LOCUS       40924_44304        9177 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTrypRNAiGate, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9177)
  AUTHORS   Kalidas S, Li Q, Phillips MA.
  TITLE     A Gateway((R)) compatible vector for gene silencing in bloodstream 
            form Trypanosoma brucei
  JOURNAL   Mol. Biochem. Parasitol. 178 (1-2), 51-55 (2011)
  PUBMED    21420443
REFERENCE   2  (bases 1 to 9177)
  AUTHORS   Kalidas S, Li Q, Phillips M.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-MAR-2011) Department of Pharmacology, UT Southwestern 
            Medical Center, 6001 Forest Park Rd, Dallas, TX 75390-9041, USA
REFERENCE   3  (bases 1 to 9177)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9177)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. 
            Biochem. Parasitol. 178 (1-2), 51-55 (2011)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-MAR-2011) Department of Pharmacology, UT Southwestern Medical 
            Center, 6001 Forest Park Rd, Dallas, TX 75390-9041, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(100..732)
                     /label=rRNA locus intergenic fragment
                     /note="rRNA locus intergenic fragment"
     CDS             complement(1085..1456)
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(1600..1618)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1883..1901
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1904..1922
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2053..2177
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2202..2232
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2286..2942
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3287..3589
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3633..3757)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    4213..4337
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4381..4683)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(5028..5684)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(5738..5768)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    complement(5793..5917)
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(6413..6431)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        7217..7321
                     /label=AmpR promoter
     CDS             7322..8179
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHSMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8353..8941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 caggtaccgt cattggggtt aagcggaaag gtgtgtgtca gtaggttgtg aggtgaaagc
     1681 gttttcagat gcatagtgag cttaatgtcc ttttcacagt atatcgtgtc tgataggtat
     1741 ctcttattag tatagtcgaa tactagtcaa tagtgcgttt tgtgcaaaat gtccattttg
     1801 tggcagtgat ggggttgttt tatgctattc cgtgtctctg ggtgggcgtg cattgaaaat
     1861 aggggttatc gggtgaggga tctccctatc agtgatagag atctccctat cagtgataga
     1921 gatccctgag tactgagttt aacatgttct cgtcccgggc tgcacgcgcc ttcgagtttt
     1981 ttttcctttt ccccattttt ttcaacttga agacttcaat tacaccaaaa agtaaaattc
     2041 acaagcttat caacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
     2101 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     2161 catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg
     2221 gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag
     2281 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     2341 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     2401 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     2461 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     2521 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     2581 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     2641 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     2701 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     2761 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     2821 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     2881 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     2941 cgtaaagatc tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     3001 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta
     3061 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     3121 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     3181 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     3241 attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt
     3301 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     3361 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     3421 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     3481 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     3541 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     3601 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     3661 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     3721 tttacgtttc tcgttcagct ttcttgtaca aagtggttga tcgaggggca cacctagtcc
     3781 tcttcacccg ggtcgcaagt aagtccatga taaacaaaag ggagtagtac ttaaactcgg
     3841 cactacgaat actctaagcg gatgactacc tcgttctcag aagacgaaca tcggccgctg
     3901 actttccaag acctcctcct cagggtcctc cttcactttg accccctgaa ccccctggtc
     3961 cctgaggtcc tcctccagac ggggttgacc gagactttga agccgacgtt caggacgagg
     4021 accgagctca ggaatctcca gtagggggtg aagacgatct gcaccagtct ttacggacac
     4081 tagagaagta aggagacctg tttgatccgg agaccgcgac accgggaccc taacaccccg
     4141 aaacaccgga gcacaggagg taggacagat aagagtggga ttagataggg accgatgctg
     4201 agttcgggat caaccacttt gtacaagaaa gctgaacgag aaacgtaaaa tgatataaat
     4261 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     4321 catatccagt cactatggtc gacctgcaga ctggctgtgt ataagggagc ctgacattta
     4381 tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc
     4441 agccacttct tccccgataa cggagaccgg cacactggcc atatcggtgg tcatcatgcg
     4501 ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggagactt tatctgacag
     4561 cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact
     4621 ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg
     4681 catttcacca gcccctgttc tcgtcagcaa aagagccgtt catttcaata aaccgggcga
     4741 cctcagccat cccttcctga ttttccgctt tccagcgttc ggcacgcaga cgacgggctt
     4801 cattctgcat ggttgtgctt accagaccgg agatattgac atcatatatg ccttgagcaa
     4861 ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat acctcttttt
     4921 gacatacttc gggtatacat atcagtatat attcttatac cgcaaaaatc agcgcgcaaa
     4981 tacgcatact gttatctggc ttttagtaag ccggatccag atctttacgc cccgccctgc
     5041 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     5101 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     5161 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     5221 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     5281 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     5341 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     5401 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     5461 atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     5521 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     5581 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     5641 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     5701 cctgaaaatc tcgacggatc ctaactcaaa atccacacat tatacgagcc ggaagcataa
     5761 agtgtaaagc ctggggtgcc taatgcggcc gccatagtga ctggatatgt tgtgttttac
     5821 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     5881 tttacgtttc tcgttcagct tttttgtaca aacttgttga tacgcgtggt acatcaatgg
     5941 gatcatatct tgctgcctcc cgcagctcac cgtgcgagct gccgggaccc cgtttctcat
     6001 aggcgtgtgc accttccgtc tgagacctgt aaatggttaa aaaggaatat ataattacct
     6061 tttgaaaagt ggtaaaaaac gaatatatct ttttttttgg ttttaatacg ctcttttgtg
     6121 tgtatgagag gaataaataa gtgtgtgtgt gtgtgtgtgt tgttgttgtt gaaaaaatga
     6181 aggacacgaa agcgctcatt aggggtgggg atacaccagg cataattatc cgaaaatatt
     6241 ttgatgcacc aaataagtga acatactgac gaaatcaaag agttggagga tggataggga
     6301 ggcctcattg gcagtggtaa attgatttga cttgaatatg ataccctctt actgtttcat
     6361 tgccactaca agttggtttc cttcccctgc aggcatgcaa gctagcttgt attctatagt
     6421 gtcacctaaa tcgtatgtgt atgatacata aggttatgta ttaattgtag ccgcgttcta
     6481 acgacaatat gtacaagcct aattgtgtag catctggctt actgaagcag accctatcat
     6541 ctctctcgta aactgccgtc agagtcggtt tggttggacg aaccttctga gtttctggta
     6601 acgccgttcc gcaccccgga aatggtcagc gaaccaatca gcagggtcat cgctagccag
     6661 atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc
     6721 gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc
     6781 gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc
     6841 tccttgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct
     6901 tcctaatgca ggagtcgcat aagggagagc gtcgatatgg tgcactctca gtacaatctg
     6961 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     7021 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     7081 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     7141 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     7201 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     7261 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     7321 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     7381 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     7441 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     7501 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     7561 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     7621 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     7681 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     7741 cggaggaccg aaggagctaa ccgctttttt gcacagcatg ggggatcatg taactcgcct
     7801 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     7861 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7921 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7981 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     8041 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     8101 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     8161 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     8221 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     8281 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     8341 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     8401 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     8461 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     8521 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     8581 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     8641 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     8701 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     8761 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     8821 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     8881 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     8941 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     9001 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     9061 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     9121 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcag
//
