LOCUS       sequence-246355-        7967 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the Helicobacter hepaticus specific TCRa/b HH7-9 in T cell 
            hybridoma cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7967)
  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 7967)
  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-HH7-9
FEATURES             Location/Qualifiers
     source          1..7967
                     /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    3216..3783
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3233..3785
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3377..3394)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3604..3623
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3807..4295
                     /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             4362..4876
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5038..5054
                     /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(5062..5068,5069..5086,5087..5092))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5107..5128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5245..5262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5416..6004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5496..5515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6175..6966,6967..7035))
                     /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     6798..6817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7036..7140)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7208..7226
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7264..7286)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7386..7405
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7599..7621
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7749..7768
                     /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 tgagagggag cagtggcaac aagctcatct ttggaattgg
     1801 gactctgctt tctgtcaagc 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 tgctgtactc tctccttgcc tttctcctgg gcatgttctt
     2341 gggtgttagt gctcagacta tccatcaatg gccagttgcc gagatcaagg ctgtgggcag
     2401 cccactgtct ctggggtgta ccataaaggg gaaatcaagc cctaacctct actggtactg
     2461 gcaggccaca ggaggcaccc tccagcaact cttctactct attactgttg gccaggtaga
     2521 gtcggtggtg caactgaacc tctcagcttc caggccgaag gacgaccaat tcatcctaag
     2581 cacggagaag ctgcttctca gccactctgg cttctacctc tgtgcctgga gtgccgacag
     2641 ggccaacaca gaagtcttct ttggtaaagg aaccagactc acagttgtag aagatctgag
     2701 aaatgtgact ccacccaagg tctccttgtt tgagccatca aaagcagaga ttgcaaacaa
     2761 acaaaaggct accctcgtgt gcttggccag gggcttcttc cctgaccacg tggagctgag
     2821 ctggtgggtg aatggcaagg aggtccacag tggggtcagc acggaccctc aggcctacaa
     2881 ggagagcaat tatagctact gcctgagcag ccgcctgagg gtctctgcta ccttctggca
     2941 caatcctcga aaccacttcc gctgccaagt gcagttccat gggctttcag aggaggacaa
     3001 gtggccagag ggctcaccca aacctgtcac acagaacatc agtgcagagg cctggggccg
     3061 agcagactgt ggaatcactt cagcatccta tcatcagggg gttctgtctg caaccatcct
     3121 ctatgagatc ctactgggga aggccaccct atatgctgtg ctggtcagtg gcctggtgct
     3181 gatggccatg gtcaagaaaa aaaattcctg agaattccgc cccccccccc taacgttact
     3241 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     3301 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     3361 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     3421 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     3481 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     3541 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     3601 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     3661 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
     3721 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     3781 taatatggcc acaaccatgg gccaccatga acccagccat cagcgtcgct ctcctgctct
     3841 cagtcttgca ggtgtcccga gggcagaagg tgaccagcct gacagcctgc ctggtgaacc
     3901 aaaaccttcg cctggactgc cgccatgaga ataacaccaa ggataactcc atccagcatg
     3961 agttcagcct gacccgagag aagaggaagc acgtgctctc aggcaccctt gggatacccg
     4021 agcacacgta ccgctcccgc gtcaccctct ccaaccagcc ctatatcaag gtccttaccc
     4081 tagccaactt caccaccaag gatgagggcg actacttttg tgagcttcgc gtctcgggcg
     4141 cgaatcccat gagctccaat aaaagtatca gtgtgtatag agacaagctg gtcaagtgtg
     4201 gcggcataag cctgctggtt cagaacacat cctggatgct gctgctgctg ctttccctct
     4261 ccctcctcca agccctggac ttcatttctc tgtgagtcga cctgcagcca agcttatcga
     4321 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     4381 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag
     4441 aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg
     4501 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     4561 cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     4621 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     4681 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt
     4741 cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
     4801 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     4861 agcgggggtc tttcatgggt aacagtttct tgaagttgga gaacaacatt ctgagggtag
     4921 gagtcgaata ttaagtaatc ctgactcaat tagccactgt tttgaatcca catactccaa
     4981 tactcctgaa atagttcatt atggacagcg cagaaagagc tggggagaat tgtgaaattg
     5041 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5101 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5161 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5221 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5281 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5341 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5401 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5461 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5521 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5581 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5641 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5701 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5761 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5821 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5881 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5941 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6001 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6061 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6121 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6181 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6241 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6301 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6361 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6421 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6481 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6541 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6601 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6661 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6721 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6781 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6841 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6901 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6961 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7021 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7081 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7141 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     7201 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     7261 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     7321 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     7381 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     7441 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     7501 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     7561 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     7621 gacggcgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     7681 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     7741 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     7801 ccggccacga tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga caggatatca
     7861 gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag
     7921 ccatagataa aataaaagat tttatttagt ctccagaaaa agggggg
//
