LOCUS Exported 9512 bp ds-DNA circular SYN 27-MAY-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS FUW-TetO-Dmrt1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9512) AUTHORS Buganim Y, Itskovich E, Hu YC, Cheng AW, Ganz K, Sarkar S, Fu D, Welstead GG, Page DC, Jaenisch R TITLE Direct Reprogramming of Fibroblasts into Embryonic Sertoli-like Cells by Defined Factors. JOURNAL Cell Stem Cell. 2012 Sep 7;11(3):373-86. PUBMED 22958931 REFERENCE 2 (bases 1 to 9512) AUTHORS . TITLE Direct Submission JOURNAL Exported May 27, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9512 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 130..509 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 511..709 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 664..684 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 727..907 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 954..1079 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1572..1805 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1990..2034 /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 2332..2449 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 2506..2776 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 2809..2847 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2844..2868 /label=LNCX /note="Human CMV promoter, forward primer" misc_feature 4096..4684 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4149..4169) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4567..4578) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4687..4703) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4688..4704) /label=pBluescriptKS /note="For pBluescript vector" LTR 5209..5389 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(5415..5432) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5421..5645 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5691..6119 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5778..5797) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5988..6009 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6128..6148) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6133..6462 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6313..6448 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6375..6394 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 6510..6557 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 6576..6950 /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 7080..7201 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7117..7136) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7171..7190 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7250..7266) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7250..7266) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7263..7285) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7274..7290 /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(7298..7328) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7343..7364 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7481..7498) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7652..8240) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7732..7751) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8411..9271) /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 9034..9053 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9272..9376) /gene="bla" /label=AmpR promoter primer_bind complement(9451..9470) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt 61 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat 121 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag 181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct 241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc 301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg 361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat 421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc 721 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 781 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 841 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 901 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc 961 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga 1021 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca 1081 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa 1141 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag 1201 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac 1261 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct 1321 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg 1381 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga 1441 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt 1501 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga 1561 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc 1621 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag 1681 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg 1741 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag 1801 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat 1861 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg 1921 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac 1981 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat 2041 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc 2101 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga 2161 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa 2221 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga 2281 tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag 2341 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga 2401 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta 2461 cagggacagc agagatccag tttggttaga tctcgagttt accactccct atcagtgata 2521 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga 2581 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca 2641 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa 2701 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc 2761 ctatcagtga tagagaaaag tgaaagtcga gctcggtacc cgggtcgagg taggcgtgta 2821 cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc 2881 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccgcggcccc 2941 gaattgatgc cgaacgacga cacattcggc aagccctcta ccccgacgga ggttcctcac 3001 gctcccggag ccccgccgca gggcaaagcc ggaggttaca gcaaagctgc cggggcgatg 3061 gctggagcgg ctggaggctc gggcgcgggg ggcagcggag gtgcctcggg ctccgggccg 3121 tcgggcctgg gttctggaag caagaagtcc ccgaggctgc ccaaatgcgc tcgctgcagg 3181 aaccacggct acgcgtcgcc gctcaagggc cacaagcgct tctgcatgtg gcgggattgc 3241 cagtgcaaga agtgcagcct gattgcggag cgacagcggg tgatggccgc gcaggtggcc 3301 ctgagaagac agcaggccca ggaagaagaa ctgggtatca gccacccaat tccgctgccc 3361 agcgcagccg agctcctggt caaaagggag aataatgcca gcaacccgtg cctgatggcc 3421 gagaacagca gctctgccca gcccccaccg gccagcaccc caacccccgc tgcctcagag 3481 ggacgcatgg tcatccagga tattcctgct gtcaccagca gagggcatat ggagaacaca 3541 tctgacctgg tatcagaccc cgcctactac agcagctttt accagccttc tctgtttcct 3601 tactacaaca atctgtacaa ctacccccag tattccatgg ccttgtctgc tgagtcctcc 3661 tcaggggagg tggggaattc cctcggggga tcccccgtga agaacagcct gcggagtctc 3721 ccagcacctt acgtgcctgc tcagactgga aaccagtggc agatgaagac ctcagagagc 3781 cgccacccag tgagctccca gtaccggatg cactcatact acgggcctcc ctcctacctg 3841 ggccagagca tgtcccagat ctttaccttc gaggaaggcc cctcctactc agaagccaaa 3901 gccagtgtgt tctcgccccc cagcagtcaa gattctggct tggtctccct ctccagcagc 3961 tctccgatga gcaacgagag ctcgaaggga gttctggaat gtgagtctgc gtcctcggaa 4021 cctagcagct acgccgtcaa ccaggtcctg gaagaggatg aggacgagtg acaattcgat 4081 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt 4141 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 4201 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 4261 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 4321 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 4381 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 4441 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc 4501 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 4561 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 4621 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 4681 ccgcatcgat accgtcgacc tcgagaccta gaaaaacatg gagcaatcac aagtagcaat 4741 acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt 4801 tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt 4861 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa 4921 gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac 4981 acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca 5041 gttgagcaag agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct 5101 gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc 5161 cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg 5221 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct 5281 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt 5341 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta 5401 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc 5461 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga 5521 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca 5581 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc 5641 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg 5701 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 5761 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 5821 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg 5881 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 5941 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 6001 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 6061 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta 6121 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 6181 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 6241 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 6301 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 6361 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag 6421 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt 6481 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg 6541 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc 6601 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt 6661 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt 6721 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc 6781 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg 6841 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc 6901 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac 6961 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 7021 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 7081 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 7141 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 7201 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 7261 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 7321 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 7381 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 7441 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 7501 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 7561 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 7621 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 7681 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 7741 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 7801 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 7861 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 7921 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 7981 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 8041 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 8101 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 8161 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 8221 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 8281 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 8341 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 8401 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 8461 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 8521 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 8581 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 8641 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 8701 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 8761 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 8821 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 8881 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 8941 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 9001 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 9061 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 9121 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 9181 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 9241 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 9301 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 9361 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 9421 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag 9481 ccagtatctg ctccctgctt gtgtgttgga gg //