LOCUS       sequence-69267-e        7954 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Transgenically expresses firefly luciferase protein in T. cruzi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7954)
  AUTHORS   Canavaci AM, Bustamante JM, Padilla AM, Perez Brandan CM, Simpson 
            LJ, Xu D, Boehlke CL, Tarleton RL
  TITLE     In vitro and in vivo high-throughput assays for the testing of 
            anti-Trypanosoma cruzi compounds.
  JOURNAL   PLoS Negl Trop Dis. 2010 Jul 13;4(7):e740. doi: 
            10.1371/journal.pntd.0000740.
  PUBMED    20644616
REFERENCE   2  (bases 1 to 7954)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS Negl 
            Trop Dis."; date: "2010-07-13"; volume: "4(7):e740. doi"; pages: " 
            10.1371/journal.pntd.0000740"
COMMENT     Sequence Label: pLew90-beta-GW/T7/PARP SAS/luciferase/Aldolase 3′UTR
FEATURES             Location/Qualifiers
     source          1..7954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     703..719
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     703..719
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    739..759
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        768..786
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    884..908
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             914..2566
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(980..998)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    complement(2568..2592)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2755..2775)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     terminator      2849..2896
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2981..3028
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4195..4214)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4805..4824
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5430..5449)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5660..5679)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5839..5857)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5925..6029
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6030..6098,6099..6890)
                     /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(6248..6267)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7061..7649
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7550..7569
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7803..7820
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        7901..7919
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atgaagccct gtacgatatt tgttttcgta cgttgaaact gacgactcca acgttcggtg
       61 atctgaacca cttggtgtcc gcggtggttt ccggcgtgac ctgctgcctg cgcttccctg
      121 gccagctcaa ctccgacctg cgcaagctgg cggtgaacct ggtgccgttc cctcgtctgc
      181 acttcttcat gatgggcttt gccccgctca gcagccgcgg ctcgcagcag taccgaggtc
      241 tgtccgtgcc ggacgtgacg cagcagatgt tcgatgcaaa gaacatgatg caggctgcag
      301 acccggccca cggtcgctac ctgacggcgt ctgcgctctt ccgcggccgc atgtcgacga
      361 aggaggtgga cgagcagatg ctcaacgtgc agaacaagaa ctcgtcctac ttcattgagt
      421 ggatcccgaa caacatcaag tcctccatct gcgacatccc gcccaagggc ctcaagatgg
      481 ccgtcacctt tgtcggcaac aacacctgca tccaggagat gtttcgccgc gtgggcgagc
      541 agttcacggc gatgttccgc cgcaaggcgt tcttgcactg gtacacgggc gagggaatgg
      601 acgagatgga gttcaccgag gcggagtcca acatgaacga tcttgtgtcg gagtaccagc
      661 agtaccagga cgccaccatt gaggaggaag ctcgagaccg gtcaggaaac agctatgacc
      721 atgattacgc caagctatca actttgtata gaaaagttgg gagctcgtaa tacgactcac
      781 tatagggcga attggatcct gcacgcgcct tcgagttttt tttccttttc cccatttttt
      841 tcaacttgaa gacttcaatt acaccaaaaa gtaaaattca caagcaagtt tgtacaaaaa
      901 agcaggctca accatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc
      961 tctagaggat ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt
     1021 tcctggaaca attgctttta cagatgcaca tatcgaggtg aacatcacgt acgcggaata
     1081 cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca
     1141 cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt
     1201 atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa
     1261 cagtatgaac atttcgcagc ctaccgtagt gtttgtttcc aaaaaggggt tgcaaaaaat
     1321 tttgaacgtg caaaaaaaat taccaataat ccagaaaatt attatcatgg attctaaaac
     1381 ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt
     1441 taatgaatac gattttgtac cagagtcctt tgatcgtgac aaaacaattg cactgataat
     1501 gaattcctct ggatctactg ggttacctaa gggtgtggcc cttccgcata gaactgcctg
     1561 cgtcagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc
     1621 gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt
     1681 gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttttacgatc
     1741 ccttcaggat tacaaaattc aaagtgcgtt gctagtacca accctatttt cattcttcgc
     1801 caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg cttctggggg
     1861 cgcacctctt tcgaaagaag tcggggaagc ggttgcaaaa cgcttccatc ttccagggat
     1921 acgacaagga tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga
     1981 tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct
     2041 ggataccggg aaaacgctgg gcgttaatca gagaggcgaa ttatgtgtca gaggacctat
     2101 gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg
     2161 atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct tcatagttga
     2221 ccgcttgaag tctttaatta aatacaaagg atatcaggtg gcccccgctg aattggaatc
     2281 gatattgtta caacacccca acatcttcga cgcgggcgtg gcaggtcttc ccgacgatga
     2341 cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa
     2401 agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt
     2461 tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag
     2521 agagatcctc ataaaggcca agaagggcgg aaagatcgcc gtgtaagacc cagctttctt
     2581 gtacaaagtg gggtcttaag gatcctgccc atttggcttt tcccttgtct cgtgtctttt
     2641 ccgtggaaag gttcccggag taatctgatg gcacagcagg gaggtgcgcc tgcaggttgg
     2701 ttaggaaggg gggatgatgt aaaagaagaa aatgggggga ttcgagcccg ggcacaactt
     2761 tattatacat agttgataat tcactggccg tgggcccttc gaaggtaccg agctcttaat
     2821 taaaagcttc cgagccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     2881 cttgaggggt tttttgctga aaggagaact atatccggat agggctgcta acaaagcccg
     2941 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     3001 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gctacgcgtt
     3061 ctagacccgg ggatatcact agtgatgaaa aagcctgaac tcaccgcgac gtctgtcgag
     3121 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     3181 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     3241 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     3301 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc
     3361 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     3421 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     3481 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatga
     3541 gtcttgtgat gtactggctg atttctacga ccagttcgct gaccagttgc acgagtctca
     3601 attggacaaa atgccagcac ttccggctaa aggtaacttg aacctccgtg acatcttaga
     3661 gtcggacttc gcgttcgcgt aacgccaaat caatacgact cactatagag ggacaaactc
     3721 aaggtcattc gcaagagtgg ccggatcctg cccatttagt tagttggctt ttcccttgtc
     3781 tcgtgtcttt tccgtggaaa ggttcccgga gtaatctgat ggcacagcag ggaggtgcgc
     3841 ctgcaggttg gttaggaagg ggggatgatg taaaagaaga aaatgggggg attcgagccc
     3901 gggcacagca aggtcttctg aaattcatgt tttttttttt tttactctgc attgcagtct
     3961 ccgctcttat ttagttttgc tttacgtaag gtctcgttgc tgccataaaa taagcttaga
     4021 ggagaaagaa tagtaaccct ttcatcaaag aaaatagttc aaacgaatat gcgcgaaatc
     4081 gtctgcgttc aggctggcca atgcggtaac cagatcggct caaagttctg ggaggtgatc
     4141 cggccaagct tggatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4201 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4261 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4321 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4381 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4441 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4501 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4561 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4621 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4681 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4741 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4801 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4861 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4921 gacgagttct tctgagcggg actctggggt tcggatccta acaccgggtt gtgtggccaa
     4981 aattgttctg tagtcgctgt gagttgacac ggctagtgct tatgattttc ctcgcgtgtg
     5041 gtgcctgtac tcagccctat gccttatttg caacacattt acgtacagcg cacaagagaa
     5101 gagaagatca cttgaagata ataaatatag ggttgtaggc atcttgttta actcaaattt
     5161 tctcgtcttg gtgtgtcgac atgattgaaa tagtgccacc agttgtgttt gatgcgtttg
     5221 ttatctatgc agtattctgc aagggggcac agcaaggtct tctgaaattc atgttttttt
     5281 tttttttact ctgcattgca gtctccgctc ttatttagtt ttgctttacg taaggtctcg
     5341 ttgctgccat aaaataagct ctagccagat cctctacgcc ggacgcatcg tggccggcat
     5401 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
     5461 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
     5521 cgtggccggg ggactgttgg gcgccatctc cttgcaccat tccttgcggc ggcggtgctc
     5581 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt
     5641 cgatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     5701 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     5761 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     5821 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     5881 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5941 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6001 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6061 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6121 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6181 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6241 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6301 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6361 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6421 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6481 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6541 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6601 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6661 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6721 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6781 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6841 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6901 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6961 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7021 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7081 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7141 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7201 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7261 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7321 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7381 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7441 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7501 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7561 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7621 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7681 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7741 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7801 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     7861 cgcgttggcc gattcattaa tgcagctggc ttatcgaaat taatacgact cactataggg
     7921 agaccggaat tcgagctcgg tacctgtatt gaca
//
