LOCUS       sequence-382630-        8270 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 8270)
  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 8270)
  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-B
FEATURES             Location/Qualifiers
     source          1..8270
                     /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             2602..2658
                     /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    3605..4154
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3749..3766)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3976..3995
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4157..4159,4160..4162,4163..4876)
                     /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(4157..4159,4160..4162,4163..4873)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4202..4223)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4463..4482)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4810..4831
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5038..5552
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5714..5730
                     /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(5738..5744,5745..5762,5763..5768))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5783..5804
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5921..5938)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6092..6680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6172..6191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6851..7642,7643..7711))
                     /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     7474..7493
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7712..7816)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7884..7902
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7940..7962)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8062..8081
                     /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 gagaggagat
     1861 caggtggagc agagtccttc agccctgagc ctccacgagg gaaccggttc tgctctgaga
     1921 tgcaatttta ctaccaccat gagggctgtg cagtggttcc gaaagaattc caggggcagc
     1981 ctcatcaatc tgttctactt ggcttcagga acaaaggaga atgggaggct aaagtcagca
     2041 tttgattcta aggagcgcta cagcaccctg cacatcaggg atgcccagct ggaggactca
     2101 ggcacttact tctgtgctgc tggggcatct tctggcagct ggcaactcat ctttggatct
     2161 ggaacccaac tgacagttat gcctgacatc cagaacccag aacctgctgt gtaccagtta
     2221 aaagatcctc ggtctcagga cagcaccctc tgcctgttca ccgactttga ctcccaaatc
     2281 aatgtgccga aaaccatgga atctggaacg ttcatcactg acaaaactgt gctggacatg
     2341 aaagctatgg attccaagag caatggggcc attgcctgga gcaaccagac aagcttcacc
     2401 tgccaagata tcttcaaaga gaccaacgcc acctacccca gttcagacgt tccctgtgat
     2461 gccacgttga ctgagaaaag ctttgaaaca gatatgaacc taaactttca aaacctgtca
     2521 gttatgggac tccgaatcct cctgctgaaa gtagccggat ttaacctgct catgacgctg
     2581 aggctgtggt ccggctccgg agccacgaac ttctctctgt taaagcaagc aggagacgtg
     2641 gaagaaaacc ccggtcccat gggctccagg ctctttctgg tcttgagcct cctgtgtaca
     2701 aaacacatgg aggctgcagt cacccaaagc cctagaaaca aggtgacagt aacaggagga
     2761 aacgtgacat tgagctgtcg ccagactaat agccacaact acatgtactg gtatcggcag
     2821 gacactgggc atgggctgag gctgatccat tactcatatg gtgctggcaa ccttcaaata
     2881 ggagatgtcc ctgatgggta caaggccacc agaacaacgc aagaagactt cttcctcctg
     2941 ctggaattgg cttctccctc tcagacatct ttgtacttct gtgccagcag cgcgggactg
     3001 gggagctcct atgaacagta cttcggtccc ggcaccaggc tcacggtttt agaggatctg
     3061 agaaatgtga ctccacccaa ggtctccttg tttgagccat caaaagcaga gattgcaaac
     3121 aaacaaaagg ctaccctcgt gtgcttggcc aggggcttct tccctgacca cgtggagctg
     3181 agctggtggg tgaatggcaa ggaggtccac agtggggtca gcacggaccc tcaggcctac
     3241 aaggagagca attatagcta ctgcctgagc agccgcctga gggtctctgc taccttctgg
     3301 cacaatcctc gaaaccactt ccgctgccaa gtgcagttcc atgggctttc ggaggaggac
     3361 aagtggccag agggctcacc caaacctgtc acacagaaca tcagtgcaga ggcctggggc
     3421 cgagcagact gtggaatcac ttcagcatcc tatcatcagg gggttctgtc tgcaaccatc
     3481 ctctatgaga tcctactggg gaaggccacc ctatatgctg tgctggtcag tggcctggtg
     3541 ctgatggcca tggtcaagag aaaaaattcc tgactcgagc gggatcaatt ccgccccccc
     3601 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta
     3661 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
     3721 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
     3781 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc
     3841 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
     3901 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
     3961 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
     4021 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt
     4081 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     4141 aaaacacgat aataccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     4201 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     4261 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     4321 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     4381 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     4441 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     4501 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     4561 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     4621 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     4681 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     4741 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     4801 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     4861 cgagctgtac aagtaaagcg gccgcgactc tagagtcgac ctgcaggcat gcaagcttca
     4921 ggtagccggc taacgttaac aaccggtacc tctagaacta tagctagcat gcgcaaattt
     4981 aaagcgctga tatcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
     5041 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
     5101 aaaatacata actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga
     5161 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     5221 agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc
     5281 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     5341 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     5401 cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt atccaataaa
     5461 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     5521 tgattgacta cccgtcagcg ggggtctttc atgggtaaca gtttcttgaa gttggagaac
     5581 aacattctga gggtaggagt cgaatattaa gtaatcctga ctcaattagc cactgttttg
     5641 aatccacata ctccaatact cctgaaatag ttcattatgg acagcgcaga aagagctggg
     5701 gagaattgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5761 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5821 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5881 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5941 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6001 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6061 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6121 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6181 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6241 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6301 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6361 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6421 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6481 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6541 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6601 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6661 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6721 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6781 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6841 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6901 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6961 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7021 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7081 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7141 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7201 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7261 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7321 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7381 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7441 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7501 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7561 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7621 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7681 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7741 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7801 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     7861 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     7921 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     7981 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     8041 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     8101 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     8161 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     8221 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
//
