LOCUS       sequence-246391-        7976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the Helicobacter hepaticus specific TCRa/b HH7-11 in T cell 
            hybridoma cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7976)
  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 7976)
  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-HH7-11
FEATURES             Location/Qualifiers
     source          1..7976
                     /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             2246..2302
                     /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    3225..3792
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3242..3794
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3386..3403)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3613..3632
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3816..4304
                     /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             4371..4885
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5047..5063
                     /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(5071..5077,5078..5095,5096..5101))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5116..5137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5254..5271)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5425..6013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5505..5524)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6184..6975,6976..7044))
                     /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     6807..6826
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7045..7149)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7217..7235
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7273..7295)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7395..7414
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7608..7630
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7758..7777
                     /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 tgagccgcat gtctaattac aacgtgcttt acttcggatc
     1801 tggcaccaaa ctcactgtag agccaaacat ccagaaccca gaacctgctg tgtaccagtt
     1861 aaaagatcct cggtctcagg acagcaccct ctgcctgttc accgactttg actcccaaat
     1921 caatgtgccg aaaaccatgg aatctggaac gttcatcact gacaaaactg tgctggacat
     1981 gaaagctatg gattccaaga gcaatggggc cattgcctgg agcaaccaga caagcttcac
     2041 ctgccaagat atcttcaaag agaccaacgc cacctacccc agttcagacg ttccctgtga
     2101 tgccacgttg actgagaaaa gctttgaaac agatatgaac ctaaactttc aaaacctgtc
     2161 agttatggga ctccgaatcc tcctgctgaa agtagccgga tttaacctgc tcatgacgct
     2221 gaggctgtgg tccagtggct ccggagccac gaacttctct ctgttaaagc aagcaggaga
     2281 cgtggaagaa aaccccggtc ccatgaacaa gtgggttttc tgctgggtaa ccctttgtct
     2341 ccttactgta gagaccacac atggtgatgg tggcatcatt actcagacac ccaaattcct
     2401 gattggtcag gaagggcaaa aactgacctt gaaatgtcaa cagaatttca atcatgatac
     2461 aatgtactgg taccgacagg attcagggaa aggattgaga ctgatctact attcaataac
     2521 tgaaaacgat cttcaaaaag gcgatctatc tgaaggctat gatgcgtctc gcgagaagaa
     2581 gtcatctttt tctctcactg tgacatctgc ccagaagaac gagatggccg tttttctctg
     2641 tgccagcagt aaacaagaca cccagtactt tgggccaggc actcggctcc tcgtgttaga
     2701 agatctgaga aatgtgactc cacccaaggt ctccttgttt gagccatcaa aagcagagat
     2761 tgcaaacaaa caaaaggcta ccctcgtgtg cttggccagg ggcttcttcc ctgaccacgt
     2821 ggagctgagc tggtgggtga atggcaagga ggtccacagt ggggtcagca cggaccctca
     2881 ggcctacaag gagagcaatt atagctactg cctgagcagc cgcctgaggg tctctgctac
     2941 cttctggcac aatcctcgaa accacttccg ctgccaagtg cagttccatg ggctttcaga
     3001 ggaggacaag tggccagagg gctcacccaa acctgtcaca cagaacatca gtgcagaggc
     3061 ctggggccga gcagactgtg gaatcacttc agcatcctat catcaggggg ttctgtctgc
     3121 aaccatcctc tatgagatcc tactggggaa ggccacccta tatgctgtgc tggtcagtgg
     3181 cctggtgctg atggccatgg tcaagaaaaa aaattcctga gaattccgcc ccccccccct
     3241 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     3301 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     3361 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     3421 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     3481 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     3541 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     3601 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     3661 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     3721 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     3781 acacgatgat aatatggcca caaccatggg ccaccatgaa cccagccatc agcgtcgctc
     3841 tcctgctctc agtcttgcag gtgtcccgag ggcagaaggt gaccagcctg acagcctgcc
     3901 tggtgaacca aaaccttcgc ctggactgcc gccatgagaa taacaccaag gataactcca
     3961 tccagcatga gttcagcctg acccgagaga agaggaagca cgtgctctca ggcacccttg
     4021 ggatacccga gcacacgtac cgctcccgcg tcaccctctc caaccagccc tatatcaagg
     4081 tccttaccct agccaacttc accaccaagg atgagggcga ctacttttgt gagcttcgcg
     4141 tctcgggcgc gaatcccatg agctccaata aaagtatcag tgtgtataga gacaagctgg
     4201 tcaagtgtgg cggcataagc ctgctggttc agaacacatc ctggatgctg ctgctgctgc
     4261 tttccctctc cctcctccaa gccctggact tcatttctct gtgagtcgac ctgcagccaa
     4321 gcttatcgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     4381 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac
     4441 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg
     4501 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     4561 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     4621 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     4681 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     4741 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt
     4801 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     4861 ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc
     4921 tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac
     4981 atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt
     5041 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5101 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5161 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5221 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5281 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5341 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5401 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5461 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5521 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5581 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5641 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5701 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5761 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5821 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5881 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5941 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6001 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6061 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6121 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6181 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6241 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6301 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6361 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6421 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6481 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6541 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6601 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6661 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6721 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6781 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6841 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6901 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6961 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7021 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7081 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7141 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     7201 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     7261 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     7321 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     7381 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     7441 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     7501 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     7561 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     7621 ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     7681 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     7741 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     7801 ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac
     7861 aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata
     7921 gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa gggggg
//
