LOCUS       sequence-493092-        7632 bp    DNA     circular SYN 18-NOV-2025
DEFINITION  Expresses full-length human LGP2 in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7632)
  AUTHORS   Lee KY, Craig C, Patel SS
  TITLE     Unraveling blunt-end RNA binding and ATPase-driven translocation 
            activities of the RIG-I family helicase LGP2.
  JOURNAL   Nucleic Acids Res. 2024 Jan 11;52(1):355-369. doi: 
            10.1093/nar/gkad1106.
  PUBMED    38015453
REFERENCE   2  (bases 1 to 7632)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 18, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2024-01-11- 11"; volume: "52"; issue: 
            "1"; pages: "355-369"
COMMENT     Sequence Label: pET28_WT-LGP2 (codon optimized)
FEATURES             Location/Qualifiers
     source          1..7632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1137..1430)
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             complement(1443..1460)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1470..1487)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1506..1528
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1543..1567
                     /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)."
     primer_bind     complement(1567..1586)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1568..1586)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1670..1689)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1895..1972
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1973..3055
                     /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     complement(1992..2011)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3864..4055
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4049..4071)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4157..4296
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4212..4231
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4311..4328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4482..5070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4562..4581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5181..5200)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5192..6007
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             5192..6007
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5265..5284)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6099..6554)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6236..6257)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6448..6467
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(6591..6638)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     6634..6652
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(6705..6722)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tgacgggtac gggtaaaaat gatgccacgc gggctgttgc tgctgctgaa ttgacgctgc
       61 aggatctttt ccagcatctc cagtttcggg ttctccggac catgggtcgc caggtgcgcc
      121 agttcgttct tacgatcgtc aaacagcgcc agcagacgac gctccgcgca cagaatctgg
      181 gttttggtaa cgtgttcacg gtgatagaaa tcttgcagcg ccgccagcgc atccaccgca
      241 cgaacggtat cgtggatcag cagcgcgtcg ttataacgac gcaggtgcag cgcgtacaca
      301 cgttgctcct gcagacccgc cagcgccgcc gcctcgctca gcttcacaac ttgctgttca
      361 tacatctggg tgccaaattt acggctcagt tccggcatct ccaggtgatc gtgaatttgg
      421 tccatcagtt tcttcagcag atcaccgaac gggtcctggc tacgacggtg gcacaggttg
      481 tattgcttgc acggttgctg gctgtgttct tgcagctgcg ggcagcagtt ctgcgggctc
      541 ataatgcacc aggtgtccag gttcgcgcac agttgcagaa cgtggttgat cgcgccatcc
      601 agtttgctcg cgccaccggt acccgggctc gcggtcagac ccagcacctg cggcagcggc
      661 tgcgcacgtt gcagcttcag ctccagatat tggctcataa tcacgttgta aacggtgtct
      721 ttgtgggtgt ggtggcactc atccacaacg atcaggctga acacggtcag ttcaacgtgt
      781 tcctcttcct ccgggctggt cagcgccatc tgcagcagtt ccgcggtgca gatcagcagg
      841 tcatggcaac gcgccagatg accaaaaccc gcacgcggac ccatgtcacc gctcagggtg
      901 gtaacggtcc aacggccatc cagcatacga cggaattcct cgccgtgctg ggtcaccagg
      961 tgaacacggt tcaccagcac aaccactttc gcaccatcca cggtttccag gtgacgcttc
     1021 gcaacatacg ccgccgcacg ggttttgccc gcaccggtcg gcagccagat gatgatgttt
     1081 ttaccctcca gcgccggcat aataacttcc cattgatagc tacgcagttc catggatcca
     1141 ccaatctgtt ctctgtgagc ctcaataata tcgttatcct ccatgtccaa atcttcaggg
     1201 gtctgatcag cttgaattct aataccgtcg tacaagaatc ttaaggagtc catttcctta
     1261 ccctgtcttt tagcgaacgc ttccatcagc cttcttaaag gagtggtctt tttgatcttg
     1321 aagaagatct ctgaagatcc atcggacacc tttaaattga tgtgagtctc aggcttgact
     1381 tctggcttga cctctggctt agcttcttga ttgacttctg agtccgacat gctagccata
     1441 tggctgccgc gcggcaccag gccgctgctg tgatgatgat gatgatggct gctgcccatg
     1501 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
     1561 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
     1621 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     1681 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     1741 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
     1801 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
     1861 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
     1921 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     1981 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     2041 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     2101 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     2161 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     2221 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     2281 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     2341 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     2401 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     2461 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     2521 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     2581 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     2641 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     2701 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     2761 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     2821 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     2881 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     2941 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     3001 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     3061 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
     3121 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     3181 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     3241 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
     3301 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     3361 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
     3421 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
     3481 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
     3541 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
     3601 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
     3661 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
     3721 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
     3781 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
     3841 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
     3901 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
     3961 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
     4021 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     4081 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     4141 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     4201 gcttaactat gcggcatcag agcagattgt actgagagtg caccattgcg gtgtgaaata
     4261 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     4321 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4381 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4441 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4501 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4561 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4621 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4681 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4741 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4801 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4861 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4921 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4981 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5041 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5101 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc
     5161 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
     5221 ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     5281 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     5341 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     5401 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     5461 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
     5521 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     5581 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     5641 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     5701 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     5761 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     5821 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     5881 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     5941 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     6001 tttctaagaa ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6061 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     6121 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     6181 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     6241 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     6301 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     6361 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     6421 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     6481 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     6541 gctacagggc gcgtcccatt cgccaatccg gatatagttc ctcctttcag caaaaaaccc
     6601 ctcaagaccc gtttagaggc cccaaggggt tatgctagtt attgctcagc ggtggcagca
     6661 gccaactcag cttcctttcg ggctttgtta gcagccggat ctcagtggtg gtggtggtgg
     6721 tgctcgagtt agtccaggct caggtcgctc aggttctccg cgcagtgttg caggaaatca
     6781 aaatccggca cgctaaacgg aacacggctc catttcttcg cctgaatacg accctgcggg
     6841 gtttccagca gcatgctacg cactttcaga accggcagct taacgctttt atagatcatc
     6901 tgcaggcccc aaacttcacc gcagttacgg cagctaatca cgccacccgg cttccagtct
     6961 ttaaaaacct tgttgataac caccggatca cggctaacgt tatagtagtt gctgaagttc
     7021 gggttaacgt tcacgtggtg ggtgccctcc accttacgca ggtcgctgcc atgaccaacc
     7081 gccaccatgc agttgatgca cagcagctgc acgtgctcaa ccgggaattg ctgacgttgg
     7141 ttttcacgtt gcgccgcctg cgccgcacgc ttggtcagcg ccgcttgctg caggtcacga
     7201 atcttcgcct ggtactccgc ttgatccatt ttctgcaccg ccgcaaccgc ctgttccatc
     7261 agggtttcca gcgcctcgtt gatcagctca cgcttcagtt cacggctacc ctcggtcgcc
     7321 acaaacgcat aaacgctctg gtccgcccgc gcacgaccac gcgcttgcac catgctaatc
     7381 tcgttggtca gcaggccgta acgaaccaca acgttgcagt gcgggatatc cagaccttcc
     7441 tccgccacgc tggtcgcaac cagcaggttc agggtgccgt cttggaattt ctgaataact
     7501 tcttgctgat cacgctgggt catgtgggtg ctttggctgc tgttgcccgc accaatcagc
     7561 agctgcgcac ggatgtccac ggtttgcaga ccttgctgtt gctgcagcca cagcagcagg
     7621 ctgtgcgcgc tc
//
