LOCUS       sequence-176693-        6113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses recombinant 6HIS tagged T. cruzi superoxide dismutase A 
            protein in E. coli upon IPTG induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6113)
  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 6113)
  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 - SodA
FEATURES             Location/Qualifiers
     source          1..6113
                     /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             757..774
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(827..845)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      841..888
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      925..1380
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1012..1031)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1222..1243
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1406..1510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1511..1579,1580..2371)
                     /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(1729..1748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2542..3130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3031..3050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3284..3301
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3316..3455
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3381..3400)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3541..3563
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3557..3748)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4557..5639)
                     /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     5601..5620
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5640..5717)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5923..5942
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6026..6045
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6026..6044
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6045..6069
                     /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             6084..6106
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 atggctagca tgactggtgg acagcaaatg ggtcgcggat ccatgttgag acgtgcggtg
       61 aatatatcca ttgccagagg gcgcatggcg ttgatgagct acgcgacgct gcctgacctt
      121 ttaaaaccaa gtggcgcccc ggccgagttg cccaagttgg ggtttaactg gaaggatgga
      181 tgtgcccctg tcttcagccc gcgccagatg gaattgcact acaccaaaca ccacaaggcg
      241 tatgtggaca aattaaatgc gctggcaggg acaacatacg atgggaagag cattgaggag
      301 attatccttg cggtcgcaaa tgacgccgag aagaagggtc ttttcaacca agccgcacag
      361 cattttaacc acacctttta cttccgttgc atcaccccta atgggaaggc gatgccaaag
      421 tcccttgagt ctgctgttac ggcccagttt ggttcggtgg aacagtttaa agatgccttt
      481 gtgcaggcag gggtgaataa ctttggctcg ggttggacgt ggctgtgcgt ggacccgtcg
      541 aataagaatc agctcgttat tgacaatacg agcaacgccg gatgtccctt aacgaaggga
      601 ctgcgccccg tcctcgcagt ggatgtgtgg gagcatgcgt actacaagga ctttgaaaac
      661 cgccggccgg attacttgaa ggagatctgg tcggtcatcg actgggagtt tgttgcaaag
      721 atgcatgtgc aggcaataaa agcggccgca ctcgagcacc accaccacca ccactgagat
      781 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
      841 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
      901 actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
      961 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1021 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1081 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     1141 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     1201 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     1261 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     1321 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     1381 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1441 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1501 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1561 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1621 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1681 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1741 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1801 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1861 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1921 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1981 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2041 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2101 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2161 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2221 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2281 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2341 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2401 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2461 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2521 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2581 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2641 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2701 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2761 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2821 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2881 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2941 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3001 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3061 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3121 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3181 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3241 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3301 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3361 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     3421 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     3481 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3541 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     3601 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     3661 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     3721 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     3781 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     3841 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     3901 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     3961 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     4021 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     4081 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     4141 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     4201 tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     4261 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     4321 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     4381 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     4441 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     4501 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     4561 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4621 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     4681 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     4741 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     4801 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
     4861 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     4921 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     4981 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     5041 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     5101 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     5161 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     5221 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     5281 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     5341 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     5401 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     5461 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     5521 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     5581 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     5641 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     5701 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     5761 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     5821 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     5881 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     5941 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     6001 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
     6061 aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata cat
//
