LOCUS Exported 9093 bp ds-DNA circular SYN 16-JUN-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Tet-O-FUW-Ascl1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9093) AUTHORS Vierbuchen T, Ostermeier A, Pang ZP, Kokubu Y, Sudhof TC, Wernig M TITLE Direct conversion of fibroblasts to functional neurons by defined factors. JOURNAL Nature. 2010 Feb 25. 463(7284):1035-41. PUBMED 20107439 REFERENCE 2 (bases 1 to 9093) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 16, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9093 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 298..415 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" CDS complement(713..757) /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 942..1175 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 1668..1793 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" LTR 1840..2020 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" promoter complement(2038..2236) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(2063..2083) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 2238..2617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" primer_bind 2789..2808 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 2883..2987 /gene="bla" /label=AmpR promoter CDS 2988..3848 /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(3206..3225) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4019..4607 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4508..4527 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4761..4778 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4895..4916 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4931..4961 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4969..4985 /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 4974..4996 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4993..5009 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4993..5009 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" polyA_signal 5058..5179 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5069..5088) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 5123..5142 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(5309..5683) /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" promoter complement(5702..5749) /label=EM7 promoter /note="synthetic bacterial promoter " promoter complement(5797..6126) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5811..5946 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(5865..5884) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 6111..6131 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin complement(6140..6568) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6250..6271) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6462..6481 /label=F1ori-R /note="F1 origin, reverse primer" polyA_signal 6614..6838 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 6827..6844 /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" LTR 6870..7050 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind 7555..7571 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 7556..7572 /label=KS primer /note="common sequencing primer, one of multiple similar variants" misc_feature 7575..8163 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS 7681..7692 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind 8090..8110 /label=WPRE-R /note="WPRE, reverse primer" regulatory 8880..8889 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(8972..8996) /label=LNCX /note="Human CMV promoter, forward primer" promoter complement(8993..9031) /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 9064..241 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" ORIGIN 1 gactttcact tttctctatc actgataggg agtggtaaac tcgactttca cttttctcta 61 tcactgatag ggagtggtaa actcgacttt cacttttctc tatcactgat agggagtggt 121 aaactcgact ttcacttttc tctatcactg atagggagtg gtaaactcga ctttcacttt 181 tctctatcac tgatagggag tggtaaactc gactttcact tttctctatc actgataggg 241 agtggtaaac tcgagatcta accaaactgg atctctgctg tccctgtaat aaacccgaaa 301 attttgaatt tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat 361 gtctactatt ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaattgtg 421 gatgaatact gccatttgtc tgcagaattg gcgcacgcag tgccgatccg ttcactaatc 481 gaatggatct gtctctgtct ctctctccac cttcttcttc tattccttcg ggcctgtcgg 541 gtcccctcgg ggttgggagg tgggtctgaa acgataatgg tgaatatccc tgcctaactc 601 tattcactat agaaagtaca gcaaaaacta ttcttaaacc taccaagcct cctactatca 661 ttatgaataa ttttatatac cacagccaat ttgttatgtt aaaccaattc cacaaacttg 721 cccatttatc taattccaat aattcttgtt cattcttttc ttgctggttt tgcgattctt 781 caattaagga gtgtattaag cttgtgtaat tgttaatttc tctgtcccac tccatccagg 841 tcgtgtgatt ccaaatctgt tccagagatt tattactcca actagcattc caaggcacag 901 cagtggtgca aatgagtttt ccagagcaac cccaaatccc caggagctgt tgatccttta 961 ggtatctttc cacagccagg attcttgcct ggagctgctt gatgccccag actgtgagtt 1021 gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc tgcactatac 1081 cagacaataa ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc 1141 ctgctgctcc caagaaccca aggaacaaag ctcctattcc cactgctctt ttttctctct 1201 gcaccactct tctctttgcc ttggtgggtg ctactcctaa tggttcaatt tttactactt 1261 tatatttata taattcactt ctccaattgt ccctcatatc tcctcctcca ggtctgaaga 1321 tcagcggccg cttgctgtgc ggtggtctta cttttgtttt gctcttcctc tatcttgtct 1381 aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact 1441 gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc 1501 agtatttgtc tacagccttc tgatgtttct aacaggccag gattaactgc gaatcgttct 1561 agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt 1621 aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac 1681 ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc 1741 ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct 1801 ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg 1861 actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta 1921 cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag 1981 agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca aaacgcgctg 2041 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg 2101 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca 2161 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac 2221 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta 2281 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg 2341 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt 2401 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga 2461 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca 2521 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta 2581 attgattact attaataact agtcaataat caatgtcaac gcgtatatct ggcccgtaca 2641 tcgcgaagca gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag 2701 ccttgccttg ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa 2761 gcagggagca gatactggct taactatgcg gcatcagagc agattgtact gagagtgcac 2821 cataggggat cgggagatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg 2881 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 2941 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac 3001 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 3061 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca 3121 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc 3181 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 3241 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac 3301 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 3361 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg 3421 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 3481 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 3541 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat 3601 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 3661 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 3721 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 3781 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 3841 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt 3901 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 3961 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 4021 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 4081 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 4141 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 4201 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 4261 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 4321 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 4381 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 4441 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 4501 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 4561 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 4621 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 4681 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc 4741 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac 4801 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc 4861 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg 4921 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat 4981 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg 5041 acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca 5101 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat 5161 aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc ccgcgctgga 5221 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg 5281 cggtggaatc gaaatctcgt agcacgtgtc agtcctgctc ctcggccacg aagtgcacgc 5341 agttgccggc cgggtcgcgc agggcgaact cccgccccca cggctgctcg ccgatctcgg 5401 tcatggccgg cccggaggcg tcccggaagt tcgtggacac gacctccgac cactcggcgt 5461 acagctcgtc caggccgcgc acccacaccc aggccagggt gttgtccggc accacctggt 5521 cctggaccgc gctgatgaac agggtcacgt cgtcccggac cacaccggcg aagtcgtcct 5581 ccacgaagtc ccgggagaac ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt 5641 cgcgcgcggt gagcaccgga acggcactgg tcaacttggc catggtttag ttcctcacct 5701 tgtcgtatta tactatgccg atatactatg ccgatgatta attgtcaaca cgtgctgatc 5761 agatccgaaa atggatatac aagctcccgg gagctttttg caaaagccta ggcctccaaa 5821 aaagcctcct cactacttct ggaatagctc agaggcagag gcggcctcgg cctctgcata 5881 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg 5941 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg 6001 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag 6061 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg 6121 acacacattc cacagaatta attcgcgtta aatttttgtt aaatcagctc attttttaac 6181 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg 6241 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa 6301 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt 6361 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt 6421 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga 6481 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc 6541 gcgcttaatg cgccgctaca gggcgcgtgg ggataccccc tagagcccca gctggttctt 6601 tccgcctcag aagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa 6661 tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga 6721 caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc accttccagg 6781 gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc 6841 gaggctgatc agcgggttta aacgggccct gctagagatt ttccacactg actaaaaggg 6901 tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac 6961 tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag agagctccca 7021 ggctcagatc tggtctaacc agagagaccc agtacagtcc ggatgcagct ctcgggccat 7081 gtgatgaaat gctaggcggc tgtcaaacct ccactctaat acttctctct ccgggtcatc 7141 catcccatgc aggctcacag ggtgtaacaa gcgggtgttc tctccttcat tggcttcttc 7201 taccttctct tgctcaactg gtactagctt gtagcaccat ccaaaggtca gtggatatct 7261 gatccctggc cctggtgtgt agttctgcca atcagggaag tagccttgtg tgtggtagat 7321 ccacagatca aggatatctt gtcttcgttg ggagtgaatt agcccttcca gtcccccctt 7381 ttcttttaaa aagtggctaa gatctacagc tgccttgtaa gtcattggtc ttaaaggtac 7441 ctgaggtgtg actggaaaac ccacctcctc ctcctcttgt gcttctagcc aggcacaatc 7501 agcattggta gctgctgtat tgctacttgt gattgctcca tgtttttcta ggtctcgagg 7561 tcgacggtat cgatgcgggg aggcggccca aagggagatc cgactcgtct gagggcgaag 7621 gcgaagacgc ggaagaggcc gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg 7681 attgagggcc gaagggacgt agcagaagga cgtcccgcgc agaatccagg tggcaacaca 7741 ggcgagcagc caaggaaagg acgatgattt ccccgacaac accacggaat tgtcagtgcc 7801 caacagccga gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc 7861 aatagggagg gggaaagcga aagtcccgga aaggagctga caggtggtgg caatgcccca 7921 accagtgggg gttgcgtcag caaacacagt gcacaccacg ccacgttgcc tgacaacggg 7981 ccacaactcc tcataaagag acagcaacca ggatttatac aaggaggaga aaatgaaagc 8041 catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa 8101 aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg 8161 attatcgata agcttgatat cgaattctca gaaccagttg gtaaagtcca gcagctcttg 8221 ttcctctggg ctaagagggt cgtaggatcc ctcgtcggag gagtaggacg agaccggaga 8281 acccgccata gagttcaagt cgttggagta gttgggggag atggtgggcg acaggacgcc 8341 cgcctgaaag gcagcgctca ccgcgtcgtg ctcgtccagc agctgctgca gcgcgcggat 8401 gtactcgacc gccgagcgca gcgtctccac cttgctcatc ttcttgttgg ccgcgccgtt 8461 ggggacatgc tcccggaggg tggcaaagcc caggttgacc aacttgaccc ggttgcgctc 8521 gcgctcgttg cggcgcgcca cggcggccgg ctgctgctgt ggcaggctgt agccgaagcc 8581 gctgaagttg agccggcgtt tgcagcgcat cagttccgga gaggacgagc gctggcgctt 8641 gacctgcttg gccgctgact tgtgaccgcc ccctgagggc tggctgtcgg ccaccgggct 8701 cagctgcggc gcctgctgcg gcggcgcctg cggctgttgc tgctgcgcgc tctgagctgc 8761 cgcggccgcc gccgctgccg ccgccgccgc ggtcgcaaag aagcaggctg cgggaggcag 8821 gaagggctgc gggggctgcg gctgctggcc ggctccactc tccatcttgc cagagctctc 8881 catggtggct agcgaattcg gggccgcgga ggctggatcg gtcccggtgt cttctatgga 8941 ggtcaaaaca gcgtggatgg cgtctccagg cgatctgacg gttcactaaa cgagctctgc 9001 ttatataggc ctcccaccgt acacgcctac ctcgacccgg gtaccgagct cgactttcac 9061 ttttctctat cactgatagg gagtggtaaa ctc //