LOCUS       sequence-481078-        6781 bp    DNA     circular SYN 29-JUL-2025
DEFINITION  Expresses LPETG-tagged, constitutively active human IRF5 variant in 
            bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6781)
  AUTHORS   Eweje F, Ibrahim V, Shajii A, Walsh ML, Ahmad K, Alrefai A, Miyasato
            D, Davis JR, Ham H, Li K, Roehrl M, Haller CA, Liu DR, Chen J, 
            Chaikof EL
  TITLE     Self-assembling protein nanoparticles for cytosolic delivery of 
            nucleic acids and proteins.
  JOURNAL   Nat Biotechnol. 2025 May 15. doi: 10.1038/s41587-025-02664-2.
  PUBMED    40374955
REFERENCE   2  (bases 1 to 6781)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 29, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2025 May 15. doi: 10.1038/s41587-025-02664-2."
COMMENT     Sequence Label: pET-hIRF5-V5(4D)-LPETG
FEATURES             Location/Qualifiers
     source          1..6781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1694..1711
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1730..1774
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1812..1830)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1826..1873
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1910..2365
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1997..2016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2207..2228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2480..3271,3272..3340))
                     /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     3103..3122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3341..3433)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3514..4102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4003..4022
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4256..4273
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4288..4427
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4353..4372)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4513..4535
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4529..4720)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5292..6374)
                     /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     6336..6355
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6375..6452)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6658..6677
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6765..6781,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(6765..6781,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgaatcaat caatacccgt agctccaaca ccgccacgtc gtgttcgcct
      121 gaagccgtgg ctggtggcac aagtcaatag ctgccagtac ccgggtttgc aatgggttaa
      181 cggggagaag aagttgttct gcattccgtg gcgtcacgcg acccgtcatg gtccgagcca
      241 ggacggcgat aacaccattt ttaaagcctg ggcaaaagag acaggtaaat ataccgaagg
      301 tgttgatgaa gcggacccgg cgaagtggaa agccaatctg cgttgcgctc tgaacaaaag
      361 ccgtgatttt aggctaattt atgatggtcc gcgcgacatg ccgccgcagc catacaaaat
      421 ctatgaagta tgtagcaacg gcccggcgcc gacggacagc cagccgcctg aagactacag
      481 ctttggagcg ggcgaggagg aggaggagga ggaggaactt caacgcatgc taccgtccct
      541 gtccctgacc gacgctgtgc aatccggccc acacatgacc ccgtactctc tgctgaaaga
      601 agacgtgaag tggccaccga ccttgcaacc gcctacttta agacctccga cccttcaacc
      661 accgaccctg caaccgccgg ttgttttggg cccaccggct ccggacccgt cgccgctggc
      721 gccgccgcct ggtaatccgg cgggcttccg cgaactgttg agcgaagttt tggagccggg
      781 tccgctgccg gcttcgttgc cgccggcggg tgagcagctg ctacccgacc tactcatctc
      841 cccgcacatg ctcccgctga ctgacctgga gattaagttc cagtatcgcg gtcgtccgcc
      901 gcgtgcactg acgatctcca accctcacgg ctgccgtttg ttctacagtc aattggaggc
      961 tacccaggag caggtcgagc tgttcggtcc gatttccctg gagcaggtac gttttccgag
     1021 cccggaggac atcccaagcg ataagcagcg tttctatacg aaccagctcc tggatgtgct
     1081 ggatcgcggc ctgatcctgc aactgcaagg tcaagatctg tacgcaattc gtctttgcca
     1141 gtgtaaagtg ttctggtcag gcccgtgtgc gtctgcgcac gatagctgcc cgaaccccat
     1201 ccagagagaa gttaagacca aattgttcag cttggagcac tttctgaatg aactgatcct
     1261 gttccagaaa ggtcagacca atactccgcc accgtttgaa atcttctttt gcttcggcga
     1321 ggaatggcct gaccgcaaac cgcgtgaaaa gaagctgatc accgttcaag tggtgccggt
     1381 tgccgcacgt ctgttactgg agatgttttc tggtgagctg gattgggatg cggatgatat
     1441 tcgcttacaa atcgataacc cggacctgaa ggaccgtatg gtggaacagt ttaaggaact
     1501 gcatcatatt tggcaaagcc agcagcgtct gcagccggtc gctcaagcgc ctccgggcgc
     1561 gggcttaggt gtgggtcagg gtccgtggcc gatgcatccg gcgggcatgc aggggggaag
     1621 tggtggatca agagtacgaa ggggcggtag cggtggctcc ctcccagaga ccggtgtcgt
     1681 agacggcgga tcccatcacc accaccatca ctaatgactc gagtctggta aagaaaccgc
     1741 tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt aattaaccta
     1801 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     1861 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
     1921 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1981 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2041 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2101 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2161 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2221 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2281 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2341 aacaaaatat taacgtttac aatttctggc ggcacgatgg catgagatta tcaaaaagga
     2401 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2461 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2521 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2581 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2641 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2701 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     2761 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2821 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     2881 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     2941 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3001 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3061 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3121 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3181 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3241 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3301 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatcat
     3361 gattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3421 aaaaataaac aaataggtca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3481 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3541 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3601 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3661 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3721 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3781 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3841 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3901 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3961 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4021 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4081 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4141 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4201 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4261 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     4321 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4381 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     4441 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4501 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4561 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     4621 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     4681 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     4741 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     4801 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     4861 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     4921 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     4981 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     5041 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     5101 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     5161 acaggagcac gatcatgcta gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     5221 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     5281 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5341 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     5401 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     5461 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     5521 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca
     5581 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     5641 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     5701 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     5761 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     5821 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     5881 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     5941 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     6001 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     6061 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     6121 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     6181 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     6241 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     6301 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     6361 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     6421 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga
     6481 ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag
     6541 gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
     6601 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
     6661 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
     6721 gcgtccggcg tagaggatcg agatcgatct cgatcccgcg aaattaatac gactcactat
     6781 a
//
