LOCUS Exported 5402 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION conditional gene knockdowns in Trypanosoma brucei. ACCESSION . VERSION . KEYWORDS pT7-stl SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5402) AUTHORS Brandenburg J, Schimanski B, Nogoceke E, Nguyen TN, Padovan JC, Chait BT, Cross GA, Gunzl A TITLE Multifunctional class I transcription in Trypanosoma brucei depends on a novel protein complex. JOURNAL EMBO J. 2007 Nov 28;26(23):4856-66. Epub 2007 Nov 1. PUBMED 17972917 REFERENCE 2 (bases 1 to 5402) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5402 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(1082..1456) /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(1600..1618) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 1629..1647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1652..1670 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 1671..1689 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind complement(1830..1848) /label=LucNrev /note="Firefly luciferase, reverse primer" terminator 1944..1991 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 2077..2124 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(2638..2656) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2639..2656) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2947..2966) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(3177..3196) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3296..3318 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3356..3374) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3442..3546 /gene="bla" /label=AmpR promoter CDS 3547..4407 /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(3765..3784) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4578..5166 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5067..5086 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5320..5337 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gaattcgagc tcatatagtt ggtatgtatt ctaattccag actactggcg tggataaaca 61 tgtcccctga ttaaaaggaa agattccata gcccattagt gcaaagataa ttggtacacc 121 gtcaaaaaca catgggagac cacacgatac gacactgtga ccgtagcatc aattccgcac 181 tcgatataac tctatcgaag aacttccatg gtacaaactg gtcgccacgg tctgctccac 241 acggagatca tcgtatcatt tttatcgata gcggccgcta tcgatgtatg ccttggccct 301 gatggcatgc caatttcact acaacggact atgtggaccc cgttatcatg gaaatgcgct 361 agttggagga agttagaccg cgccggaaaa gagaggggta gagaaaatga caacttggaa 421 gatatccaca cacgcacggt gaaacgttag caacaattat tagggaagca cgcttgccct 481 agtcccacga gtaaccaaga ctccaaaagc ctttctggca cagagagcga gccgaaatgg 541 aaaagagaaa caatgcctgc actaacacta ctgagcgatt cgcctcgccg cggaggaccg 601 aatactaata acgacacttg cggtcaaaaa gtagaagaac aaatgctcaa cgatgagtga 661 atcaggttag ggtagttgga aaattataca gaatgtcttt ggcaacacac cggcnnnctt 721 gcatgcctgc aaggccttgc agaatactgc atagataaca aacgcatcaa acacaactgg 781 tggcactatt tcaatcatgt cgacacacca agacgagaaa atttgagtta aacaagatgc 841 ctacaaccct atatttatta tcttcaagtg atcttctctt ctcttgtgcg ctgtacgtaa 901 atgtgttgca aataaggcat agggctgagt acaggcacca cacgcgagga aaatcataag 961 cactagccgt gtcaactcac agcgactaca gaacaatttt ggccacacaa cccggtgtta 1021 ggatctccga ggcctgggac ccgtgggccg ccgtcggacc ggcggtgttg gtcggcgtcg 1081 gtcagtcctg ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga 1141 actcccgccc ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga 1201 agttcgtgga cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca 1261 cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca 1321 cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc 1381 ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac 1441 tggtcaactt ggccatggtg atatagctta ttttatggca gcaacgagac cttacgtaaa 1501 gcaaaactaa ataagagcgg agactgcaat gcagagtaaa aaaaaaaaaa acatgaattt 1561 cagaagacct tgctgtgccc ccggtacggg ctagatctcc ctatagtgag tcgtattaat 1621 caggtaccta atacgactca ctatagggat ctccctatca gtgatagaga tccctatcag 1681 tgatagagaa gcttggaatt cctttgtgtt acattcttga atgtcgctcg cagtgacatt 1741 agcattccgg tactgttggt aaaatggaag acgccaaaaa cataaagaaa ggcccggcgc 1801 cattctatcc tctagaggat ggaaccgctg gagagcaact gcataaggct atgaagagat 1861 acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg aacatcacgt 1921 acgacgcgtc cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga 1981 ggggtttttt gctgaaagga ggaactatat ccggataggg ctgctaacaa agcccgaaag 2041 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct 2101 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggcgc gtggtacatc 2161 aatgggatca tatcttgctg cctcccgcag ctcaccgtgc gagctgccgg gaccccgttt 2221 ctcataggcg tgtgcacctt ccgtctgaga cctgtaaatg gttaaaaagg aatatataat 2281 taccttttga aaagtggtaa aaaacgaata tatctttttt tttggtttta atacgctctt 2341 ttgtgtgtat gagaggaata aataagtgtg tgtgtgtgtg tgtgttgttg ttgttgaaaa 2401 aatgaaggac acgaaagcgc tcattaaggg tggggataca ccaggcataa ttatccgaaa 2461 atattttgat gcaccaaata agtgaacata ctgacgaaat caaagagttg gaggatggat 2521 agggaggcct cattggcagt ggtaaattga tttgacttga atatgatacc ctcttactgt 2581 ttcattgcca ctacaagttg gtttccttcc cctgcaggca tgcaagctag cttgtattct 2641 atagtgtcac ctaaatcgta tgtgtatgat acataaggtt atgtattaat tgtagccgcg 2701 ttctaacgac aatatgtaca agcctaattg tgtagcatct ggcttactga agcagaccct 2761 atcatctctc tcgtaaactg ccgtcagagt cggtttggtt ggacgaacct tctgagtttc 2821 tggtaacgcc gttccgcacc ccggaaatgg tcagcgaacc aatcagcagg gtcatcgcta 2881 gccagatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg 2941 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca 3001 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg 3061 ccatctcctt gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg 3121 ctgcttccta atgcaggagt cgcataaggg agagcgtcga tatggtgcac tctcagtaca 3181 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg 3241 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg 3301 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc 3361 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt 3421 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 3481 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 3541 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 3601 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 3661 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 3721 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 3781 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 3841 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 3901 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 3961 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 4021 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 4081 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 4141 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 4201 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 4261 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 4321 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 4381 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 4441 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 4501 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 4561 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 4621 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 4681 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 4741 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 4801 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 4861 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 4921 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 4981 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 5041 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 5101 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 5161 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 5221 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 5281 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 5341 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc 5401 ag //