217..5236) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6030..6618 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6519..6538 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6772..6789 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6906..6927 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6942..6972 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6980..6996 /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 6985..7007 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7004..7020 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7004..7020 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 7039..7059 /label=T3 /note="T3 promoter, forward primer" promoter 7041..7059 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 7087..7313 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 7314..7494 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 7541..7666 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8159..8392 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8577..8621 /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 8888..9005 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc 61 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc 121 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc 181 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac 241 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc 301 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag 361 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct 421 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct 481 cgttgaccga atcaccgacc tctctcccca gggggatcca ccggtatggt gagcaagggc 541 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc 601 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc 661 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc 721 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc 781 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag 841 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac 901 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc 961 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag 1021 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc 1081 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac 1141 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc 1201 cactccaccg gcggcatgga cgagctgtac aagggatctg gagcaacaaa cttctcacta 1261 ctcaaacaag caggtgacgt ggaggagaat cccggcccta tggctgtcag cgacgcgctg 1321 ctcccatctt tctccacgtt cgcgtctggc ccggcgggaa gggagaagac actgcgtcaa 1381 gcaggtgccc cgaataaccg ctggcgggag gagctctccc acatgaagcg acttccccca 1441 gtgcttcccg gccgccccta tgacctggcg gcggcgaccg tggccacaga cctggagagc 1501 ggcggagccg gtgcggcttg cggcggtagc aacctggcgc ccctacctcg gagagagacc 1561 gaggagttca acgatctcct ggacctggac tttattctct ccaattcgct gacccatcct 1621 ccggagtcag tggccgccac cgtgtcctcg tcagcgtcag cctcctcttc gtcgtcgccg 1681 tcgagcagcg gccctgccag cgcgccctcc acctgcagct tcacctatcc gatccgggcc 1741 gggaacgacc cgggcgtggc gccgggcggc acgggcggag gcctcctcta tggcagggag 1801 tccgctcccc ctccgacggc tcccttcaac ctggcggaca tcaacgacgt gagcccctcg 1861 ggcggcttcg tggccgagct cctgcggcca gaattggacc cggtgtacat tccgccgcag 1921 cagccgcagc cgccaggtgg cgggctgatg ggcaagttcg tgctgaaggc gtcgctgagc 1981 gcccctggca gcgagtacgg cagcccgtcg gtcatcagcg tcagcaaagg cagccctgac 2041 ggcagccacc cggtggtggt ggcgccctac aacggcgggc cgccgcgcac gtgccccaag 2101 atcaagcagg aggcggtctc ttcgtgcacc cacttgggcg ctggaccccc tctcagcaat 2161 ggccaccggc cggctgcaca cgacttcccc ctggggcggc agctccccag caggactacc 2221 ccgaccctgg gtcttgagga agtgctgagc agcagggact gtcaccctgc cctgccgctt 2281 cctcccggct tccatcccca cccggggccc aattacccat ccttcctgcc cgatcagatg 2341 cagccgcaag tcccgccgct ccattaccaa gagctcatgc cacccggttc ctgcatgcca 2401 gaggagccca agccaaagag gggaagacga tcgtggcccc ggaaaaggac cgccacccac 2461 acttgtgatt acgcgggctg cggcaaaacc tacacaaaga gttcccatct caaggcacac 2521 ctgcgaaccc acacaggtga gaaaccttac cactgtgact gggacggctg tggatggaaa 2581 ttcgcccgct cagatgaact gaccaggcac taccgtaaac acacggggca ccgcccgttc 2641 cagtgccaaa aatgcgaccg agcattttcc aggtcggacc acctcgcctt acacatgaag 2701 aggcattttc acttagacgg cgaggacgtg gcgatggcgc atgccgacgc gctagacgat 2761 ttcgatctgg acatgttggg ggacggggat tccccgggtc cgggatttac cccccacgac 2821 tccgccccct acggcgctct ggatatggcc gacttcgagt ttgagcagat gtttaccgat 2881 gcccttggaa ttgacgagta cggtgggtga gtcgacaatc aacctctgga ttacaaaatt 2941 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 3001 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 3061 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 3121 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 3181 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 3241 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 3301 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg 3361 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 3421 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 3481 atctcccttt gggccgcctc cccgcctgga attcgagctc ggtaccttta agaccaatga 3541 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc 3601 taattcactc ccaacgaaga caagatatcc tgctagctcc ggatccggac ctgtactggg 3661 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 3721 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 3781 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 3841 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 3901 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 3961 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 4021 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc 4081 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 4141 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 4201 gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc 4261 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc 4321 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc 4381 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc 4441 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 4501 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 4561 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 4621 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 4681 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 4741 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg 4801 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct 4861 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc 4921 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa 4981 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt 5041 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct 5101 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc 5161 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta 5221 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac 5281 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc 5341 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac 5401 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg 5461 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac 5521 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc 5581 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt 5641 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga 5701 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc 5761 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag 5821 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca 5881 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc 5941 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 6001 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 6061 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 6121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt 6181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 6241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 6301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 6361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 6421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 6481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 6541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 6601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 6661 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 6721 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 6781 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg 6841 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac 6901 gcaattaatg tgagttagct cactcattag gcaccccagg ttttacactt tatgcttccg 6961 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac 7021 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa 7081 gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc 7141 aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt 7201 acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg 7261 aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc 7321 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 7381 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 7441 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 7501 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg 7561 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat 7621 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg 7681 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa 7741 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct 7801 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac 7861 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca 7921 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa 7981 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga 8041 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag 8101 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag 8161 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga 8221 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc 8281 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc 8341 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt 8401 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta 8461 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta 8521 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga 8581 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa 8641 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa 8701 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat 8761 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag 8821 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc 8881 ggttaacttt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag 8941 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa 9001 attttatcga tcacgagact agcctcgaga agcttgatat cgaattccac //