LOCUS       sequence-246716-        7961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the Helicobacter hepaticus specific TCRa/b HH7-8 in T cell 
            hybridoma cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7961)
  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 7961)
  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-HH7-8
FEATURES             Location/Qualifiers
     source          1..7961
                     /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    3210..3777
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3227..3779
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3371..3388)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3598..3617
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3801..4289
                     /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             4356..4870
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5032..5048
                     /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(5056..5062,5063..5080,5081..5086))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5101..5122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5239..5256)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5410..5998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5490..5509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6169..6960,6961..7029))
                     /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     6792..6811
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7030..7134)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7202..7220
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7258..7280)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7380..7399
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7593..7615
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7743..7762
                     /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 tgctcctggc
     1441 gctcctccca gtgctgggga tacactttgt cctgagagat gcccaagctc agtcagtgac
     1501 gcagcccgat gctcgcgtca ctgtctctga aggagcctct ctgcagctga gatgcaagta
     1561 ttcctcctct gggacacctt atctgttctg gtatgtccag tacccgcggc aggggctgca
     1621 gctgctcctc aagtactatt ccggagaccc agtggttcaa ggagtgaatg gctttgaggc
     1681 tgagttcagc aagagtaact cttccttcca cctgcggaaa gcctctgtgc actggagcga
     1741 ctcggctgtg tacttctgtg ttctgagcgc aggaggaaat gagaaaataa cttttggggc
     1801 tggaaccaaa ctcaccatta aacccaacat 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 ccatgctgta ctctctcctt gcctttctcc tgggcatgtt
     2341 cttgggtgtt agtgctcaga ctatccatca atggccagtt gccgagatca aggctgtggg
     2401 cagcccactg tctctggggt gtaccataaa ggggaaatca agccctaacc tctactggta
     2461 ctggcaggcc acaggaggca ccctccagca actcttctac tctattactg ttggccaggt
     2521 agagtcggtg gtgcaactga acctctcagc ttccaggccg aaggacgacc aattcatcct
     2581 aagcacggag aagctgcttc tcagccactc tggcttctac ctctgtgcct ggagggacag
     2641 aacagaagtc ttctttggta aaggaaccag actcacagtt gtagaagatc tgagaaatgt
     2701 gactccaccc aaggtctcct tgtttgagcc atcaaaagca gagattgcaa acaaacaaaa
     2761 ggctaccctc gtgtgcttgg ccaggggctt cttccctgac cacgtggagc tgagctggtg
     2821 ggtgaatggc aaggaggtcc acagtggggt cagcacggac cctcaggcct acaaggagag
     2881 caattatagc tactgcctga gcagccgcct gagggtctct gctaccttct ggcacaatcc
     2941 tcgaaaccac ttccgctgcc aagtgcagtt ccatgggctt tcagaggagg acaagtggcc
     3001 agagggctca cccaaacctg tcacacagaa catcagtgca gaggcctggg gccgagcaga
     3061 ctgtggaatc acttcagcat cctatcatca gggggttctg tctgcaacca tcctctatga
     3121 gatcctactg gggaaggcca ccctatatgc tgtgctggtc agtggcctgg tgctgatggc
     3181 catggtcaag aaaaaaaatt cctgagaatt ccgccccccc cccctaacgt tactggccga
     3241 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     3301 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     3361 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     3421 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     3481 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     3541 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     3601 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     3661 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     3721 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     3781 ggccacaacc atgggccacc atgaacccag ccatcagcgt cgctctcctg ctctcagtct
     3841 tgcaggtgtc ccgagggcag aaggtgacca gcctgacagc ctgcctggtg aaccaaaacc
     3901 ttcgcctgga ctgccgccat gagaataaca ccaaggataa ctccatccag catgagttca
     3961 gcctgacccg agagaagagg aagcacgtgc tctcaggcac ccttgggata cccgagcaca
     4021 cgtaccgctc ccgcgtcacc ctctccaacc agccctatat caaggtcctt accctagcca
     4081 acttcaccac caaggatgag ggcgactact tttgtgagct tcgcgtctcg ggcgcgaatc
     4141 ccatgagctc caataaaagt atcagtgtgt atagagacaa gctggtcaag tgtggcggca
     4201 taagcctgct ggttcagaac acatcctgga tgctgctgct gctgctttcc ctctccctcc
     4261 tccaagccct ggacttcatt tctctgtgag tcgacctgca gccaagctta tcgataaaat
     4321 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
     4381 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
     4441 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
     4501 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     4561 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     4621 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     4681 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     4741 atagactgcg tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     4801 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     4861 ggtctttcat gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg
     4921 aatattaagt aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc
     4981 tgaaatagtt cattatggac agcgcagaaa gagctgggga gaattgtgaa attgttatcc
     5041 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5101 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5161 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5221 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5281 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5341 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5401 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5461 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5521 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5581 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5641 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5701 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5761 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5821 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5881 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5941 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6001 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6061 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6121 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6181 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6241 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6301 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6361 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6421 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6481 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6541 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6601 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6661 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6721 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6781 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6841 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6901 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6961 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7021 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7081 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7141 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     7201 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     7261 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     7321 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     7381 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     7441 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     7501 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     7561 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     7621 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     7681 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     7741 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     7801 acgatgcgtc cggcgtagag gcgattagtc caatttgtta aagacaggat atcagtggtc
     7861 caggctctag ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag
     7921 ataaaataaa agattttatt tagtctccag aaaaaggggg g
//
