LOCUS       sequence-250176-        6094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression of recombinant H. hepaticus antigen HH_1713.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6094)
  AUTHORS   Xu M, Pokrovskii M, Ding Y, Yi R, Au C, Harrison OJ, Galan C, 
            Belkaid Y, Bonneau R, Littman DR
  TITLE     c-MAF-dependent regulatory T cells mediate immunological tolerance 
            to a gut pathobiont.
  JOURNAL   Nature. 2018 Feb 15;554(7692):373-377. doi: 10.1038/nature25500. 
            Epub 2018 Feb 7.
  PUBMED    29414937
REFERENCE   2  (bases 1 to 6094)
  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/nature25500"; journalName: "Nature"; date: "2018-02-15- 
            15"; volume: "554"; issue: "7692"; pages: "373-377"
COMMENT     Sequence Label: pGEX6p1-HH1713
FEATURES             Location/Qualifiers
     source          1..6094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(197..202,203..218,219..225)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    233..249
                     /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)."
     CDS             272..925
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     883..905
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             932..955
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2158..2180)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2325..2343)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2411..2515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2516..2584,2585..3376)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2734..2753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3547..4135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4036..4055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4289..4306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4379..4456
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4457..5539
                     /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(4476..4495)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5588..5593,5594..5611,5612..5618)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5626..5642
                     /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     5650..5666
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5650..5666
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5662..6094,1..95)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5682..5699)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5682..5698)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5691..5713)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 atggcgttgg aattagctta tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc
       61 catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca
      121 aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat
      181 attctgaaat gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc
      241 ggataacaat ttcacacagg aaacagtatt catgtcccct atactaggtt attggaaaat
      301 taagggcctt gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga
      361 gcatttgtat gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt
      421 ggagtttccc aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc
      481 catcatacgt tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc
      541 agagatttca atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc
      601 atatagtaaa gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct
      661 gaaaatgttc gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca
      721 tcctgacttc atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct
      781 ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga
      841 taagtacttg aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt
      901 tggtggtggc gaccatcctc caaaatcgga tctggaagtt ctgttccagg ggcccctggg
      961 atccatgaaa aaaattgttg taagtagtgc tttatcagct cttcttgttt taccagcagc
     1021 agcttttgaa gtatataatg ctgatgatac taaagttgat ttatatggtt ctattcgtgg
     1081 ttatatgggt tatggtgaaa ctggtaatgg tggagcaaat gcaggatacc taattggggt
     1141 gcaaaataac tcacaatttg gtgtgaaggt aaaatctggt aaatttagtg caaaagtaga
     1201 acttggtgca aaagaaacag gtataagcgg tgatggcaca ccttatcgcg aatattgggg
     1261 agcttatgat acaaatatag gacaaatcct ttttggtaaa gcaatgtctc caacgattga
     1321 taatgctttc acaagcaact ggttaaataa cgataatggc tatcaagggg ttggtggtgt
     1381 cgctacaggt aatcgtcaaa tccaacttca atataatgta gcaggattga gtcttgcact
     1441 tattgaagta actgctggta atggtagggg agatttaaat caagaatccc ctcgtattgc
     1501 tgcagcttat acaatcaaag gtgcgaatgg acagccatta ttaaaaatcg cagggacata
     1561 taaatacttt aatgggctta ataatctaac tactcccagc ccattggctg gagctaatgc
     1621 gtggcacgta tgggctgggt taaaacctac ttttggtaac gcatatatct ctgtgctcgc
     1681 acattatggt aaaaatggac atctttatgg tgagcaaaac actacatata atggaggtgg
     1741 ctatttactt aaccctcttg acactgatat tgggcttaat gcaataagaa caggtgggct
     1801 tgtagagttt ggcacaaaat taagcagtga tttatctctt gcaattggtg caggatatca
     1861 agcaacattt aatggtgatg gtcaagcaac aacaggtgga acatcagtta gtagttatgg
     1921 aaaactacac ggatattctg cgtttatcaa tctcccttat aaagtcaatg caaacttcac
     1981 ttttgctcct caaatagctt attatgggct ttcaggtagt gaaagagtag caaatgcaca
     2041 atataaacga acaggtgtgg ttgctgctac acgatttaaa tgggatttct aactcgagcg
     2101 gccgcatcgt gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     2161 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     2221 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     2281 gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct
     2341 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     2401 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2461 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2521 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2581 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2641 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2701 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     2761 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2821 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2881 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2941 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3001 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     3061 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3121 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3181 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3241 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3301 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3361 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3421 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3481 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3541 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3601 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3661 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3721 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3781 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3841 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3901 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3961 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4021 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4081 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4141 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4201 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4261 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4321 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga
     4381 caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     4441 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     4501 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     4561 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
     4621 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     4681 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     4741 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     4801 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     4861 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     4921 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
     4981 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     5041 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
     5101 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
     5161 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
     5221 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
     5281 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     5341 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     5401 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc acgctagcgc ccaatacgca
     5461 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     5521 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
     5581 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     5641 aatttcacac aggaaacagc tatgaccatg attacggatt cactggccgt cgttttacaa
     5701 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     5761 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     5821 agcctgaatg gcgaatggcg ctttgcctgg tttccggcac cagaagcggt gccggaaagc
     5881 tggctggagt gcgatcttcc tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg
     5941 cacggttacg atgcgcccat ctacaccaac gtgacctatc ccattacggt caatccgccg
     6001 tttgttccca cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc
     6061 tggctacagg aaggccagac gcgaattatt tttg
//
