LOCUS       sequence-246262-        7964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the Helicobacter hepaticus specific TCRa/b HH5-1 in T cell 
            hybridoma cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7964)
  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 7964)
  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-HH5-1
FEATURES             Location/Qualifiers
     source          1..7964
                     /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             2234..2290
                     /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    3213..3780
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3230..3782
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3374..3391)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3601..3620
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3804..4292
                     /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             4359..4873
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5035..5051
                     /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(5059..5065,5066..5083,5084..5089))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5104..5125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5242..5259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5413..6001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5493..5512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6172..6963,6964..7032))
                     /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     6795..6814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7033..7137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7205..7223
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7261..7283)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7383..7402
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7596..7618
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7746..7765
                     /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 tgcacagcct
     1441 cctggggttg ttgttgtggc tgcaactgac aagggtgaat agtcaactag cagaagaaaa
     1501 ttcgtgggcc ctgagcgtcc acgagggtga aagtgtcacg gtgaattgta gttacaagac
     1561 atccataact gccctacagt ggtacagaca gaagtcaggc aaaggccctg cccagctaat
     1621 cttaatacgt tcaaatgaga gagagaagcg caatggaaga ctcagagcca cccttgacac
     1681 ctccagccag agcagctcct tgtccatcac tgctactcgg tgtgaagaca ccgctgtgta
     1741 cttctgtgct actgatgata ccaatacagg caaattaacc tttggggatg ggaccgtgct
     1801 cacagtgaag ccaaacatcc agaacccaga acctgctgtg taccagttaa aagatcctcg
     1861 gtctcaggac agcaccctct gcctgttcac cgactttgac tcccaaatca atgtgccgaa
     1921 aaccatggaa tctggaacgt tcatcactga caaaactgtg ctggacatga aagctatgga
     1981 ttccaagagc aatggggcca ttgcctggag caaccagaca agcttcacct gccaagatat
     2041 cttcaaagag accaacgcca cctaccccag ttcagacgtt ccctgtgatg ccacgttgac
     2101 tgagaaaagc tttgaaacag atatgaacct aaactttcaa aacctgtcag ttatgggact
     2161 ccgaatcctc ctgctgaaag tagccggatt taacctgctc atgacgctga ggctgtggtc
     2221 cagtggctcc ggagccacga acttctctct gttaaagcaa gcaggagacg tggaagaaaa
     2281 ccccggtccc atgggctcca ggctcttctt cgtgctctcc agtctcctgt gttcaaaaca
     2341 catggaggct gcagtcaccc aaagcccaag aaacaaggtg gcagtaacag gaggaaaggt
     2401 gacattgagc tgtaatcaga ctaataacca caacaacatg tactggtatc ggcaggacac
     2461 ggggcatggg ctgaggctga tccattattc atatggtgct ggcagcactg agaaaggaga
     2521 tatccctgat ggatacaagg cctccagacc aagccaagag aacttctccc tcattctgga
     2581 gttggctacc ccctctcaga catcagtgta cttctgtgcc agcggtggga cagggggcaa
     2641 ccaagacacc cagtactttg ggccaggcac tcggctcctc gtgttagaag atctgagaaa
     2701 tgtgactcca cccaaggtct ccttgtttga gccatcaaaa gcagagattg caaacaaaca
     2761 aaaggctacc ctcgtgtgct tggccagggg cttcttccct gaccacgtgg agctgagctg
     2821 gtgggtgaat ggcaaggagg tccacagtgg ggtcagcacg gaccctcagg cctacaagga
     2881 gagcaattat agctactgcc tgagcagccg cctgagggtc tctgctacct tctggcacaa
     2941 tcctcgaaac cacttccgct gccaagtgca gttccatggg ctttcagagg aggacaagtg
     3001 gccagagggc tcacccaaac ctgtcacaca gaacatcagt gcagaggcct ggggccgagc
     3061 agactgtgga atcacttcag catcctatca tcagggggtt ctgtctgcaa ccatcctcta
     3121 tgagatccta ctggggaagg ccaccctata tgctgtgctg gtcagtggcc tggtgctgat
     3181 ggccatggtc aagaaaaaaa attcctgaga attccgcccc ccccccctaa cgttactggc
     3241 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     3301 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     3361 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     3421 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     3481 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     3541 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     3601 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     3661 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     3721 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     3781 tatggccaca accatgggcc accatgaacc cagccatcag cgtcgctctc ctgctctcag
     3841 tcttgcaggt gtcccgaggg cagaaggtga ccagcctgac agcctgcctg gtgaaccaaa
     3901 accttcgcct ggactgccgc catgagaata acaccaagga taactccatc cagcatgagt
     3961 tcagcctgac ccgagagaag aggaagcacg tgctctcagg cacccttggg atacccgagc
     4021 acacgtaccg ctcccgcgtc accctctcca accagcccta tatcaaggtc cttaccctag
     4081 ccaacttcac caccaaggat gagggcgact acttttgtga gcttcgcgtc tcgggcgcga
     4141 atcccatgag ctccaataaa agtatcagtg tgtatagaga caagctggtc aagtgtggcg
     4201 gcataagcct gctggttcag aacacatcct ggatgctgct gctgctgctt tccctctccc
     4261 tcctccaagc cctggacttc atttctctgt gagtcgacct gcagccaagc ttatcgataa
     4321 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     4381 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat
     4441 agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata
     4501 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     4561 tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     4621 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     4681 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
     4741 ccgatagact gcgtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
     4801 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
     4861 gggggtcttt catgggtaac agtttcttga agttggagaa caacattctg agggtaggag
     4921 tcgaatatta agtaatcctg actcaattag ccactgtttt gaatccacat actccaatac
     4981 tcctgaaata gttcattatg gacagcgcag aaagagctgg ggagaattgt gaaattgtta
     5041 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5101 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5161 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5221 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5281 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5341 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5401 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5461 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5521 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5581 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5641 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5701 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5761 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5821 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5881 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5941 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6001 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6061 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6241 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6301 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6361 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6421 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6481 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6541 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6601 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6661 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6721 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6781 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6841 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6901 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6961 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7021 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7081 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7141 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     7201 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     7261 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     7321 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     7381 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     7441 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     7501 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     7561 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     7621 ggcgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     7681 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     7741 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     7801 gccacgatgc gtccggcgta gaggcgatta gtccaatttg ttaaagacag gatatcagtg
     7861 gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca
     7921 tagataaaat aaaagatttt atttagtctc cagaaaaagg gggg
//
