LOCUS       sequence-438971-        6805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of STING CTT in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6805)
  AUTHORS   Landau LM, Chaudhary N, Tien YC, Rogozinska M, Joshi S, Yao C, 
            Crowley J, Hullahalli K, Campbell IW, Waldor MK, Haigis M, Kagan JC
  TITLE     pLxIS-containing domains are biochemically flexible regulators of 
            interferons and metabolism.
  JOURNAL   Mol Cell. 2024 Jul 11;84(13):2436-2454.e10. doi: 
            10.1016/j.molcel.2024.05.030. Epub 2024 Jun 25.
  PUBMED    38925114
REFERENCE   2  (bases 1 to 6805)
  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: 
            "10.1016/j.molcel.2024.05.030"; journalName: "Mol Cell"; date: 
            "2024-07-11- 11"; volume: "84"; issue: "13"; pages: "2436-2454.e10"
COMMENT     Sequence Label: pET28a-FKBP-STING(pLxIS)
FEATURES             Location/Qualifiers
     source          1..6805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      7..16
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1499..1516
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1569..1587)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1583..1630
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1667..2122
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1754..1773)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1964..1985
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2214..3029)
                     /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             complement(2214..3029)
                     /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     2937..2956
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3021..3040
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3151..3739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3640..3659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3893..3910
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3925..4064
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3990..4009)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4150..4172
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4166..4357)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5166..6248)
                     /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     6210..6229
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6249..6326)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6532..6551
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6635..6654
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6635..6653
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6654..6678
                     /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             6693..6715
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6734..6751
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6761..6778
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             6785..6805
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 ggatccgcca ccatgggcgt tcaagtagaa acaatatctc caggagacgg gcgtacattt
       61 cctaagcgtg gccaaacttg cgtagtccat tataccggca tgctggagga cggtaagaag
      121 gtcgatagct ctagagaccg taacaaacct ttcaaattta tgctggggaa acaggaggtt
      181 atccgtggtt gggaggaagg cgtggcccaa atgtccgttg ggcagcgtgc taagttaacc
      241 atctcgccag attacgcata tggggctaca ggccatcctg ggatcattcc cccgcatgcg
      301 acacttgtgt ttgatgtaga gctgttgaag ttggagacgc ggggagttca agtcgaaacg
      361 atttcaccag gtgatggacg cacctttcct aaacggggac aaacgtgcgt cgtgcattac
      421 accggtatgc tggaagacgg caagaaagtc gattcaagcc gcgatagaaa taaacccttc
      481 aaattcatgc ttggaaaaca ggaagtaata cgcggatggg aagagggtgt ggcgcaaatg
      541 tctgtcgggc aaagagctaa actgacaata agtcctgatt acgcgtacgg tgcgactggg
      601 caccctggta tcattccgcc gcacgcgaca cttgtcttcg acgtcgagtt acttaagctt
      661 gagacgcggg gggtgcaggt agagactatt tcgcccggtg acggaagaac tttcccaaag
      721 cggggacaga cctgtgttgt ccactacact gggatgttgg aggatggtaa gaaagttgac
      781 tcaagtcgtg atcgcaacaa accctttaag tttatgcttg ggaagcagga agtcatcaga
      841 ggttgggaag aaggcgtagc tcagatgtca gtgggacaac gggccaaatt aaccatcagt
      901 ccagactacg cctacggagc caccggtcac ccagggatca taccacctca cgcaacctta
      961 gtgttcgatg tggaactttt aaagttagag acgcgcggtg tacaagtgga aacaatcagt
     1021 ccaggggatg ggagaacatt ccccaaaaga ggccagacgt gtgtcgtgca ctatacgggg
     1081 atgttagaag atgggaaaaa ggtcgattcg tcacgcgacc gcaacaaacc gtttaaattc
     1141 atgttgggaa agcaggaggt gattcgcggc tgggaggaag gtgtggcaca gatgagcgtg
     1201 ggccagagag caaaactgac tatctctcct gattatgcct acggcgcgac gggccaccct
     1261 ggcataattc caccccacgc taccctggta ttcgatgtcg agcttctgaa gcttgagggt
     1321 gctggggctg gggctgttac aatgaatgcg cctatgacta gtgtggcccc gccgccttcg
     1381 gttttatcac aggagccgcg gctgttaatt agcggtatgg atcaaccgct gcctcttcgg
     1441 accgatttga tctgataaga attcgagctc cgtcgacaag cttgcggccg cactcgagca
     1501 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     1561 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     1621 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
     1681 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1741 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1801 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1861 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1921 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1981 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2041 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2101 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2161 tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa
     2221 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     2281 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     2341 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     2401 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     2461 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     2521 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     2581 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2641 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     2701 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     2761 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     2821 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     2881 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     2941 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag
     3001 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     3061 gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3121 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3181 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3241 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3301 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3361 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3421 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3481 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3541 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3601 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3661 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3721 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3781 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3841 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3901 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3961 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4021 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     4081 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4141 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4201 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     4261 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     4321 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     4381 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     4441 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     4501 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     4561 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     4621 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     4681 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     4741 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     4801 ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc
     4861 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     4921 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     4981 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     5041 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     5101 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     5161 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5221 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     5281 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     5341 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     5401 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     5461 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     5521 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     5581 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     5641 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     5701 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     5761 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     5821 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     5881 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     5941 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     6001 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     6061 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     6121 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     6181 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     6241 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     6301 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     6361 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     6421 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     6481 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     6541 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     6601 ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg
     6661 tgagcggata acaattcccc tctagaaata attttgttta actttaagaa ggagatatac
     6721 catgggcagc agccatcatc atcatcatca cagcagcggc ctggtgccgc gcggcagcca
     6781 tatggaaaac ctgtattttc agggc
//
