LOCUS       sequence-109234-        7483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Trasgenically expresses firefly luciferase in T. cruzi with 
            phleomycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7483)
  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 7483)
  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: "PLoS Negl 
            Trop Dis."; date: "2010-07-13"; volume: "4(7):e740. doi"; pages: " 
            10.1371/journal.pntd.0000740"
COMMENT     Sequence Label: pTrex-Luc-Phleo
FEATURES             Location/Qualifiers
     source          1..7483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             721..1095
                     /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(1931..1949)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1970..1986)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1970..1986)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1983..2005)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1994..2010
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2018..2024,2025..2042,2043..2048))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2063..2084
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2201..2218)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2372..2960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2452..2471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3131..3922,3923..3991))
                     /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     3754..3773
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3992..4096)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4122..4577)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4259..4280)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4471..4490
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4704..4726
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4718..4735
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4719..4735
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4742..4761
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4742..4760
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5908..5926)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
ORIGIN
        1 tcgagccatt tacgactcca aggcaacgct gcagaacaac ctgccgaagg agcgccgctt
       61 cttcaagatt gtgtcgtggt acgacaacga gtggggatac tcccaccgcg tggtggacct
      121 tgtacgccac atggcctcga aggatcgttc ggcaaggttg taggcgtggc gatgacttca
      181 ggtctttctt ttgcgaatag ggatcttata atacacgatg cgtgtcccgt gatgatcgtt
      241 accggtgctg ccacgatcca attgacacag cgtcaagagc aaaacaattt tacttttccc
      301 tttaaggaca acaacaaaaa aatatataac ttttttttct tttttttttt ttttgaaatt
      361 atatttatgg tcatctttgg gaaacaaaaa gcagcaattt aatgatgcgg aaggatgagt
      421 gaaataatgt ttaatcaatg tacgaggatt tggggtattg caaggaaaat gtagatgatt
      481 taattgggtg tgtgatgcag cttgtggtaa tttttgctca cttccctttt tgccacatct
      541 ttttagtttt tctgcttttc tttccccatt attccacttg tctctctttt cccacgtttc
      601 ctgcacgaat gcagaaagtg atatttttac tttgaaagcc atctaccaac aacaattaca
      661 ttgaacagaa tttgggattg cgagatctag atatcgataa tattctagct gagggtaccc
      721 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
      781 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
      841 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
      901 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
      961 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     1021 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     1081 gaggagcagg actgaccgac gccgaccaac accgccggtc cgacggcggc ccacgggtcc
     1141 caggcctcgg agatctgggg atcgatccgg aacaacctgc cgaaggagcg ccgcttcttc
     1201 aagattgtgt cgtggtacga caacgagtgg ggatactccc accgcgtggt ggaccttgta
     1261 cgccacatgg cctcgaagga tcgttcggca aggttgtagg cgtggcgatg acttcaggtc
     1321 tttcttttgc gaatagggat cttataatac acgatgcgtg tcccgtgatg atcgttaccg
     1381 gtgctgccac gatccaagtg acacagtgtc aagagcaaaa caattttact tttcccttta
     1441 aggacaacaa caaaaaaata tataactttt ttttcttttt ttttttttga aattatattt
     1501 atggtcatct ttgggaaaca aaaagcagca atttaatgat gcggaaggat gagtgaaata
     1561 atgtttaatc aatgtacgag gatttggggt attgcaagga aaatgtagat gatttagttg
     1621 ggtgtatgat gcagcttgtg gtaatttttt acaactttgc gttattggaa gattttggtt
     1681 gttcagtttc tttttgttta taactgactg ttttaattta gtgtgatttt ttttttttaa
     1741 ggatttaagc tgtttttctt tttttttctt tttcttcact gatggtgttt aaggtgtagt
     1801 ttagtgtttg tgcattatgc gtcgtagtgt tgttgttctt tggacttttt cgaacaatac
     1861 tgggcattat agcccacctg tgtatatgcc gctagagtat gctagccgtg tgggtaccca
     1921 gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt
     1981 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2041 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2101 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2161 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2221 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2281 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2341 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2401 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2461 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2521 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2581 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2641 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2701 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2761 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     2821 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2881 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2941 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3001 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3061 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3121 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3181 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3241 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3301 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3361 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3421 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3481 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3541 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3601 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3661 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3721 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3781 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3841 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3901 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3961 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4021 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4081 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4141 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4201 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4261 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4321 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4381 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4441 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4501 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4561 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4621 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     4681 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     4741 gtaatacgac tcactatagg gcgaattgga gctcggtgca ccctgggata tttgcgcacc
     4801 cacctttccc gtcgcaatcg gcgccgtccg aacgcggaaa tgttccgaga ataatacggc
     4861 tgggctgcgc cgcgcgttcc ccgcgtgaac caactattgt gcgatgtgca gcatcctatt
     4921 gtggtgcttt accgcacacc agtttccttg cggggcttat ttggtgtgtt ggatttttaa
     4981 ttttttgatg attcggagga attttttgca atttttgggt acccctaaaa aaattttgga
     5041 gtttccgggg ttcgatccaa cttgtgcttt tttaaccgtc catttgtgtc tagtacatca
     5101 tggagcggta ttctctccgg tggcccaggt ggcgtcggaa ggaatcggtg tccctcagga
     5161 tgtttggttt ttcgtttttt tttttgcggt cttttgtttc gcatcgccgt atcgtgtgcg
     5221 gcgcgacctt ctttttggct cattggtcag tgaggtttat aataaagtgg atcatttggg
     5281 tttttggccc tttgcggcgc acccgcggaa tgggggaggg attttcatat aattgtagtg
     5341 tacacattgt gtgtgggtcg caaaatgtca tgccgtgtgc aatgtttatt tgcgtgctgt
     5401 gcgtgtcctt ttgtgattcc accacttgtg tgtattttac cttatcgtct ctccctgccg
     5461 ttggctgtgc gtgaattgtg cttcgcattc gtgggaaatc aaaaggggaa tcaacgtgtt
     5521 cttttttttt cagaagggga gtcaaagcgc cgtgtggatg ccaagtctgt gcaagagttc
     5581 tagtggatcc cccaacgagt ttcttcaaaa tatgcagcgg attcactaag aaacattttc
     5641 acgcacgaaa gcgaaattat tatgattgtt attataatac tttttctttg ttgttttatc
     5701 cacttattat tgttgtgtta aattgttttt accttttttc ttttccaact tcttttatga
     5761 tgtcttttct tttttttttt tttgctctat aagttgtctt gtctagaatt cgatatcaag
     5821 cttggtaccg agctcggatc catggaagac gccaaaaaca taaagaaagg cccggcgcca
     5881 ttctatcctc tagaggatgg aaccgctgga gagcaactgc ataaggctat gaagagatac
     5941 gccctggttc ctggaacaat tgcttttaca gatgcacata tcgaggtgaa catcacgtac
     6001 gcggaatact tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata tgggctgaat
     6061 acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc aattctttat gccggtgttg
     6121 ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa
     6181 ttgctcaaca gtatgaacat ttcgcagcct accgtagtgt ttgtttccaa aaaggggttg
     6241 caaaaaattt tgaacgtgca aaaaaaatta ccaataattc agaaaattat tatcatggat
     6301 tctaaaacgg attaccaggg atttcagtcg atgtacacgt tcgtcacatc tcatctacct
     6361 cccggtttta atgagtacga ttttgtacca gagtcctttg atcgtgacaa aacaattgca
     6421 ctgataatga actcctctgg atctactggg ttacctaagg gtgtggccct tccgcataga
     6481 actgcctgcg tcagattctc gcatgccaga gatcctattt ttggcaatca aatcattccg
     6541 gatactgcga ttttaagtgt tgttccattc catcacggtt ttggaatgtt tactacactc
     6601 ggatatttga tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga agagctgttt
     6661 ttacgatccc ttcaggatta caaaattcaa agtgcgttgc tagtaccaac cctattttca
     6721 ttcttcgcca aaagcactct gattgacaaa tacgatttat ctaatttaca cgaaattgct
     6781 tctgggggcg cacctctttc gaaagaagtc ggggaagcgg ttgcaaaacg cttccatctt
     6841 ccagggatac gacaaggata tgggctcact gagactacat cagctattct gattacaccc
     6901 gagggggatg ataaaccggg cgcggtcggt aaagttgttc cattttttga agcgaaggtt
     6961 gtggatctgg ataccgggaa aacgctgggc gttaatcaga gaggcgaatt atgtgtcaga
     7021 ggacctatga ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc cttgattgac
     7081 aaggatggat ggctacattc tggagacata gcttactggg acgaagacga acacttcttc
     7141 atagttgacc gcttgaagtc tttaattaaa tacaaaggat atcaggtggc ccccgctgaa
     7201 ttggaatcga tattgttaca acaccccaac atcttcgacg cgggcgtggc aggtcttccc
     7261 gacgatgacg ccggtgaact tcccgccgcc gttgttgttt tggagcacgg aaagacgatg
     7321 acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa aaagttgcgc
     7381 ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga
     7441 aaaatcagag agatcctcat aaaggccaag aagggcggaa agc
//
