LOCUS Exported 9206 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION doxycycline-inducible expression of mouse Pu.1 in mammalian cells. ACCESSION . VERSION . KEYWORDS pFUW-tetO-Pu.1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9206) AUTHORS Rosa FF, Pires CF, Kurochkin I, Ferreira AG, Gomes AM, Palma LG, Shaiv K, Solanas L, Azenha C, Papatsenko D, Schulz O, Reis e Sousa C, Pereira CF TITLE Direct reprogramming of fibroblasts into antigen-presenting dendritic cells. JOURNAL Sci Immunol. 2018 Dec 7;3(30). pii: 3/30/eaau4292. doi: 10.1126/sciimmunol.aau4292. PUBMED 30530727 REFERENCE 2 (bases 1 to 9206) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9206 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 167..284 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 341..611 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 644..682 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 679..703 /label=LNCX /note="Human CMV promoter, forward primer" misc_feature 1625..2213 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1678..1698) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2096..2107) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(2216..2232) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2217..2233) /label=pBluescriptKS /note="For pBluescript vector" LTR 2738..2918 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(2944..2961) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2950..3174 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3220..3648 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3307..3326) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3517..3538 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3657..3677) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3662..3991 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3842..3977 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3904..3923 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 4039..4086 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 4105..4479 /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 4609..4730 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4646..4665) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4700..4719 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4779..4795) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4779..4795) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4792..4814) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4803..4819 /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(4827..4857) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4872..4893 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5010..5027) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5181..5769) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5261..5280) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5940..6800) /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 6563..6582 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6801..6905) /gene="bla" /label=AmpR promoter primer_bind complement(6980..6999) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 7171..7550 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 7552..7750 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 7705..7725 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 7768..7948 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 7995..8120 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8613..8846 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 9031..9075 /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" ORIGIN 1 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat 61 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc 121 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag 181 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 241 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 301 acagcagaga tccagtttgg ttagatctcg agtttaccac tccctatcag tgatagagaa 361 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta 421 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat 481 agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg 541 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc 601 agtgatagag aaaagtgaaa gtcgagctcg gtacccgggt cgaggtaggc gtgtacggtg 661 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc 721 acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgcg gccccgaatt 781 catgttacag gcgtgcaaaa tggaagggtt ttccctcacc gcccctccat cggatgactt 841 ggttacttac gattcagagc tataccaacg tccaatgcat gactactact ccttcgtggg 901 cagcgatgga gaaagccata gcgatcacta ctgggatttc tccgcacacc atgtccacaa 961 caacgagttt gagaacttcc ctgagaacca cttcacagag ctgcagagtg tgcagccccc 1021 gcagctacag cagctctatc gccacatgga gctggaacag atgcacgtcc tcgatactcc 1081 catggtgcca ccccacaccg gcctcagtca ccaggtttcc tacatgcccc ggatgtgctt 1141 cccttatcaa accttgtccc cagcccacca gcagagctca gatgaggagg agggtgagag 1201 gcagagccct cccctggagg tgtctgatgg agaagctgat ggcttggagc ctgggccagg 1261 tcttctgcac ggggagacag gcagcaagaa aaagattcgc ctgtaccagt tcctgctgga 1321 cctgctgcgc agcggcgaca tgaaggacag catctggtgg gtggacaagg acaaaggtac 1381 cttccagttc tcgtccaagc acaaggaggc gctggcgcac cgctggggca tccagaaggg 1441 caaccgcaag aagatgacct accagaagat ggcgcgcgcg ctgcgcaact acggcaagac 1501 aggcgaggtg aagaaagtca agaagaagct cacctaccag ttcagcggcg aggtgctggg 1561 ccgtgggggc ctggccgagc ggcgcctccc gccccactga gaattcgata tcaagcttat 1621 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 1681 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 1741 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 1801 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 1861 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 1921 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 1981 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 2041 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 2101 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 2161 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata 2221 ccgtcgacct cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac 2281 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac 2341 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt 2401 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga 2461 tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc 2521 agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga 2581 gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca 2641 tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt 2701 tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat 2761 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 2821 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 2881 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat 2941 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt 3001 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat 3061 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg 3121 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg 3181 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat 3241 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 3301 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 3361 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 3421 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 3481 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 3541 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 3601 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa 3661 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 3721 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag 3781 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 3841 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 3901 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg 3961 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt 4021 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa 4081 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc 4141 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac 4201 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg 4261 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac 4321 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg 4381 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc 4441 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat 4501 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 4561 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt 4621 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 4681 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt 4741 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga 4801 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 4861 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 4921 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 4981 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 5041 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 5101 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 5161 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 5221 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 5281 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 5341 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 5401 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 5461 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 5521 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 5581 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 5641 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 5701 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 5761 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 5821 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 5881 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 5941 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 6001 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 6061 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 6121 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 6181 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 6241 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 6301 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 6361 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 6421 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 6481 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 6541 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 6601 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 6661 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 6721 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 6781 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 6841 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 6901 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc 6961 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc 7021 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa 7081 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc 7141 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt 7201 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta 7261 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga 7321 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt 7381 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta 7441 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg 7501 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt 7561 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc 7621 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat 7681 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct 7741 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg 7801 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 7861 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 7921 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag 7981 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag 8041 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga 8101 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa 8161 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca 8221 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac 8281 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat 8341 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg 8401 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg 8461 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat 8521 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga 8581 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga 8641 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta 8701 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat 8761 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa 8821 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc 8881 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat 8941 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc 9001 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat 9061 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta 9121 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata 9181 gtgaatagag ttaggcaggg atattc //