LOCUS       sequence-382672-        8273 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 8273)
  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 8273)
  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-A
FEATURES             Location/Qualifiers
     source          1..8273
                     /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             2605..2661
                     /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    3608..4157
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3752..3769)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3979..3998
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4160..4162,4163..4165,4166..4879)
                     /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(4160..4162,4163..4165,4166..4876)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4205..4226)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4466..4485)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4813..4834
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5041..5555
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5717..5733
                     /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(5741..5747,5748..5765,5766..5771))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5786..5807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5924..5941)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6095..6683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6175..6194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6854..7645,7646..7714))
                     /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     7477..7496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7715..7819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7887..7905
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7943..7965)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8065..8084
                     /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 taccatgcag
     1801 aggaacctgg gagctgtgct ggggattctg tgggtgcaga tttgctgggt gagcggagat
     1861 aaggtgaaac aaagtccctc agcgctgagt ctccaagaag gaaccaattc tgctctgaga
     1921 tgcaattttt ctatcgccgc gacaactgtg cagtggttcc tacagaatcc caggggcagc
     1981 ctcatcaatc ttttttacct ggttccagga acaaaggaga atgggaggtt aaagtcagca
     2041 ttcgattcta aggagagcta cagcaccctg cacatcaggg atgcccagct ggaggactca
     2101 ggcacttact tctgtgctgc tgggtctccc aactacagca acaacagact tactttgggg
     2161 aagggaaccc aggtggtggt gttaccaaac atccagaacc cagaacctgc tgtgtaccag
     2221 ttaaaagatc ctcggtctca ggacagcacc ctctgcctgt tcaccgactt tgactcccaa
     2281 atcaatgtgc cgaaaaccat ggaatctgga acgttcatca ctgacaaaac tgtgctggac
     2341 atgaaagcta tggattccaa gagcaatggg gccattgcct ggagcaacca gacaagcttc
     2401 acctgccaag atatcttcaa agagaccaac gccacctacc ccagttcaga cgttccctgt
     2461 gatgccacgt tgactgagaa aagctttgaa acagatatga acctaaactt tcaaaacctg
     2521 tcagttatgg gactccgaat cctcctgctg aaagtagccg gatttaacct gctcatgacg
     2581 ctgaggctgt ggtccggctc cggagccacg aacttctctc tgttaaagca agcaggagac
     2641 gtggaagaaa accccggtcc catgggctgt aggctcctaa gctgtgtggc cttctgcctc
     2701 ttgggaatag gccctttgga gacggctgtt ttccagactc caaactatca tgtcacacag
     2761 gtgggaaatg aagtgtcttt caattgtaag caaactctgg gccacgatac tatgtattgg
     2821 tacaagcaag actctaagaa attgctgaag attatgttta gctacaataa taagcaactc
     2881 attgtaaacg aaacagttcc aaggcgcttc tcacctcagt cttcagataa agctcatttg
     2941 aatcttcgaa tcaagtctgt agagccggag gactctgctg tgtatctctg tgccagcagc
     3001 agggactggg gggggctaga gtacttcggt cccggcacca ggctcacggt tttagaggat
     3061 ctgagaaatg tgactccacc caaggtctcc ttgtttgagc catcaaaagc agagattgca
     3121 aacaaacaaa aggctaccct cgtgtgcttg gccaggggct tcttccctga ccacgtggag
     3181 ctgagctggt gggtgaatgg caaggaggtc cacagtgggg tcagcacgga ccctcaggcc
     3241 tacaaggaga gcaattatag ctactgcctg agcagccgcc tgagggtctc tgctaccttc
     3301 tggcacaatc ctcgaaacca cttccgctgc caagtgcagt tccatgggct ttcggaggag
     3361 gacaagtggc cagagggctc acccaaacct gtcacacaga acatcagtgc agaggcctgg
     3421 ggccgagcag actgtggaat cacttcagca tcctatcatc agggggttct gtctgcaacc
     3481 atcctctatg agatcctact ggggaaggcc accctatatg ctgtgctggt cagtggcctg
     3541 gtgctgatgg ccatggtcaa gagaaaaaat tcctgactcg agcgggatca attccgcccc
     3601 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     3661 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     3721 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     3781 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     3841 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     3901 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     3961 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     4021 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     4081 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     4141 tgaaaaacac gataatacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     4201 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     4261 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     4321 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     4381 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4441 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4501 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4561 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     4621 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     4681 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4741 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4801 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4861 ggacgagctg tacaagtaaa gcggccgcga ctctagagtc gacctgcagg catgcaagct
     4921 tcaggtagcc ggctaacgtt aacaaccggt acctctagaa ctatagctag catgcgcaaa
     4981 tttaaagcgc tgatatcgat aaaataaaag attttattta gtctccagaa aaagggggga
     5041 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     5101 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
     5161 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     5221 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
     5281 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     5341 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     5401 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat
     5461 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     5521 gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag
     5581 aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt
     5641 ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct
     5701 ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5761 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5821 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5881 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5941 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6001 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6061 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6121 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6181 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6241 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6301 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6361 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6421 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6481 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6541 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6601 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6661 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6721 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6781 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6841 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6901 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6961 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7021 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7081 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7141 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7201 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7261 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7321 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7381 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7441 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7501 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7561 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7621 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7681 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7741 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7801 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     7861 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     7921 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     7981 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     8041 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     8101 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     8161 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     8221 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca ggg
//
