LOCUS       sequence-249719-        6225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro translation of human ELL2 with Met133 mutated to Ileu 
            to ask if there is internal initiation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6225)
  AUTHORS   Martincic K, Alkan SA, Cheatle A, Borghesi L, Milcarek C
  TITLE     Transcription elongation factor ELL2 directs immunoglobulin 
            secretion in plasma cells by stimulating altered RNA processing.
  JOURNAL   Nat Immunol. 2009 Oct;10(10):1102-9. doi: 10.1038/ni.1786. Epub 2009
            Sep 13.
  PUBMED    19749764
REFERENCE   2  (bases 1 to 6225)
  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: "10.1038/ni.1786"; 
            journalName: "Nat Immunol"; date: "2009-10"; volume: "10"; issue: 
            "10"; pages: "1102-9"
COMMENT     Sequence Label: Met133ILeu ELL2
FEATURES             Location/Qualifiers
     source          1..6225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(52..85)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(641..652)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(3415..3433)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3454..3470)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3454..3470)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3467..3489)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3478..3494
                     /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(3502..3508,3509..3526,3527..3532))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3547..3568
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3685..3702)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3856..4444)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3936..3955)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4615..5406,5407..5475))
                     /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     5238..5257
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5476..5580)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5606..6061)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5743..5764)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5955..5974
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6188..6210
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6202..6219
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6203..6219
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattggag ctccaccgcg gtggcggccg cataacttcg
       61 tatagcatac attatacgaa gttatggatc aggccaaatc ggccgagctc gaattcgtcg
      121 agagcgggat aagttggcgc gtcacctaga cgtcagcctc ttacctctcg gacgtcccgt
      181 cccaggagca aaactctagt gactttgggc ccgcccccgt gagagtggag ggcggccgat
      241 gagggcggtt ctgaagggaa caagaccaat gagacgagcc cggggcgggg cgcggggccg
      301 gagggtgggg cgggtggcgg cggtggcagt ccgaggtctg gctccgcgca gtatatgaca
      361 gtacgtcagc agcgggatgg ccctagcagt ggcggcggct gcagaagccc aagcagccgc
      421 ggccgcagtg gaggctagag ccggagcggc ggcggcggcg gcaccccggg gagtttaaga
      481 tggcggcggg ggggacaggg ggcctgcggg aggagcagcg ctatgggctg tcgtgcggac
      541 ggctggggca ggacaacatc accgtactgc atgtgaagct caccgagacg gcgatccggg
      601 cgctcgagac ttaccagagc cacaagaatt taattccttt tcgaccttca atccagttcc
      661 aaggactcca cgggcttgtc aaaattccca aaaatgatcc cctcaatgaa gttcataact
      721 ttaactttta tttgtcaaat gtgggcaaag acaaccctca gggcagcttt gactgcatcc
      781 agcaaacatt ctccagctct ggagcctccc agctcaattg cctgggattt atacaagata
      841 aaattacagt gtgtgcaaca aacgactcgt atcagatcac acgagaaaga atgacccagg
      901 cagaggagga atcccgcaac cgaagcacaa aagttatcaa acccggtgga ccatatgtag
      961 ggaaaagagt gcaaattcgg aaagcacctc aagctgtttc agatacagtt cctgagagga
     1021 aaaggtcaac ccccatgaac cctgcaaata caattcgaaa gacacatagc agcagcacca
     1081 tctctcagag gccatacagg gacagggtga ttcacttact ggccctgaag gcctacaaga
     1141 aaccggagct acttgctaga ctccagaaag atggtgtcaa tcaaaaagac aagaactccc
     1201 tgggagcaat tctgcaacag gtagccaatc tgaattctaa ggacctctca tataccttaa
     1261 aggattatgt ttttaaagag cttcaaagag actggcctgg atacagtgaa atagacagac
     1321 ggtcattgga gtcagtgctc tctagaaaac taaatccgtc tcagaatgct acaggcacca
     1381 gccgttcaga atctcctgta tgttctagta gagatgctgt atcttctcct cagaaacggc
     1441 ttttggattc agagtttatt gatcctttaa tgaataaaaa agcccgaata tctcacctga
     1501 cgaacagagt accaccaaca ctaaatggtc atttgaatcc caccagtgaa aaatcggctg
     1561 caggcctccc actgccccct gcggctgctg ccatccccac ccctccaccg ctgccttcaa
     1621 cctatctgcc catctcacat cctcctcaga ttgtaaattc taactccaac tcccctagca
     1681 ctccagaagg ccgggggact caagacctac ctgttgacag ttttagtcaa aacgatagta
     1741 tctatgagga ccagcaagac aaatatacct ctaggacttc tctggaaacc ttaccccctg
     1801 gttccgttct actaaagtgt ccaaagccta tggaagaaaa ccattcaatg tctcacaaaa
     1861 agtccaaaaa gaagtctaaa aaacataagg aaaaggacca aataaaaaag cacgacattg
     1921 agactattga ggaaaaggag gaagatctta agagagaaga ggaaattgcc aagctaaata
     1981 actccagtcc aaattccagt ggaggagtta aagaggattg cactgcctcc atggaacctt
     2041 cagcaattga actcccagat tatttgataa aatatatcgc tatcgtctcc tatgagcaac
     2101 gccagaatta taaggatgac ttcaatgcag agtatgatga gtacagagct ttgcatgcca
     2161 ggatggagac tgtagctaga agatttatca aactagatgc acaaagaaag cgcctttctc
     2221 caggctcaaa agagtatcag aatgttcatg aagaagtctt acaagaatat cagaagataa
     2281 agcagtctag tcccaattac catgaagaaa aatacagatg tgaatatctt cataacaagc
     2341 tggctcacat caaaaggcta ataggtgaat ttgaccaaca gcaagcagag tcatggtcct
     2401 agaactctgc ttggaccaga agatgtgaat aaacttaagc ttggaaaaaa aaaaaaaaaa
     2461 aaaaaaaaaa aaacgcgtaa ccccccattg caatggttgc cttaaggtgt ttgctagttg
     2521 tgtacatagt gtggttaatc attagctaca ctgcttccca cttgattaga gcaatgggaa
     2581 gcatactgtg gcctaccagc atctggaagt gtgtgctcga tctgtatgtg tgcagaggtg
     2641 gtgtggatgt gagcgtgcat gaaggaaaaa aagctgctac tcctagtagg ccaaacgctc
     2701 aggttaaaca actgacgagt gttactgtag ggtgtttttt tgtttttttg ttttttttct
     2761 tctatcaaat tgctactttt gttgtggaag acaaaagcat ttccatttca acgagtttgt
     2821 cagctttatt aatgttgggc aaaaattgat atgtcatgaa aatgaaacag atctatagtt
     2881 ttgggacaaa attataaaat gaaatgtgta ggtaacctat ttatatactg ctataaagta
     2941 ttttttgaag agagatatgc aaagaagcta ttacctacat aagaggtata tttaaagatt
     3001 tttttttttt catcctggtg ccaggaatat aaaaaagagt ggatatattt aaccataaca
     3061 tactgtgatt catcaaacag cacaaacttt catttcatgg agtttatctg ttgacattga
     3121 tttaaactgt cacttgtttt atcatgtggg aacataagtt atgtggtcaa aaatataagg
     3181 attttgaatt aatgttgatt caagttgtat tgtcttttca aagtgctgcc agttgaaaag
     3241 ggaagcatta tgtttacaaa tctgttttga aatgtttgcc aaaattttgg tagtgtcttt
     3301 aataaagatg tttgtctcca gcatccagna aaaaaaaaaa aaaaactctc cagcgctgga
     3361 tccggccata agggcctgat ccttcgaggg ggggcccggt accagctttt gttcccttta
     3421 gtgagggtta atttcgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3481 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3541 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3601 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3661 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3721 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3781 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3841 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3901 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3961 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4021 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4081 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4141 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4201 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4261 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4321 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4381 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4441 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4501 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4561 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4621 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4681 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4741 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4801 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4861 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4921 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4981 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5041 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5101 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5161 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5221 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5281 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5341 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5401 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5461 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5521 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5581 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     5641 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5701 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5761 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5821 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5881 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5941 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     6001 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     6061 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     6121 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     6181 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattg
//
