LOCUS Exported 8600 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses a chimeric SOX2-activator fusion. ACCESSION . VERSION . KEYWORDS pSN23 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8600) 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 8600) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8600 /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 2467..3055 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2520..2540) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2938..2949) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3208..3388 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" polyA_signal 3460..3581 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3497..3516) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3551..3570 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 3621..3756 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3683..3702 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3776..3795) /label=T7 /note="T7 promoter, forward primer" promoter complement(3777..3795) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3805..3822) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3805..3821) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3814..3836) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3963..4418 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4050..4069) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4260..4281 /label=F1ori-F /note="F1 origin, forward primer" promoter 4444..4548 /gene="bla" /label=AmpR promoter CDS 4549..5409 /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(4767..4786) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5580..6168 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6069..6088 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6322..6339 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6456..6477 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6492..6522 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6530..6546 /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 6535..6557 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6554..6570 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6554..6570 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 6589..6609 /label=T3 /note="T3 promoter, forward primer" promoter 6591..6609 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 6637..6863 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 6864..7044 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 7091..7216 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 7709..7942 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8127..8171 /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 8438..8555 /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 cgggatttac cccccacgac tccgccccct acggcgctct ggatatggcc 2401 gacttcgagt ttgagcagat gtttaccgat gcccttggaa ttgacgagta cggtgggtga 2461 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 2521 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 2581 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 2641 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 2701 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 2761 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 2821 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg 2881 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 2941 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 3001 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga 3061 attcgagctc ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact 3121 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc 3181 tgctagctcc ggatccggac ctgtactggg tctctctggt tagaccagat ctgagcctgg 3241 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 3301 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 3361 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat 3421 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat 3481 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 3541 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta 3601 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 3661 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc 3721 tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta 3781 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa 3841 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa 3901 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 3961 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 4021 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 4081 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 4141 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 4201 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 4261 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 4321 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 4381 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat 4441 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 4501 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa 4561 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 4621 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac 4681 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt 4741 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc 4801 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca 4861 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 4921 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag 4981 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 5041 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 5101 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa 5161 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 5221 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 5281 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 5341 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 5401 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat 5461 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 5521 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 5581 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 5641 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 5701 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc 5761 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 5821 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 5881 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 5941 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg 6001 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 6061 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 6121 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 6181 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 6241 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc 6301 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata 6361 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 6421 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag 6481 gcaccccagg ttttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga 6541 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct 6601 cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt 6661 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc 6721 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga 6781 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta 6841 agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc 6901 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc 6961 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt 7021 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa 7081 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga 7141 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga 7201 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg 7261 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga 7321 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat 7381 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa 7441 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc 7501 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc 7561 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat 7621 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg 7681 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag 7741 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt 7801 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt 7861 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat 7921 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca 7981 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca 8041 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa 8101 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat 8161 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca 8221 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga 8281 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg 8341 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca 8401 ttcgattagt gaacggatct cgacggtatc ggttaacttt taaaagaaaa ggggggattg 8461 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag 8521 aattacaaaa acaaattaca aaaattcaaa attttatcga tcacgagact agcctcgaga 8581 agcttgatat cgaattccac //