LOCUS       sequence-223073-        7027 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expression of full length Rag2 with T490A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7027)
  AUTHORS   Grundy GJ, Yang W, Gellert M
  TITLE     Autoinhibition of DNA cleavage mediated by RAG1 and RAG2 is overcome
            by an epigenetic signal in V(D)J recombination.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Dec 28;107(52):22487-92. doi: 
            10.1073/pnas.1014958107. Epub 2010 Dec 13.
  PUBMED    21149691
REFERENCE   2  (bases 1 to 7027)
  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: 
            "10.1073/pnas.1014958107"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2010-12-28- 28"; volume: "107"; issue: "52"; pages: 
            "22487-92"
COMMENT     Sequence Label: Rag2 T490A
FEATURES             Location/Qualifiers
     source          1..7027
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     940..962
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1024..1043
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1092..1101
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1104..1127
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             1134..2231
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      2088..2097
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2205..2228
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2280..2303
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3996..4015)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4061..4116
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4115..4134)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4477..4493)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4477..4493)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4490..4512)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4501..4517
                     /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(4525..4531,4532..4549,4550..4555))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4570..4591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4720..4737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4891..5479)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4971..4990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5650..6441,6442..6510))
                     /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     6273..6292
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6511..6615)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6646..7025
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
       61 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      121 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      181 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      241 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      301 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
      361 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
      421 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
      481 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
      541 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
      601 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
      661 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
      721 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
      781 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
      841 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
      901 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
      961 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1021 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg cggccgtctc
     1081 aggccgaatt cgccaccatg gtgcatcacc atcaccatca ccatcatcga tccatgatcg
     1141 aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt ctcgctgaag
     1201 tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat ccggataaac
     1261 tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt atcttctggg
     1321 cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc accccggaca
     1381 aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac aacggcaagc
     1441 tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa gatctgctgc
     1501 cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg aaagcgaaag
     1561 gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg ctgattgctg
     1621 ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa gacgtgggcg
     1681 tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt aaaaacaaac
     1741 acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa ggcgaaacag
     1801 cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa gtgaattatg
     1861 gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga
     1921 gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc ctcgaaaact
     1981 atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag
     2041 cgctgaagtc ttacgaggaa gagttggcga aagatccacg tattgccgcc accatggaaa
     2101 acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc tggtatgccg
     2161 tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag
     2221 acgcgcagac taattcgagc tcgaacaaca acaacaataa caataacaac aacctcgggc
     2281 tcgaagtcct attccaggga ccggtgtccc tgcagatggt aacagtgggt cataacatag
     2341 ccttaattca accaggcttc tcacttatga attttgatgg ccaagttttc ttctttggcc
     2401 agaaaggctg gcctaagaga tcctgtccta ctggagtctt tcattttgat ataaaacaaa
     2461 atcatctcaa actgaagcct gcaatcttct ctaaagattc ctgctacctc ccacctcttc
     2521 gttatccagc tacttgctca tacaaaggca gcatagactc tgacaagcat caatatatca
     2581 ttcacggagg gaaaacacca aacaatgagc tttccgataa gatttatatc atgtctgtcg
     2641 cttgcaagaa taacaaaaaa gttactttcc gttgcacaga gaaagactta gtaggagatg
     2701 tccctgaacc cagatacggc cattccattg acgtggtgta tagtcgaggg aaaagcatgg
     2761 gtgttctctt tggaggacgt tcatacatgc cttctaccca gagaaccaca gaaaaatgga
     2821 atagtgtagc tgactgccta ccccatgttt tcttgataga ttttgaattt gggtgtgcta
     2881 catcatatat tctcccagaa cttcaggatg ggctgtcttt tcatgtttct attgccagaa
     2941 acgataccgt ttatattttg ggaggacact cacttgccag taatatacgc cctgctaact
     3001 tgtatagaat aagagtggac cttcccctgg gtaccccagc agtgaattgc acagtcttgc
     3061 caggaggaat ctctgtctcc agtgcaatcc tcactcaaac aaacaatgat gaatttgtta
     3121 ttgtgggtgg ttatcagctg gaaaatcaga aaaggatggt ctgcagcctt gtctctctag
     3181 gggacaacac gattgaaatc agtgagatgg agactcctga ctggacctca gatattaagc
     3241 atagcaaaat atggtttgga agcaacatgg gaaacgggac tattttcctt ggcataccag
     3301 gagacaataa gcaggctatg tcagaagcat tctatttcta tactttgaga tgctctgaag
     3361 aggatttgag tgaagatcag aaaattgtct ccaacagtca gacatcaaca gaagatcctg
     3421 gggactccac tccctttgaa gactcagagg aattttgttt cagtgctgaa gcaaccagtt
     3481 ttgatggtga cgatgaattt gacacctaca atgaagatga tgaagatgac gagtctgtaa
     3541 ccggctactg gataacatgt tgccctactt gtgatgttga catcaatacc tgggttccgt
     3601 tctattcaac ggagctcaat aaacccgcca tgatctattg ttctcatggg gatgggcact
     3661 gggtacatgc ccagtgcatg gatttggaag aacgcacact catccacttg tcagaaggaa
     3721 gcaacaagta ttattgcaat gaacatgtac agatagcaag agcattgcaa gctcccaaaa
     3781 gaaacccccc cttacaaaaa cctccaatga aatccctcca caaaaaaggc tctgggaaag
     3841 tcttgactcc tgccaagaaa tccttcctta gaagactgtt tgattaataa taactcgaga
     3901 ctagtatcgc gataattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     3961 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     4021 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     4081 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     4141 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     4201 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     4261 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     4321 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     4381 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     4441 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4501 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4561 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4621 gtcgggaaac ctgtcgtgcc agcggatcga tccgctgcat taatgaatcg gccaacgcgc
     4681 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4741 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4801 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4861 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4921 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4981 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5041 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5101 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5161 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5221 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5281 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5341 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5401 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5461 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5521 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5581 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5641 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5701 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5761 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5821 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5881 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5941 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6001 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6061 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6121 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6181 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6241 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6301 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6361 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6421 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6481 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6541 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6601 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgacat tgattattga
     6661 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6721 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     6781 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     6841 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6901 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     6961 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     7021 ccatggt
//
