LOCUS Exported 9487 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Tet-O-FUW-Nkx6.1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9487) AUTHORS Yang N, Zuchero JB, Ahlenius H, Marro S, Ng YH, Vierbuchen T, Hawkins JS, Geissler R, Barres BA, Wernig M TITLE Generation of oligodendroglial cells by direct lineage conversion. JOURNAL Nat Biotechnol. 2013 May;31(5):434-9. doi: 10.1038/nbt.2564. Epub 2013 Apr 14. PUBMED 23584610 REFERENCE 2 (bases 1 to 9487) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9487 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(320..331) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" misc_feature 1134..1722 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1187..1207) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1605..1616) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(1725..1741) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1726..1742) /label=pBluescriptKS /note="For pBluescript vector" LTR 2247..2427 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(2453..2470) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2459..2683 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2729..3157 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2816..2835) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3026..3047 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3166..3186) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3171..3500 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3351..3486 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3413..3432 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 3548..3595 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 3614..3988 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 4118..4239 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4155..4174) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4209..4228 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4288..4304) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4288..4304) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4301..4323) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4312..4328 /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(4336..4366) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4381..4402 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4519..4536) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4690..5278) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4770..4789) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5449..6309) /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 6072..6091 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6310..6414) /gene="bla" /label=AmpR promoter primer_bind complement(6489..6508) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 6680..7059 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 7061..7259 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 7214..7234 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 7277..7457 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 7504..7629 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8122..8355 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8540..8584 /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 8882..8999 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 9053..9323 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 9356..9393 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 9390..9414 /label=LNCX /note="Human CMV promoter, forward primer" ORIGIN 1 aattcgccac catgttagct gtgggggcga tggagggccc tcggcagagc gcgttcctgc 61 tcagcagccc gcccctggcc gccctgcaca gtatggccga gatgaagacc ccgctctacc 121 ccgccgctta tcccccgctg cccaccgggc ccccctcctc ctcgtcctcg tcctcctcgt 181 cctcgtcgcc ctccccacct ttgggctcac ataacccggg cggcttgaag cccccggccg 241 cggggggcct ctcgtccctg ggcagtcccc cgcagcagct ttcggcggcc accccacacg 301 gcatcaacga catcctgagc cggccctcta tgccggtggc ctcgggggcc gccctgccct 361 ccgcctcgcc ctccgggtct tcctcctcct cctcctcgtc cgcctccgcc acctcggcct 421 ctgcggcggc cgccgccgcc gctgctgctg ccgccgctgc cgcctcgtcg cccgctgggc 481 tgctggccgg cctgccccgc ttcagcagcc tgagccctcc gccaccgccg cccgggctct 541 actttagccc cagcgccgcg gctgtggccg ccgtgggccg gtaccccaag cccctggccg 601 agctgcccgg tcggacgccc atcttctggc ccggagtgat gcagagtccg ccgtggaggg 661 acgcgcgcct tgcctgtacc ccccatcaag gatccatttt gttggacaaa gatgggaaga 721 gaaaacacac cagacccacg ttctctggac agcaaatctt cgccctggag aagactttcg 781 aacaaacgaa gtacttggca ggaccagaga gagcacgctt ggcctattct ctggggatga 841 cggagagtca ggtcaaggtc tggttccaga accgcaggac caagtggaga aagaagcacg 901 cagccgagat ggccacggcc aagaagaagc aggactcgga gaccgagagg ctcaagggga 961 cttcggagaa tgaggaggat gacgacgatt acaacaaacc tctggacccg aactctgacg 1021 acgagaaaat cactcagctg ctgaaaaagc acaaatcgag cggtggcagc ctcctgctgc 1081 acgcgtcgga ggccgagggc tcgtcctgag aattcgatat caagcttatc gataatcaac 1141 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 1201 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 1261 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 1321 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 1381 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 1441 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 1501 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg 1561 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 1621 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc 1681 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc 1741 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt 1801 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac 1861 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 1921 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct 1981 accacacaca aggctacttc cctgattggc agaactacac accagggcca gggatcagat 2041 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag aaggtagaag 2101 aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat gggatggatg 2161 acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt catcacatgg 2221 cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg 2281 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc 2341 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct 2401 tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc agcctcgact 2461 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 2521 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 2581 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 2641 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga 2701 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg 2761 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct 2821 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 2881 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 2941 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 3001 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 3061 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta 3121 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt 3181 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3241 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3301 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 3361 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3421 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg 3481 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc 3541 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt 3601 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt 3661 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga 3721 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca 3781 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc 3841 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga 3901 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt 3961 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc 4021 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 4081 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa 4141 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 4201 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac 4261 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 4321 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 4381 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 4441 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 4501 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 4561 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 4621 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 4681 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 4741 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 4801 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 4861 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 4921 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 4981 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 5041 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 5101 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 5161 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 5221 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 5281 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 5341 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 5401 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 5461 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 5521 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 5581 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 5641 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 5701 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 5761 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 5821 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 5881 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 5941 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 6001 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 6061 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 6121 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 6181 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 6241 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 6301 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 6361 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 6421 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc 6481 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt 6541 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg 6601 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta 6661 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg 6721 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc 6781 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc 6841 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact 6901 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat 6961 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact 7021 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac 7081 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac 7141 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac 7201 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc 7261 gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 7321 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 7381 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 7441 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag 7501 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg 7561 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc 7621 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg 7681 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa 7741 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga 7801 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta 7861 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac 7921 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt 7981 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt 8041 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag 8101 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag 8161 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat 8221 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact 8281 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa 8341 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt 8401 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg 8461 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga 8521 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag 8581 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat 8641 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt 8701 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga 8761 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt 8821 agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca 8881 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa 8941 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc 9001 gggtttatta cagggacagc agagatccag tttggttact cgagtttacc actccctatc 9061 agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga 9121 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc 9181 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa 9241 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta 9301 ccactcccta tcagtgatag agaaaagtga aagtcgagct cggtacccgg gtcgagtagg 9361 cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg 9421 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc 9481 ggccccg //