LOCUS       sequence-176691-        6092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  [Edit] Expresses recombinant 6HIS tagged T. cruzi mitochondrial 
            tryparedoxin peroxidase protein in E. coli upon IPTG induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6092)
  AUTHORS   Dotiwala F, Mulik S, Polidoro RB, Ansara JA, Burleigh BA, Walch M, 
            Gazzinelli RT, Lieberman J
  TITLE     Killer lymphocytes use granulysin, perforin and granzymes to kill 
            intracellular parasites.
  JOURNAL   Nat Med. 2016 Feb;22(2):210-6. doi: 10.1038/nm.4023. Epub 2016 Jan 
            11.
  PUBMED    26752517
REFERENCE   2  (bases 1 to 6092)
  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"; doi: "10.1038/nm.4023"; 
            journalName: "Nat Med"; date: "2016-02"; volume: "22"; issue: "2"; 
            pages: "210-6"
COMMENT     Sequence Label: pET21a - MPX
FEATURES             Location/Qualifiers
     source          1..6092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..33
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             736..753
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(806..824)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      820..867
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      904..1359
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(991..1010)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1201..1222
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1385..1489
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1490..1558,1559..2350)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1708..1727)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2521..3109
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3010..3029
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3263..3280
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3295..3434
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3360..3379)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3520..3542
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3536..3727)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4536..5618)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5580..5599
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5619..5696)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5902..5921
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6005..6024
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6005..6023
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6024..6048
                     /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)."
     RBS             6063..6085
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atggctagca tgactggtgg acagcaaatg ggtcgcggat ccatgtttcg tcgtatggcc
       61 gtgacttcgt tgcagaaggg tctttcacgc cgggctttct gcaacaccct gcgacttctt
      121 aacctggact accaagcata taagactgcc acggtgcgcg aggcagcacc tgagtgggca
      181 ggaaaagctg tcgtgaatgg aaaaatccag gacatcagct tgaatgacta caagggcaag
      241 tatgttgttc ttctttttta tcctatggat tttacctttg tgtgccccac ggaaatcaca
      301 gccttttcag atgctcaggc agaatttgat aagattaaca cccaggtcgt ggctgtatca
      361 tgcgacagcc aatattcaca ccttgcttgg ataaacactc caagaaataa aggtggcctt
      421 ggggaaatga gtattcctgt gctttctgat ctcaccaagg aaattgcccg cgactatgga
      481 gttttgatcg aggagcaagg gatctctctg cgcggactgt tcatcattga cgataaaggg
      541 attctgcgcc acatcacggt gaatgacttg ccagtgggca gaaatgttga agaggtgctt
      601 cgggttgtgc aggcgtttca gtatgtggac aaaaacggag atgtaattcc atgtaactgg
      661 aggccaggca aaccgacaat gaaaactgag aaggcaaatg aatattttga gaaaaacgca
      721 gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
      781 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
      841 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
      901 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      961 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1021 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1081 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1141 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1201 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     1261 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     1321 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     1381 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1441 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1501 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1561 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1621 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1681 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1741 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1801 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1861 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1921 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1981 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     2041 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2101 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2161 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2221 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2281 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2341 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2401 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2461 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2521 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2581 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2641 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2701 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2761 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2821 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2881 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2941 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3001 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3061 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3121 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3181 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3241 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     3301 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
     3361 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     3421 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3481 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3541 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3601 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3661 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     3721 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     3781 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     3841 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     3901 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     3961 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     4021 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     4081 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     4141 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg
     4201 ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga
     4261 aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg
     4321 cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta
     4381 cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc
     4441 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
     4501 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4561 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4621 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
     4681 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
     4741 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
     4801 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
     4861 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
     4921 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
     4981 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
     5041 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
     5101 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
     5161 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
     5221 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
     5281 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     5341 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     5401 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     5461 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     5521 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     5581 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     5641 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     5701 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     5761 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     5821 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     5881 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     5941 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
     6001 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
     6061 attttgttta actttaagaa ggagatatac at
//
