LOCUS       sequence-223050-        8052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of truncated Rag1 including the RING, E3 ubiquitin ligase
            (265-1008) and C-terminal region (384-1040).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8052)
  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 8052)
  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: UR1Ct (265-1040)
FEATURES             Location/Qualifiers
     source          1..8052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          88..1104
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1028..1050
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1112..1131
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1180..1189
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1192..1215
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             1222..2319
                     /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      2176..2185
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2293..2316
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2368..2391
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(4831..4850)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4896..4951
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4950..4969)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5312..5328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5312..5328)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5325..5347)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5336..5352
                     /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(5360..5366,5367..5384,5385..5390))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5405..5426
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5555..5572)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5726..6314)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5806..5825)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6485..7276,7277..7345))
                     /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     7108..7127
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7346..7450)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7481..7860
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7862..8052,1..87)
                     /label=chicken beta-actin promoter
ORIGIN
        1 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
       61 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
      121 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
      181 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
      241 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
      301 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
      361 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
      421 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
      481 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
      541 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
      601 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
      661 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
      721 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
      781 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
      841 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
      901 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
      961 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
     1021 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt
     1081 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
     1141 cattttggca aagaattgcg gccgtctcag gccgaattcg ccaccatggt gcatcaccat
     1201 caccatcacc atcatcgatc catgatcgaa gaaggtaaac tggtaatctg gattaacggc
     1261 gataaaggct ataacggtct cgctgaagtc ggtaagaaat tcgagaaaga taccggaatt
     1321 aaagtcaccg ttgagcatcc ggataaactg gaagagaaat tcccacaggt tgcggcaact
     1381 ggcgatggcc ctgacattat cttctgggca cacgaccgct ttggtggcta cgctcaatct
     1441 ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg acaagctgta tccgtttacc
     1501 tgggatgccg tacgttacaa cggcaagctg attgcttacc cgatcgctgt tgaagcgtta
     1561 tcgctgattt ataacaaaga tctgctgccg aacccgccaa aaacctggga agagatcccg
     1621 gcgctggata aagaactgaa agcgaaaggt aagagcgcgc tgatgttcaa cctgcaagaa
     1681 ccgtacttca cctggccgct gattgctgct gacgggggtt atgcgttcaa gtatgaaaac
     1741 ggcaagtacg acattaaaga cgtgggcgtg gataacgctg gcgcgaaagc gggtctgacc
     1801 ttcctggttg acctgattaa aaacaaacac atgaatgcag acaccgatta ctccatcgca
     1861 gaagctgcct ttaataaagg cgaaacagcg atgaccatca acggcccgtg ggcatggtcc
     1921 aacatcgaca ccagcaaagt gaattatggt gtaacggtac tgccgacctt caagggtcaa
     1981 ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta acgccgccag tccgaacaaa
     2041 gagctggcaa aagagttcct cgaaaactat ctgctgactg atgaaggtct ggaagcggtt
     2101 aataaagaca aaccgctggg tgccgtagcg ctgaagtctt acgaggaaga gttggcgaaa
     2161 gatccacgta ttgccgccac catggaaaac gcccagaaag gtgaaatcat gccgaacatc
     2221 ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg tgatcaacgc cgccagcggt
     2281 cgtcagactg tcgatgaagc cctgaaagac gcgcagacta attcgagctc gaacaacaac
     2341 aacaataaca ataacaacaa cctcgggctc gaagtcctat tccagggacc ggtgaactgc
     2401 agtaagattc atctcagtac caagcttctt gccgtggact tcccagcaca ctttgtgaaa
     2461 tccatctcct gccagatatg cgaacacatt ctggctgatc ccgtggagac cagctgcaag
     2521 catctattct gtaggatctg cattctcaga tgtctcaaag tcatgggcag ctattgtccc
     2581 tcttgccgat atccgtgctt ccctactgac ctggagagcc cagtgaagtc ctttctgaac
     2641 atcttgaatt ctctcatggt caagtgtccc gcgcaagatt gcaatgagga agtgagtctg
     2701 gaaaaatata accaccatgt gtcaagccac aaagaatcta aagagacttt ggtgcatatc
     2761 aataaagggg gacggcctcg ccagcatctc ctgtcactga cgagaagggc gcagaaacat
     2821 cggctgaggg agctcaagat tcaagtcaaa gaatttgctg acaaagaaga aggtggagat
     2881 gtgaaggctg tctgcttgac attgtttctc ctggcactga gggcgaggaa tgagcacagg
     2941 caagctgatg aattagaggc catcatgcaa ggcaggggct ccgggcttca accagctgtt
     3001 tgcttggcca tccgtgtcaa taccttcctc agctgtagcc aataccataa gatgtacagg
     3061 actgtgaaag ctatcactgg gaggcagatt tttcaacctt tgcatgctct tcggaatgcc
     3121 gagaaagtcc ttctgccagg ctaccatccc tttgagtggc agcccccact gaagaatgtg
     3181 tcctccagaa ctgatgttgg aattattgat gggctgtctg gacttgcctc ctctgtggat
     3241 gagtacccag tagataccat tgcgaagagg ttccgctacg actctgcttt ggtgtctgct
     3301 ttgatggaca tggaagaaga catcttggaa ggcatgagat cccaagatct tgatgactac
     3361 ctgaatggtc ccttcacagt ggtggtaaag gagtcttgcg atggaatggg ggatgtgagt
     3421 gagaagcacg ggagtgggcc cgcagttcca gaaaaggccg ttcgtttctc tttcacagtc
     3481 atgagaatta cgatagagca tggttcacag aacgtgaagg tgtttgagga acccaagccc
     3541 aattctgaac tgtgttgcaa gccgttgtgt cttatgctgg cagatgagtc tgaccatgag
     3601 acccttactg ctattctaag ccccctcatt gccgagaggg aggccatgaa gagcagtgaa
     3661 ttaacgctgg agatgggagg catccccagg acttttaaat tcatcttcag gggcactgga
     3721 tacgatgaaa aacttgtccg ggaagtagaa ggcttggaag cttctggctc agtctacatc
     3781 tgtacactct gtgacaccac ccgtttggaa gcctctcaga atcttgtctt ccactccata
     3841 accagaagcc acgccgagaa cctgcagcgc tatgaggtct ggcggtccaa tccgtatcat
     3901 gagtccgtgg aagagctccg ggaccgggtg aaaggggtct ctgccaaacc tttcatcgag
     3961 acagtccctt ccatagatgc gcttcactgt gacattggca atgcagctga attctataag
     4021 attttccagc tggagatagg ggaagtgtat aaacatccca atgcctctaa agaggaaagg
     4081 aagagatggc aggccacgct ggacaaacat ctccggaaaa ggatgaactt aaaaccaatc
     4141 atgaggatga atggcaactt tgcccggaag cttatgaccc aagagactgt agacgcagtt
     4201 tgtgagttaa ttccttctga ggagaggcat gaagctctca gggagctcat ggacctttac
     4261 ctgaagatga aacccgtgtg gcgctcttca tgtcccgcta aagagtgtcc agagtccctc
     4321 tgtcagtaca gtttcaactc acagcgtttc gcggaactcc tctccaccaa gttcaaatat
     4381 agatacgagg gcaaaatcac caattacttt cacaaaacct tggcacatgt ccctgaaatt
     4441 attgaaaggg atggctctat cggggcctgg gcaagtgagg gaaatgaatc gggtaacaag
     4501 ctgtttagac ggtttcggaa aatgaatgcc aggcagtcca agtgctatga gatggaagat
     4561 gtcctgaaac atcactggct gtatacttca aaatacctcc agaagtttat gaatgctcat
     4621 aacgcgttaa aaagctctgg gtttaccatg aactcaaagg agaccttagg ggaccctttg
     4681 ggcattgagg actctctgga aagccaagat tcaatggagt tttaataact cgagactagt
     4741 atcgcgataa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     4801 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
     4861 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     4921 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     4981 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     5041 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     5101 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     5161 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     5221 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     5281 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5341 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5401 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5461 gaaacctgtc gtgccagcgg atcgatccgc tgcattaatg aatcggccaa cgcgcgggga
     5521 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5581 tctttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5641 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5701 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5761 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5821 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5881 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5941 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6001 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6061 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6121 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6181 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6241 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6301 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6361 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6421 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6481 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6541 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6601 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6661 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6721 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6781 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6841 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6901 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6961 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7021 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7081 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7141 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7201 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7261 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7321 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7381 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7441 gggttccgcg cacatttccc cgaaaagtgc cacctgggtc gacattgatt attgactagt
     7501 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     7561 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     7621 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     7681 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     7741 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     7801 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     7861 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
     7921 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
     7981 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt
     8041 gcggcggcag cc
//
