LOCUS       sequence-68943-e        7686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Trasgenically expresses tandem tdtomato fluorescent protein in T. 
            cruzi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7686)
  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 7686)
  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-Neo-tdTomato
FEATURES             Location/Qualifiers
     source          1..7686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(33..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1724..3148
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMN
                     FEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVL
                     KGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYER
                     SEGRHHLFLYGMDELYK"
     CDS             3923..4726
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3986..4005)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4596..4615
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     promoter        complement(5501..5519)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5540..5556)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5540..5556)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5553..5575)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5564..5580
                     /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(5588..5594,5595..5612,5613..5618))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5633..5654
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5771..5788)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5942..6530)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6022..6041)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6701..7492,7493..7561))
                     /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     7324..7343
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7562..7666)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcgg
      661 tgcaccctgg gatatttgcg cacccacctt tcccgtcgca atcggcgccg tccgaacgcg
      721 gaaatgttcc gagaataata cggctgggct gcgccgcgcg ttccccgcgt gaaccaacta
      781 ttgtgcgatg tgcagcatcc tattgtggtg ctttaccgca caccagtttc cttgcggggc
      841 ttatttggtg tgttggattt ttaatttttt gatgattcgg aggaattttt tgcaattttt
      901 gggtacccct aaaaaaattt tggagtttcc ggggttcgat ccaacttgtg cttttttaac
      961 cgtccatttg tgtctagtac atcatggagc ggtattctct ccggtggccc aggtggcgtc
     1021 ggaaggaatc ggtgtccctc aggatgtttg gtttttcgtt tttttttttg cggtcttttg
     1081 tttcgcatcg ccgtatcgtg tgcggcgcga ccttcttttt ggctcattgg tcagtgaggt
     1141 ttataataaa gtggatcatt tgggtttttg gccctttgcg gcgcacccgc ggaatggggg
     1201 agggattttc atataattgt agtgtacaca ttgtgtgtgg gtcgcaaaat gtcatgccgt
     1261 gtgcaatgtt tatttgcgtg ctgtgcgtgt ccttttgtga ttccaccact tgtgtgtatt
     1321 ttaccttatc gtctctccct cgcgttggct gtgcgtgaat tgtctgtcgc attcgtggga
     1381 aatcaaaagg ggaatcaacg tgttcttttt ttttcagaag gggagtcaaa gcgccgtgtg
     1441 gatgccaagt ctgtgcaaga gttctagtgg atcccccaac gagtttcttc aaaatatgca
     1501 gcggattcac taagaaacat tttcacgcac gaaagcgaaa ttattatgat tgttattata
     1561 atactttttc tttgttgttt tatccactta ttattgttgt gttaaattgt ttttaccttt
     1621 tttcttttcc aacttctttt atgatgtctt ttcttttttt tttttgctct ataagttgtc
     1681 ttgtctagaa ttccctttta gatctgctag catggcgcct agggtgagca agggcgagga
     1741 ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga
     1801 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     1861 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat
     1921 gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc
     1981 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac
     2041 cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg
     2101 caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     2161 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa
     2221 gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc
     2281 cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct cccacaacga
     2341 ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctggg
     2401 gcatggcacc ggcagcaccg gcagcggcag ctccggcacc gcctcctccg aggacaacaa
     2461 catggccgtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg
     2521 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     2581 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     2641 gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa
     2701 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct
     2761 ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat
     2821 gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga
     2881 ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc
     2941 cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa
     3001 gaagcccgtg caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca
     3061 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt
     3121 cctgtacggc atggacgagc tgtacaagcc cggggacgtc gatatcgagc tctaagtcga
     3181 cctcgagcca tttacgactc caaggcaacg ctgcagaaca acctgccgaa ggagcgccgc
     3241 ttcttcaaga ttgtgtcgtg gtacgacaac gagtggggat actcccaccg cgtggtggac
     3301 cttgtacgcc acatggcctc gaaggatcgt tcggcaaggt tgtaggcgtg gcgatgactt
     3361 caggtctttc ttttgcgaat agggatctta taatacacga tgcgtgtccc gtgatgatcg
     3421 ttaccggtgc tgccacgatc caattgacac agcgtcaaga gcaaaacaat tttacttttc
     3481 cctttaagga caacaacaaa aaaatatata actttttttt cttttttttt ttttttgaaa
     3541 ttatatttat ggtcatcttt gggaaacaaa aagcagcaat ttaatgatgc ggaaggatga
     3601 gtgaaataat gtttaatcaa tgtacgagga tttggggtat tgcaaggaaa atgtagatga
     3661 tttaattggg tgtgtgatgc agcttgtggt aatttttgct cacttccctt tttgccacat
     3721 ctttttagtt tttctgcttt tctttcccca ttattccact tgtctctctt ttcccacgtt
     3781 tcctgcacga atgcagaaag tgatattttt actttgaaag ccatctacca acaacaatta
     3841 cattgaacag aatttgggaa ttgcgaatta attcctgcag ccccgcgtgt ggcctcgaac
     3901 accgagcgac cctgcagcca atatgggatc ggccattgaa caagatggat tgcacgcagg
     3961 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4021 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4081 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4141 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4201 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4261 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4321 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4381 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4441 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4501 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4561 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4621 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4681 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagggg atcgatccgg
     4741 aacaacctgc cgaaggagcg ccgcttcttc aagattgtgt cgtggtacga caacgagtgg
     4801 ggatactccc accgcgtggt ggaccttgta cgccacatgg cctcgaagga tcgttcggca
     4861 aggttgtagg cgtggcgatg acttcaggtc tttcttttgc gaatagggat cttataatac
     4921 acgatgcgtg tcccgtgatg atcgttaccg gtgctgccac gatccaagtg acacagtgtc
     4981 aagagcaaaa caattttact tttcccttta aggacaacaa caaaaaaata tataactttt
     5041 ttttcttttt ttttttttga aattatattt atggtcatct ttgggaaaca aaaagcagca
     5101 atttaatgat gcggaaggat gagtgaaata atgtttaatc aatgtacgag gatttggggt
     5161 attgcaagga aaatgtagat gatttagttg ggtgtatgat gcagcttgtg gtaatttttt
     5221 acaactttgc gttattggaa gattttggtt gttcagtttc tttttgttta taactgactg
     5281 ttttaattta gtgtgatttt ttttttttaa ggatttaagc tgtttttctt tttttttctt
     5341 tttcttcact gatggtgttt aaggtgtagt ttagtgtttg tgcattatgc gtcgtagtgt
     5401 tgttgttctt tggacttttt cgaacaatac tgggcattat agcccacctg tgtatatgcc
     5461 gctagagtat gctagccgtg tgggtaccca gcttttgttc cctttagtga gggttaattt
     5521 cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5581 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5641 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5701 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5761 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5821 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5881 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5941 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6001 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6061 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6121 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6181 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6241 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6301 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6361 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6421 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6481 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6541 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6601 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6661 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6721 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6781 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6841 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6901 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6961 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7021 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7081 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7141 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7201 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7261 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7321 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7381 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7441 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7501 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7561 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7621 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7681 aagtgc
//
