LOCUS Exported 7838 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS TRMPV-ns SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7838) AUTHORS Zuber J, McJunkin K, Fellmann C, Dow LE, Taylor MJ, Hannon GJ, Lowe SW TITLE Toolkit for evaluating genes required for proliferation and survival using tetracycline-regulated RNAi. JOURNAL Nat Biotechnol. 2011 Jan . 29(1):79-83. PUBMED 21131983 REFERENCE 2 (bases 1 to 7838) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7838 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 310..513 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 464..484 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 514..689 /label=5' LTR (truncated) /note="truncated long terminal repeat from Moloney murine sarcoma virus" misc_feature 752..951 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1152..1568 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1498..1520 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1536..1552 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" protein_bind 1618..1888 /label=tetracycline response element /bound_moiety="tetracycline repressor TetR" /note="contains seven copies of the tetracycline operator tetO" promoter 1921..1959 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1956..1980 /label=LNCX /note="Human CMV promoter, forward primer" CDS 2058..2735 /codon_start=1 /product="improved tetrameric variant of DsRed fluorescent protein" /label=DsRed2 /note="mammalian codon-optimized" /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH LFL" primer_bind complement(2238..2258) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 2650..2673 /label=DsRed1-C /note="DsRed1, forward primer" ncRNA 2804..2925 /label=5' miR-30a /note="sequence upstream of the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" ncRNA 2953..2967 /label=miR-30a loop /note="loop from the the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" ncRNA 2995..3121 /label=3' miR-30a /note="sequence downstream of the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" promoter 3144..3643 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(3179..3202) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 3576..3595 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 3666..4385 /codon_start=1 /product="yellow fluorescent protein (YFP) with fast and efficient maturation (Nagai et al., 2002)" /label=Venus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3711..3732) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3972..3991) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4319..4340 /label=EGFP-C /note="EGFP, forward primer" LTR 4544..4969 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus" primer_bind complement(5072..5094) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5230..5250) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5235..5564 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5415..5550 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5477..5496 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 5586..5605 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(5686..5703) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5857..6445) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5937..5956) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6616..7476) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7239..7258 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7477..7581) /gene="bla" /label=AmpR promoter primer_bind 7649..7667 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" enhancer 7767..309 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc 61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac 121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata 181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgctctg gcattatgcc 241 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 301 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 361 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 421 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 481 cgtgtacggt gggaggtcta tataagcaga gctcaataaa agagcccaca acccctcact 541 cggcgcgcca gtcttccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct 601 cttgctgttt gcatccgaat cgtggtctcg ctgttccttg ggagggtctc ctctgagtga 661 ttgactaccc acgacggggg tctttcattt gggggctcgt ccgggatttg gagacccctg 721 cccagggacc accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt 781 ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg tctgtactag ttagctaact 841 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc 901 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt 961 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac 1021 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc 1081 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt 1141 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag 1201 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct 1261 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc 1321 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag 1381 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca 1441 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc 1501 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc 1561 taggcgccgg aattgaagat ctgggggatc gatcctctag agcatcgagt ttaccactcc 1621 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa 1681 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac 1741 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata 1801 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga 1861 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagctcggta cccgggtcga 1921 ggtaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc 1981 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc 2041 ctctcgacgg atccaccatg gcctcctccg agaacgtcat caccgagttc atgcgcttca 2101 aggtgcgcat ggagggcacc gtgaacggcc acgagttcga gatcgagggc gagggcgagg 2161 gccgccccta cgagggccac aacaccgtga agctgaaggt gaccaagggc ggccccctgc 2221 ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg tacgtgaagc 2281 accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc aagtgggagc 2341 gcgtgatgaa cttcgaggac ggcggcgtgg cgaccgtgac ccaggactcc tccctgcagg 2401 acggctgctt catctacaag gtgaagttca tcggcgtgaa cttcccctcc gacggccccg 2461 tgatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg 2521 gcgtgctgaa gggcgagacc cacaaggccc tgaagctgaa ggacggcggc cactacctgg 2581 tggagttcaa gtccatctac atggccaaga agcccgtgca gctgcccggc tactactacg 2641 tggacgccaa gctggacatc acctcccaca acgaggacta caccatcgtg gagcagtacg 2701 agcgcaccga gggccgccac cacctgttcc tgtagcggcc gcgactctag aatttactag 2761 tcatatgata agatctctcg actagggata acagggtaat tgtttgaatg aggcttcagt 2821 actttacaga atcgttgcct gcacatcttg gaaacacttg ctgggattac ttcttcaggt 2881 taacccaaca gaaggctcga gaaggtatat tgctgttgac agtgagcgca ggaattataa 2941 tgcttatcta tagtgaagcc acagatgtat agataagcat tataattcct atgcctactg 3001 cctcggaatt caaggggcta ctttaggagc aattatcttg tttactaaaa ctgaatacct 3061 tgctatctct ttgatacatt tttacaaagc tgaattaaaa tggtataaat taaatcactt 3121 ttttcaattc tagcaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc 3181 atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac 3241 acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc 3301 accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag 3361 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga 3421 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt 3481 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg 3541 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac 3601 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagctt 3661 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc 3721 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca 3781 cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc 3841 ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac cccgaccaca 3901 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca 3961 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca 4021 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg 4081 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga 4141 agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc 4201 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca 4261 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca 4321 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca 4381 agtaagaatt gttcaattgt aactacgtac gatgcatcga taccgtcgag atatcagtgg 4441 tccaggctct agttttgact caacaatatc accagctgaa gcctatagag tacgagccat 4501 agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct 4561 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac 4621 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagggtcg accctagaga 4681 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac 4741 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa 4801 agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac 4861 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg 4921 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt 4981 ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg taagctggct 5041 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg 5101 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg 5161 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtaga 5221 tccggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 5281 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 5341 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 5401 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 5461 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc 5521 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt actggcttaa 5581 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca 5641 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc 5701 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 5761 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 5821 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 5881 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 5941 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 6001 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 6061 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 6121 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 6181 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 6241 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac 6301 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 6361 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 6421 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 6481 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 6541 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 6601 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 6661 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 6721 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 6781 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 6841 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 6901 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt 6961 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 7021 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 7081 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 7141 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 7201 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag 7261 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 7321 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 7381 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 7441 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 7501 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 7561 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac 7621 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca 7681 agaattagct tggccattgc atacgttgta tccatatcat aatatgtaca tttatattgg 7741 ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc 7801 aattacgggg tcattagttc atagcccata tatggagt //