LOCUS       Exported                7593 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  expression of truncated Rag1.
ACCESSION   .
VERSION     .
KEYWORDS    coreR1 (384-1008)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7593)
  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 7593)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7593
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     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(4372..4391)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4437..4492
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4491..4510)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4853..4869)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4853..4869)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4866..4888)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4877..4893
                     /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(4901..4931)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4946..4967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5096..5113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5267..5855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5347..5366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6026..6886)
                     /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     6649..6668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6887..6991)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7022..7401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7403..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 ggtgcatatc
     2401 aataaagggg gacggcctcg ccagcatctc ctgtcactga cgagaagggc gcagaaacat
     2461 cggctgaggg agctcaagat tcaagtcaaa gaatttgctg acaaagaaga aggtggagat
     2521 gtgaaggctg tctgcttgac attgtttctc ctggcactga gggcgaggaa tgagcacagg
     2581 caagctgatg aattagaggc catcatgcaa ggcaggggct ccgggcttca accagctgtt
     2641 tgcttggcca tccgtgtcaa taccttcctc agctgtagcc aataccataa gatgtacagg
     2701 actgtgaaag ctatcactgg gaggcagatt tttcaacctc tgcatgctct tcggaatgcc
     2761 gagaaagtcc ttctgccagg ctaccatccc tttgagtggc agcccccact gaagaatgtg
     2821 tcctccagaa ctgatgttgg aattattgat gggctgtctg gacttgcctc ctctgtggat
     2881 gagtacccag tagataccat tgcgaagagg ttccgctacg actctgcttt ggtgtctgct
     2941 ttgatggaca tggaagaaga catcttggaa ggcatgagat cccaagatct tgatgactac
     3001 ctgaatggtc ccttcacagt ggtggtaaag gagtcttgcg atggaatggg ggatgtgagt
     3061 gagaagcacg ggagtgggcc cgcagttcca gaaaaggccg ttcgtttctc tttcacagtc
     3121 atgagaatta cgatagagca tggttcacag aacgtgaagg tgtttgagga acccaagccc
     3181 aattctgaac tgtgttgcaa gccgttgtgt cttatgctgg cagatgagtc tgaccatgag
     3241 acccttactg ctattctaag ccccctcatt gccgagaggg aggccatgaa gagcagtgaa
     3301 ttaacgctgg agatgggagg catccccagg acttttaaat tcatcttcag gggcactgga
     3361 tacgatgaaa aacttgtccg ggaagtagaa ggcttggaag cttctggctc agtctacatc
     3421 tgtacactct gtgacaccac ccgtttggaa gcctctcaga atcttgtctt ccactccata
     3481 accagaagcc acgccgagaa cctgcagcgc tatgaggtct ggcggtccaa tccgtatcat
     3541 gagtccgtgg aagagctccg ggaccgggtg aaaggggtct ctgccaaacc tttcatcgag
     3601 acagtccctt ccatagatgc gcttcactgt gacattggca atgcagctga attctataag
     3661 attttccagc tggagatagg ggaagtgtat aaacatccca atgcctctaa agaggaaagg
     3721 aagagatggc aggccacgct ggacaaacat ctccggaaaa ggatgaactt aaaaccaatc
     3781 atgaggatga atggcaactt tgcccggaag cttatgaccc aagagactgt agacgcagtt
     3841 tgtgagttaa ttccttctga ggagaggcat gaagctctca gggagctcat ggacctttac
     3901 ctgaagatga aacccgtgtg gcgctcttca tgtcccgcta aagagtgtcc agagtccctc
     3961 tgtcagtaca gtttcaactc acagcgtttc gcggaactcc tctccaccaa gttcaaatat
     4021 agatacgagg gcaaaatcac caattacttt cacaaaacct tggcacatgt ccctgaaatt
     4081 attgaaaggg atggctctat cggggcctgg gcaagtgagg gaaatgaatc gggtaacaag
     4141 ctgtttagac ggtttcggaa aatgaatgcc aggcagtcca agtgctatga gatggaagat
     4201 gtcctgaaac atcactggct gtatacttca aaatacctcc agaagtttat gaatgctcat
     4261 aacgcgtaac tcgagactag tatcgcgata attcactcct caggtgcagg ctgcctatca
     4321 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc
     4381 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     4441 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     4501 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     4561 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
     4621 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     4681 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     4741 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     4801 tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc
     4861 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4921 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4981 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatcgatccg ctgcattaat
     5041 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5101 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5161 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5221 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5281 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5341 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5401 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5461 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5521 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5581 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5641 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5701 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5761 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5821 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5881 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5941 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6001 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6061 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6121 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6181 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6241 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6301 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6361 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6421 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6481 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6541 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6601 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6661 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6721 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6781 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6841 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6901 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6961 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt
     7021 cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     7081 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     7141 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     7201 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     7261 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     7321 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     7381 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
     7441 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
     7501 atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
     7561 ggcggggcga ggcggagagg tgcggcggca gcc
//
