LOCUS       sequence-382627-        8288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of candidate orphan TCR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8288)
  AUTHORS   Akama-Garren EH, Yin X, Prestwood TR, Ma M, Utz PJ, Carroll MC
  TITLE     T cell help shapes B cell tolerance.
  JOURNAL   Sci Immunol. 2024 Feb 16;9(92):eadj7029. doi: 
            10.1126/sciimmunol.adj7029. Epub 2024 Feb 16.
  PUBMED    38363829
REFERENCE   2  (bases 1 to 8288)
  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"; journalName: "Sci 
            Immunol."; date: "2024-02-16"; volume: "9(92):eadj7029. doi"; pages:
            " 10.1126/sciimmunol.adj7029. Epub 2024 Feb 16"
COMMENT     Sequence Label: pMIGII-TCR-G
FEATURES             Location/Qualifiers
     source          1..8288
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..27
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     155..174
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             374..890
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    954..1295
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1362..1778
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1708..1730
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1746..1762
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             2617..2673
                     /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    3623..4172
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3767..3784)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3994..4013
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4175..4177,4178..4180,4181..4894)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4175..4177,4178..4180,4181..4891)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4220..4241)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4481..4500)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4828..4849
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5056..5570
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5732..5748
                     /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(5756..5762,5763..5780,5781..5786))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5801..5822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5939..5956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6110..6698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6190..6209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6869..7660,7661..7729))
                     /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     7492..7511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7730..7834)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7902..7920
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7958..7980)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8080..8099
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg
       61 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
      121 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
      181 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggcgattag
      241 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc
      301 accagctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc
      361 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
      421 cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg
      481 aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
      541 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag
      601 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
      661 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
      721 aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta
      781 cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggactc gctgatcctt
      841 gggagggtct cctcagattg attgactgcc cacctcgggg gtctttcatt tggaggttcc
      901 accgagattt ggagacccct gcccagggac caccgacccc cccgccggga ggtaagctgg
      961 ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt
     1021 tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
     1081 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
     1141 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
     1201 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
     1261 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
     1321 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
     1381 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1441 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1501 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1561 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1621 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1681 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1741 cctcccttta tccagccctc actccttctc taggcgccgg aattcagatc taccatgcgt
     1801 cctgtcacct gctcagttct tgtgctcctc ctaatgctca ggaggagcaa tggcgatgga
     1861 gactccgtga cccagacaga aggcctggtc actctcacag aagggttgcc tgtgatgctg
     1921 aactgcacct atcagactat ttactcaaat cctttccttt tctggtatgt gcaacatctc
     1981 aatgaatccc ctcggctact cctgaagagc ttcacagaca acaagaggac cgagcaccaa
     2041 gggttccacg ccactctcca taagagcagc agctccttcc atctgcagaa gtcctcagcg
     2101 cagctgtcag actctgccct gtactactgt gctctgagtg acgaggacta tgcaaacaag
     2161 atgatctttg gcttgggaac cattttgaga gtcagacctc acatccagaa cccagaacct
     2221 gctgtgtacc agttaaaaga tcctcggtct caggacagca ccctctgcct gttcaccgac
     2281 tttgactccc aaatcaatgt gccgaaaacc atggaatctg gaacgttcat cactgacaaa
     2341 actgtgctgg acatgaaagc tatggattcc aagagcaatg gggccattgc ctggagcaac
     2401 cagacaagct tcacctgcca agatatcttc aaagagacca acgccaccta ccccagttca
     2461 gacgttccct gtgatgccac gttgactgag aaaagctttg aaacagatat gaacctaaac
     2521 tttcaaaacc tgtcagttat gggactccga atcctcctgc tgaaagtagc cggatttaac
     2581 ctgctcatga cgctgaggct gtggtccggc tccggagcca cgaacttctc tctgttaaag
     2641 caagcaggag acgtggaaga aaaccccggt cccatgagct gcaggcttct cctctatgtt
     2701 tccctatgtc ttgtggaaac agcactcatg aacactaaaa ttactcagtc accaagatat
     2761 ctaatcctgg gaagagcaaa taagtctttg gaatgtgagc aacatctggg acataatgct
     2821 atgtactggt ataaacagag cgctgagaag ccgccagagc tcatgtttct ctacaatctt
     2881 aaacagttga ttcgaaatga gacggtgccc agtcgtttta tacctgaatg cccagacagc
     2941 tccaagctac ttttacatat atctgccgtg gatccagaag actcagctgt ctatttttgt
     3001 gccagcagcc aagggggggc tgatcaagac acccagtact ttgggccagg cactcggctc
     3061 ctcgtgttag aggatctgag aaatgtgact ccacccaagg tctccttgtt tgagccatca
     3121 aaagcagaga ttgcaaacaa acaaaaggct accctcgtgt gcttggccag gggcttcttc
     3181 cctgaccacg tggagctgag ctggtgggtg aatggcaagg aggtccacag tggggtcagc
     3241 acggaccctc aggcctacaa ggagagcaat tatagctact gcctgagcag ccgcctgagg
     3301 gtctctgcta ccttctggca caatcctcga aaccacttcc gctgccaagt gcagttccat
     3361 gggctttcgg aggaggacaa gtggccagag ggctcaccca aacctgtcac acagaacatc
     3421 agtgcagagg cctggggccg agcagactgt ggaatcactt cagcatccta tcatcagggg
     3481 gttctgtctg caaccatcct ctatgagatc ctactgggga aggccaccct atatgctgtg
     3541 ctggtcagtg gcctggtgct gatggccatg gtcaagagaa aaaattcctg actcgagcgg
     3601 gatcaattcc gccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     3661 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     3721 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     3781 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     3841 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     3901 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     3961 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     4021 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     4081 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     4141 gacgtggttt tcctttgaaa aacacgataa taccatggtg agcaagggcg aggagctgtt
     4201 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     4261 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     4321 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     4381 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     4441 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     4501 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     4561 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     4621 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     4681 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     4741 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     4801 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4861 gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gagtcgacct
     4921 gcaggcatgc aagcttcagg tagccggcta acgttaacaa ccggtacctc tagaactata
     4981 gctagcatgc gcaaatttaa agcgctgata tcgataaaat aaaagatttt atttagtctc
     5041 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     5101 ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag
     5161 gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     5221 cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc agtttctaga
     5281 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     5341 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     5401 aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt
     5461 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     5521 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt
     5581 ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact
     5641 caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatagtt cattatggac
     5701 agcgcagaaa gagctgggga gaattgtgaa attgttatcc gctcacaatt ccacacaaca
     5761 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5821 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5881 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5941 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6001 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6061 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6121 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6181 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6241 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6301 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6361 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6421 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6481 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6541 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6601 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6661 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6721 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6781 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6841 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6901 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6961 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7021 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7081 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7141 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7201 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7261 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7321 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7381 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7441 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7501 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7561 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7621 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7681 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7741 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7801 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7861 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     7921 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     7981 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     8041 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     8101 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     8161 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     8221 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     8281 cgccaggg
//
