LOCUS       sequence-246328-        8009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the Helicobacter hepaticus specific TCRa/b HH5-4 in T cell 
            hybridoma cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8009)
  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 8009)
  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: RV-mCD4-HH5-4
FEATURES             Location/Qualifiers
     source          1..8009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1424..1433
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2270..2326
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    3258..3825
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3275..3827
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3419..3436)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3646..3665
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3849..4337
                     /codon_start=1
                     /product="mouse GPI-anchored cell surface glycoprotein of 
                     the immunoglobulin superfamily"
                     /label=Thy1.1
                     /note="This allele of Thy1 is also known as CD90.1."
                     /translation="MNPAISVALLLSVLQVSRGQKVTSLTACLVNQNLRLDCRHENNTK
                     DNSIQHEFSLTREKRKHVLSGTLGIPEHTYRSRVTLSNQPYIKVLTLANFTTKDEGDYF
                     CELRVSGANPMSSNKSISVYRDKLVKCGGISLLVQNTSWMLLLLLSLSLLQALDFISL"
     LTR             4404..4918
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5080..5096
                     /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)."
     promoter        complement(join(5104..5110,5111..5128,5129..5134))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5149..5170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5287..5304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5458..6046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5538..5557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6217..7008,7009..7077))
                     /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     6840..6859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7078..7182)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7250..7268
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7306..7328)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7428..7447
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7641..7663
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7791..7810
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatctctc gaggccacca tggacaagat
     1441 tctgacagca tcatttttac tcctaggcct tcacctagct ggggtgaatg gccagcagaa
     1501 ggagaaacat gaccagcagc aggtgagaca aagtccccaa tctctgacag tctgggaagg
     1561 aggaaccaca gttctgacct gcagttatga ggacagcact tttaactact tcccatggta
     1621 ccaacagttc cctggggaag gccctgcact tctgatatcc atactttcag tgtccgataa
     1681 aaaggaagat ggacgattca caaccttctt caataaaagg gagaaaaagc tctccttgca
     1741 catcatagac tctcagcctg gagactcagc cacctacttc tgtgcagcaa gtccggctgg
     1801 aggctataaa gtggtctttg gaagtgggac tcgattgctg gtaagccctg acatccagaa
     1861 cccagaacct gctgtgtacc agttaaaaga tcctcggtct caggacagca ccctctgcct
     1921 gttcaccgac tttgactccc aaatcaatgt gccgaaaacc atggaatctg gaacgttcat
     1981 cactgacaaa actgtgctgg acatgaaagc tatggattcc aagagcaatg gggccattgc
     2041 ctggagcaac cagacaagct tcacctgcca agatatcttc aaagagacca acgccaccta
     2101 ccccagttca gacgttccct gtgatgccac gttgactgag aaaagctttg aaacagatat
     2161 gaacctaaac tttcaaaacc tgtcagttat gggactccga atcctcctgc tgaaagtagc
     2221 cggatttaac ctgctcatga cgctgaggct gtggtccagt ggctccggag ccacgaactt
     2281 ctctctgtta aagcaagcag gagacgtgga agaaaacccc ggtcccatgt ggcagttttg
     2341 cattctgtgc ctctgtgtac tcatggcttc tgtggctaca gaccccacag tgactttgct
     2401 ggagcaaaac ccaaggtggc gtctggtacc acgtggtcaa gctgtgaacc tacgctgcat
     2461 cttgaaaaat tcccagtatc cctggatgag ctggtatcag caggatctcc aaaagcaact
     2521 acagtggctg ttcactctgc ggagtcctgg ggacaaagag gtcaaatctc ttcccggtgc
     2581 tgattacctg gccacacggg tcactgatac ggagctgagg ctgcaagtgg ccaacatgag
     2641 ccagggcaga accttgtact gcacctgcag ttcccgacag gggtcaaaca cagaagtctt
     2701 ctttggtaaa ggaaccagac tcacagttgt agaagatctg agaaatgtga ctccacccaa
     2761 ggtctccttg tttgagccat caaaagcaga gattgcaaac aaacaaaagg ctaccctcgt
     2821 gtgcttggcc aggggcttct tccctgacca cgtggagctg agctggtggg tgaatggcaa
     2881 ggaggtccac agtggggtca gcacggaccc tcaggcctac aaggagagca attatagcta
     2941 ctgcctgagc agccgcctga gggtctctgc taccttctgg cacaatcctc gaaaccactt
     3001 ccgctgccaa gtgcagttcc atgggctttc agaggaggac aagtggccag agggctcacc
     3061 caaacctgtc acacagaaca tcagtgcaga ggcctggggc cgagcagact gtggaatcac
     3121 ttcagcatcc tatcatcagg gggttctgtc tgcaaccatc ctctatgaga tcctactggg
     3181 gaaggccacc ctatatgctg tgctggtcag tggcctggtg ctgatggcca tggtcaagaa
     3241 aaaaaattcc tgagaattcc gccccccccc cctaacgtta ctggccgaag ccgcttggaa
     3301 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     3361 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     3421 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     3481 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     3541 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     3601 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     3661 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     3721 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     3781 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     3841 gggccaccat gaacccagcc atcagcgtcg ctctcctgct ctcagtcttg caggtgtccc
     3901 gagggcagaa ggtgaccagc ctgacagcct gcctggtgaa ccaaaacctt cgcctggact
     3961 gccgccatga gaataacacc aaggataact ccatccagca tgagttcagc ctgacccgag
     4021 agaagaggaa gcacgtgctc tcaggcaccc ttgggatacc cgagcacacg taccgctccc
     4081 gcgtcaccct ctccaaccag ccctatatca aggtccttac cctagccaac ttcaccacca
     4141 aggatgaggg cgactacttt tgtgagcttc gcgtctcggg cgcgaatccc atgagctcca
     4201 ataaaagtat cagtgtgtat agagacaagc tggtcaagtg tggcggcata agcctgctgg
     4261 ttcagaacac atcctggatg ctgctgctgc tgctttccct ctccctcctc caagccctgg
     4321 acttcatttc tctgtgagtc gacctgcagc caagcttatc gataaaataa aagattttat
     4381 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     4441 aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc
     4501 aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt
     4561 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag
     4621 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     4681 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     4741 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc
     4801 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     4861 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg
     4921 gtaacagttt cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa
     4981 tcctgactca attagccact gttttgaatc cacatactcc aatactcctg aaatagttca
     5041 ttatggacag cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc
     5101 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5161 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5221 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5281 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5341 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5401 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5461 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5521 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5581 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5641 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5701 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5761 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5821 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5881 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5941 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6001 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6061 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6121 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6181 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6241 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6301 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6361 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6421 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6481 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6541 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6601 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6661 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6721 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6781 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6841 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6901 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6961 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7021 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7081 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7141 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7201 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     7261 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     7321 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     7381 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     7441 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     7501 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     7561 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     7621 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt
     7681 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     7741 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     7801 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     7861 gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt
     7921 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
     7981 attttattta gtctccagaa aaagggggg
//
