LOCUS       sequence-118938-        7957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Trasgenically expresses firefly luciferase in T. cruzi with neomycin
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7957)
  AUTHORS   Sanchez-Valdez FJ, Padilla A, Wang W, Orr D, Tarleton RL
  TITLE     Spontaneous dormancy protects Trypanosoma cruzi during extended drug
            exposure.
  JOURNAL   Elife. 2018 Mar 26;7. pii: 34039. doi: 10.7554/eLife.34039.
  PUBMED    29578409
REFERENCE   2  (bases 1 to 7957)
  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: "Elife."; 
            date: "2018-03-26"; volume: "7. pii: 34039. doi"; pages: " 
            10.7554/eLife.34039"
COMMENT     Sequence Label: pTrex-Luc-Neo
FEATURES             Location/Qualifiers
     source          1..7957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(977..995)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             3293..4096
                     /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(3356..3375)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3966..3985
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     promoter        complement(4959..4977)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4998..5014)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4998..5014)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5011..5033)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5022..5038
                     /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(5046..5052,5053..5070,5071..5076))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5091..5112
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5229..5246)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5400..5988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5480..5499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6159..6950,6951..7019))
                     /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     6782..6801
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7020..7124)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7150..7605)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7287..7308)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7499..7518
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7732..7754
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7746..7763
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7747..7763
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7770..7789
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7770..7788
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 taccgcacac cagtttcctt gcggggctta tttggtgtgt tggattttta attttttgat
       61 gattcggagg aattttttgc aatttttggg tacccctaaa aaaattttgg agtttccggg
      121 gttcgatcca acttgtgctt ttttaaccgt ccatttgtgt ctagtacatc atggagcggt
      181 attctctccg gtggcccagg tggcgtcgga aggaatcggt gtccctcagg atgtttggtt
      241 tttcgttttt ttttttgcgg tcttttgttt cgcatcgccg tatcgtgtgc ggcgcgacct
      301 tctttttggc tcattggtca gtgaggttta taataaagtg gatcatttgg gtttttggcc
      361 ctttgcggcg cacccgcgga atgggggagg gattttcata taattgtagt gtacacattg
      421 tgtgtgggtc gcaaaatgtc atgccgtgtg caatgtttat ttgcgtgctg tgcgtgtcct
      481 tttgtgattc caccacttgt gtgtatttta ccttatcgtc tctccctgcc gttggctgtg
      541 cgtgaattgt gcttcgcatt cgtgggaaat caaaagggga atcaacgtgt tctttttttt
      601 tcagaagggg agtcaaagcg ccgtgtggat gccaagtctg tgcaagagtt ctagtggatc
      661 ccccaacgag tttcttcaaa atatgcagcg gattcactaa gaaacatttt cacgcacgaa
      721 agcgaaatta ttatgattgt tattataata ctttttcttt gttgttttat ccacttatta
      781 ttgttgtgtt aaattgtttt tacctttttt cttttccaac ttcttttatg atgtcttttc
      841 tttttttttt ttgctctata agttgtcttg tctagaattc gatatcaagc ttggtaccga
      901 gctcggatcc atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct
      961 ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
     1021 tggaacaatt gcttttacag atgcacatat cgaggtggac atcacgtacg cggaatactt
     1081 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
     1141 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
     1201 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
     1261 tatgaacatt tcgcagccta ccgtagtgtt tgtttccaaa aaggggttgc aaaaaatttt
     1321 gaacgtgcaa aaaaaattac caataattca gaaaattatt atcatggatt ctaaaacgga
     1381 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
     1441 tgagtacgat tttgtaccag agtcctttga tcgtgacaaa acaattgcac tgataatgaa
     1501 ctcctctgga tctactgggt tacctaaggg tgtggccctt ccgcatagaa ctgcctgcgt
     1561 cagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
     1621 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
     1681 atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttt tacgatccct
     1741 tcaggattac aaaattcaaa gtgcgttgct agtaccaacc ctattttcat tcttcgccaa
     1801 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctgggggcgc
     1861 acctctttcg aaagaagtcg gggaagcggt tgcaaaacgc ttccatcttc cagggatacg
     1921 acaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
     1981 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
     2041 taccgggaaa acgctgggcg ttaatcagag aggcgaatta tgtgtcagag gacctatgat
     2101 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
     2161 gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca tagttgaccg
     2221 cttgaagtct ttaattaaat acaaaggata tcaggtggcc cccgctgaat tggaatcgat
     2281 attgttacaa caccccaaca tcttcgacgc gggcgtggca ggtcttcccg acgatgacgc
     2341 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
     2401 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
     2461 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
     2521 gatcctcata aaggccaaga agggcggaaa gctcgagcca tttacgactc caaggcaacg
     2581 ctgcagaaca acctgccgaa ggagcgccgc ttcttcaaga ttgtgtcgtg gtacgacaac
     2641 gagtggggat actcccaccg cgtggtggac cttgtacgcc acatggcctc gaaggatcgt
     2701 tcggcaaggt tgtaggcgtg gcgatgactt caggtctttc ttttgcgaat agggatctta
     2761 taatacacga tgcgtgtccc gtgatgatcg ttaccggtgc tgccacgatc caattgacac
     2821 agcgtcaaga gcaaaacaat tttacttttc cctttaagga caacaacaaa aaaatatata
     2881 actttttttt cttttttttt ttttttgaaa ttatatttat ggtcatcttt gggaaacaaa
     2941 aagcagcaat ttaatgatgc ggaaggatga gtgaaataat gtttaatcaa tgtacgagga
     3001 tttggggtat tgcaaggaaa atgtagatga tttaattggg tgtgtgatgc agcttgtggt
     3061 aatttttgct cacttccctt tttgccacat ctttttagtt tttctgcttt tctttcccca
     3121 ttattccact tgtctctctt ttcccacgtt tcctgcacga atgcagaaag tgatattttt
     3181 actttgaaag ccatctacca acaacaatta cattgaacag aatttgggaa ttgcgaatta
     3241 attcctgcag ccccgcgtgt ggcctcgaac accgagcgac cctgcagcca atatgggatc
     3301 ggccattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3361 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3421 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3481 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3541 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3601 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3661 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3721 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3781 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     3841 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3901 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3961 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4021 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4081 tgacgagttc ttctgagggg atcgatccgt gcacaggcgt tagccactct caggattgcc
     4141 agcaagtctt ctgaatttga tctttggatg ctcgggcagc tcaaagagga atcttagtca
     4201 caacctgccg aaggagcgcc gcttcttcaa gattgtgtcg tggtacgaca acgagtgggg
     4261 atactcccac cgcgtggtgg accttgtacg ccacatggcc tcgaaggatc gttcggcaag
     4321 gttgtaggcg tggcgatgac ttcaggtctt tcttttgcga atagggatct tataatacac
     4381 gatgcgtgtc ccgtgatgat cgttaccggt gctgccacga tccaagtgac acagtgtcaa
     4441 gagcaaaaca attttacttt tccctttaag gacaacaaca aaaaaatata taactttttt
     4501 ttcttttttt ttttttgaaa ttatatttat ggtcatcttt gggaaacaaa aagcagcaat
     4561 ttaatgatgc ggaaggatga gtgaaataat gtttaatcaa tgtacgagga tttggggtat
     4621 tgcaaggaaa atgtagatga tttagttggg tgtatgatgc agcttgtggt aattttttac
     4681 aactttgcgt tattggaaga ttttggttgt tcagtttctt tttgtttata actgactgtt
     4741 ttaatttagt gtgatttttt ttttttaagg atttaagctg tttttctttt tttttctttt
     4801 tcttcactga tggtgtttaa ggtgtagttt agtgtttgtg cattatgcgt cgtagtgttg
     4861 ttgttctttg gactttttcg aacaatactg ggcattatag cccacctgtg tatatgccgc
     4921 tagagtatgc tagccgtgtg ggtacccagc ttttgttccc tttagtgagg gttaatttcg
     4981 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5041 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5101 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5161 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5221 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5281 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5341 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5401 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5461 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5521 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5581 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5641 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5701 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5761 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5821 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5881 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5941 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6001 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6061 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6121 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6181 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6241 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6301 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6361 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6421 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6481 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6541 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6601 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6661 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6721 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6781 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6841 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6901 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6961 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7021 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7081 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7141 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     7201 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     7261 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     7321 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     7381 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     7441 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     7501 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     7561 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     7621 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     7681 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     7741 gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc gaattggagc
     7801 tcggtgcacc ctgggatatt tgcgcaccca cctttcccgt cgcaatcggc gccgtccgaa
     7861 cgcggaaatg ttccgagaat aatacggctg ggctgcgccg cgcgttcccc gcgtgaacca
     7921 actattgtgc gatgtgcagc atcctattgt ggtgctt
//
