LOCUS Exported 8495 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses a chimeric SOX2-activator fusion. ACCESSION . VERSION . KEYWORDS pSN24 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8495) 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 8495) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8495 /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..1242 /codon_start=1 /product="enhanced variant of YFP (Heikal et al., 2001)" /label=Citrine /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(571..592) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(832..851) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1179..1200 /label=EGFP-C /note="EGFP, forward primer" CDS 1252..1308 /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 2362..2950 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2415..2435) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2833..2844) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3103..3283 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" polyA_signal 3355..3476 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3392..3411) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3446..3465 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 3516..3651 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3578..3597 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3671..3690) /label=T7 /note="T7 promoter, forward primer" promoter complement(3672..3690) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3700..3717) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3700..3716) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3709..3731) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3858..4313 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3945..3964) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4155..4176 /label=F1ori-F /note="F1 origin, forward primer" promoter 4339..4443 /gene="bla" /label=AmpR promoter CDS 4444..5304 /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(4662..4681) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5475..6063 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5964..5983 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6217..6234 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6351..6372 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6387..6417 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6425..6441 /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 6430..6452 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6449..6465 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6449..6465 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 6484..6504 /label=T3 /note="T3 promoter, forward primer" promoter 6486..6504 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 6532..6758 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 6759..6939 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 6986..7111 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 7604..7837 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8022..8066 /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 8333..8450 /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 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc 601 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg 661 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc 721 ggctacggcc tgatgtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc 781 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc 841 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag 901 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac 961 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac 1021 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag 1081 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag 1141 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg 1201 accgccgccg ggatcactct cggcatggac gagctgtaca agggatctgg agcaacaaac 1261 ttctcactac tcaaacaagc aggtgacgtg gaggagaatc ccggccctat gtacaacatg 1321 atggagacgg agctgaagcc gccgggcccg cagcaaactt cggggggcgg cggcggcaac 1381 tccaccgcgg cggcggccgg cggcaaccag aaaaacagcc cggaccgcgt caagcggccc 1441 atgaatgcct tcatggtgtg gtcccgcggg cagcggcgca agatggccca ggagaacccc 1501 aagatgcaca actcggagat cagcaagcgc ctgggcgccg agtggaaact tttgtcggag 1561 acggagaagc ggccgttcat cgacgaggct aagcggctgc gagcgctgca catgaaggag 1621 cacccggatt ataaataccg gccccggcgg aaaaccaaga cgctcatgaa gaaggataag 1681 tacacgctgc ccggcgggct gctggccccc ggcggcaata gcatggcgag cggggtcggg 1741 gtgggcgccg gcctgggcgc gggcgtgaac cagcgcatgg acagttacgc gcacatgaac 1801 ggctggagca acggcagcta cagcatgatg caggaccagc tgggctaccc gcagcacccg 1861 ggcctcaatg cgcacggcgc agcgcagatg cagcccatgc accgctacga cgtgagcgcc 1921 ctgcagtaca actccatgac cagctcgcag acctacatga acggctcgcc cacctacagc 1981 atgtcctact cgcagcaggg cacccctggc atggctcttg gctccatggg ttcggtggtc 2041 aagtccgagg ccagctccag cccccctgtg gttacctctt cctcccactc cagggcgccc 2101 tgccaggccg gggacctccg ggacatgatc agcatgtatc tccccggcgc cgaggtgccg 2161 gaacccgccg cccccagcag acttcacatg tcccagcact accagagcgg cccggtgccc 2221 ggcacggcca ttaacggcac actgcccctc tcacacatgc acttagacgg cgaggacgtg 2281 gcgatggcgc atgccgacgc gctagacgat ttcgatctgg acatgttggg ggacggggat 2341 tccccgggtc cgtgagtcga caatcaacct ctggattaca aaatttgtga aagattgact 2401 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 2461 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 2521 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 2581 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 2641 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 2701 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg 2761 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 2821 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 2881 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 2941 gcctccccgc ctggaattcg agctcggtac ctttaagacc aatgacttac aaggcagctg 3001 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac 3061 gaagacaaga tatcctgcta gctccggatc cggacctgta ctgggtctct ctggttagac 3121 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa 3181 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag 3241 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat 3301 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg 3361 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 3421 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat 3481 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 3541 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 3601 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt 3661 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg 3721 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt 3781 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag 3841 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 3901 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 3961 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 4021 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 4081 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 4141 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 4201 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 4261 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc 4321 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 4381 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 4441 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt 4501 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt 4561 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc 4621 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta 4681 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac 4741 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa 4801 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg 4861 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc 4921 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg 4981 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta 5041 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg 5101 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg 5161 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc 5221 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt 5281 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt 5341 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc 5401 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 5461 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 5521 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 5581 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag 5641 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 5701 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 5761 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 5821 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 5881 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 5941 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 6001 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 6061 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 6121 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga 6181 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg 6241 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc 6301 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt 6361 tagctcactc attaggcacc ccaggtttta cactttatgc ttccggctcg tatgttgtgt 6421 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag 6481 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt 6541 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag 6601 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat 6661 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca 6721 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga 6781 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct 6841 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat 6901 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt 6961 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg 7021 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg 7081 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg 7141 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt 7201 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga 7261 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact 7321 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa 7381 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc 7441 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa 7501 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg 7561 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg 7621 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg 7681 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg 7741 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc 7801 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa 7861 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac 7921 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct 7981 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat 8041 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt 8101 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg 8161 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc 8221 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag 8281 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcggtta acttttaaaa 8341 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag 8401 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt atcgatcacg 8461 agactagcct cgagaagctt gatatcgaat tccac //