LOCUS       sequence-79842-e        5402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  conditional gene knockdowns in Trypanosoma brucei.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "EMBO J."; 
            date: "2007-11-28"; volume: "26(23)"; pages: "4856-66. Epub 2007 Nov
            1"
COMMENT     Sequence Label: pT7-stl
FEATURES             Location/Qualifiers
     source          1..5402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             join(3547..3615,3616..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
//
