LOCUS       sequence-246329-        7985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the Helicobacter hepaticus specific TCRa/b HH5-5 in T cell 
            hybridoma cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7985)
  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 7985)
  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-HH5-5
FEATURES             Location/Qualifiers
     source          1..7985
                     /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"
     CDS             2243..2299
                     /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    3234..3801
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3251..3803
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3395..3412)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3622..3641
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3825..4313
                     /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             4380..4894
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5056..5072
                     /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(5080..5086,5087..5104,5105..5110))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5125..5146
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5263..5280)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5434..6022)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5514..5533)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6193..6984,6985..7053))
                     /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     6816..6835
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7054..7158)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7226..7244
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7282..7304)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7404..7423
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7617..7639
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7767..7786
                     /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 tgctcctggt
     1441 tctcatctcg ttcctcggga tacatttctt cctggatgtc caaacacaga cagtttccca
     1501 gtctgatgcc catgtcactg tcttcgaagg agactcggtg gagctgagat gcaactattc
     1561 ctatggtgga tccatttacc tctcctggta catccagcac catggccatg gcctccagtt
     1621 tctcctcaag tactattcgg gaaacccagt ggttcaagga gtgaacggct tcgaggctga
     1681 gttcagcaag agcgactctt ccttccacct tcggaaagcc tctgtgcact ggagcgactc
     1741 ggctgtgtac ttctgtgctg tgatctggtc taattacaac gtgctttact tcggatctgg
     1801 caccaaactc actgtagagc caaacatcca gaacccagaa cctgctgtgt accagttaaa
     1861 agatcctcgg tctcaggaca gcaccctctg cctgttcacc gactttgact cccaaatcaa
     1921 tgtgccgaaa accatggaat ctggaacgtt catcactgac aaaactgtgc tggacatgaa
     1981 agctatggat tccaagagca atggggccat tgcctggagc aaccagacaa gcttcacctg
     2041 ccaagatatc ttcaaagaga ccaacgccac ctaccccagt tcagacgttc cctgtgatgc
     2101 cacgttgact gagaaaagct ttgaaacaga tatgaaccta aactttcaaa acctgtcagt
     2161 tatgggactc cgaatcctcc tgctgaaagt agccggattt aacctgctca tgacgctgag
     2221 gctgtggtcc agtggctccg gagccacgaa cttctctctg ttaaagcaag caggagacgt
     2281 ggaagaaaac cccggtccca tgaacaagtg ggttttctgc tgggtaaccc tttgtctcct
     2341 tactgtagag accacacatg gtgatggtgg catcattact cagacaccca aattcctgat
     2401 tggtcaggaa gggcaaaaac tgaccttgaa atgtcaacag aatttcaatc atgatacaat
     2461 gtactggtac cgacaggatt cagggaaagg attgagactg atctactatt caataactga
     2521 aaacgatctt caaaaaggcg atctatctga aggctatgat gcgtctcgcg agaagaagtc
     2581 atctttttct ctcactgtga catctgccca gaagaacgag atggccgttt ttctctgtgc
     2641 cagcagtaga accggggggt acaccgggca gctctacttt ggtgaaggct caaagctgac
     2701 agtgctggaa gatctgagaa atgtgactcc acccaaggtc tccttgtttg agccatcaaa
     2761 agcagagatt gcaaacaaac aaaaggctac cctcgtgtgc ttggccaggg gcttcttccc
     2821 tgaccacgtg gagctgagct ggtgggtgaa tggcaaggag gtccacagtg gggtcagcac
     2881 ggaccctcag gcctacaagg agagcaatta tagctactgc ctgagcagcc gcctgagggt
     2941 ctctgctacc ttctggcaca atcctcgaaa ccacttccgc tgccaagtgc agttccatgg
     3001 gctttcagag gaggacaagt ggccagaggg ctcacccaaa cctgtcacac agaacatcag
     3061 tgcagaggcc tggggccgag cagactgtgg aatcacttca gcatcctatc atcagggggt
     3121 tctgtctgca accatcctct atgagatcct actggggaag gccaccctat atgctgtgct
     3181 ggtcagtggc ctggtgctga tggccatggt caagaaaaaa aattcctgag aattccgccc
     3241 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     3301 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     3361 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     3421 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     3481 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     3541 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3601 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     3661 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     3721 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3781 ctttgaaaaa cacgatgata atatggccac aaccatgggc caccatgaac ccagccatca
     3841 gcgtcgctct cctgctctca gtcttgcagg tgtcccgagg gcagaaggtg accagcctga
     3901 cagcctgcct ggtgaaccaa aaccttcgcc tggactgccg ccatgagaat aacaccaagg
     3961 ataactccat ccagcatgag ttcagcctga cccgagagaa gaggaagcac gtgctctcag
     4021 gcacccttgg gatacccgag cacacgtacc gctcccgcgt caccctctcc aaccagccct
     4081 atatcaaggt ccttacccta gccaacttca ccaccaagga tgagggcgac tacttttgtg
     4141 agcttcgcgt ctcgggcgcg aatcccatga gctccaataa aagtatcagt gtgtatagag
     4201 acaagctggt caagtgtggc ggcataagcc tgctggttca gaacacatcc tggatgctgc
     4261 tgctgctgct ttccctctcc ctcctccaag ccctggactt catttctctg tgagtcgacc
     4321 tgcagccaag cttatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     4381 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     4441 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
     4501 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     4561 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
     4621 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     4681 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     4741 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata
     4801 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     4861 agtgattgac tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga
     4921 acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt
     4981 tgaatccaca tactccaata ctcctgaaat agttcattat ggacagcgca gaaagagctg
     5041 gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5101 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5161 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5221 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5281 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5341 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5401 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5461 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5521 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5581 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5641 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5701 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5761 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5821 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5881 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5941 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6001 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6061 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6121 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6181 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6241 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6301 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6361 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6421 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6481 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6541 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6601 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6661 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6721 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6781 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6841 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6901 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6961 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7021 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7081 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7141 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     7201 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     7261 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     7321 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     7381 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     7441 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     7501 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     7561 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     7621 gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac
     7681 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     7741 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     7801 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt
     7861 gttaaagaca ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg
     7921 aagcctatag agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag
     7981 ggggg
//
