LOCUS       sequence-250000-        6224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro translation of human ELL2 with Met138 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 6224)
  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 6224)
  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: Met138ILeu ELL2
FEATURES             Location/Qualifiers
     source          1..6224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..34)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(590..601)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     gap             2391..2421
                     /estimated_length=31
     promoter        complement(3363..3381)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3402..3418)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3402..3418)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3415..3437)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3426..3442
                     /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(3450..3456,3457..3474,3475..3480))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3495..3516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3633..3650)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3804..4392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3884..3903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4563..5354,5355..5423))
                     /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     5186..5205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5424..5528)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5554..6009)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5691..5712)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5903..5922
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6136..6158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6150..6167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6151..6167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6174..6193
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6174..6192
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ataacttcgt atagcataca ttatacgaag ttatggatca ggccaaatcg gccgagctcg
       61 aattcgtcga gagcgggata agttggcgcg tcacctagac gtcagcctct tacctctcgg
      121 acgtcccgtc ccaggagcaa aactctagtg actttgggcc cgcccccgtg agagtggagg
      181 gcggccgatg agggcggttc tgaagggaac aagaccaatg agacgagccc ggggcggggc
      241 gcggggccgg agggtggggc gggtggcggc ggtggcagtc cgaggtctgg ctccgcgcag
      301 tatatgacag tacgtcagca gcgggatggc cctagcagtg gcggcggctg cagaagccca
      361 agcagccgcg gccgcagtgg aggctagagc cggagcggcg gcggcggcgg caccccgggg
      421 agtttaagat ggcggcgggg gggacagggg gcctgcggga ggagcagcgc tatgggctgt
      481 cgtgcggacg gctggggcag gacaacatca ccgtactgca tgtgaagctc accgagacgg
      541 cgatccgggc gctcgagact taccagagcc acaagaattt aattcctttt cgaccttcaa
      601 tccagttcca aggactccac gggcttgtca aaattcccaa aaatgatccc ctcaatgaag
      661 ttcataactt taacttttat ttgtcaaatg tgggcaaaga caaccctcag ggcagctttg
      721 actgcatcca gcaaacattc tccagctctg gagcctccca gctcaattgc ctgggattta
      781 tacaagataa aattacagtg tgtgcaacaa acgactcgta tcagatgaca cgagaaagaa
      841 tcacccaggc agaggaggaa tcccgcaacc gaagcacaaa agttatcaaa cccggtggac
      901 catatgtagg gaaaagagtg caaattcgga aagcacctca agctgtttca gatacagttc
      961 ctgagaggaa aaggtcaacc cccatgaacc ctgcaaatac aattcgaaag acacatagca
     1021 gcagcaccat ctctcagagg ccatacaggg acagggtgat tcacttactg gccctgaagg
     1081 cctacaagaa accggagcta cttgctagac tccagaaaga tggtgtcaat caaaaagaca
     1141 agaactccct gggagcaatt ctgcaacagg tagccaatct gaattctaag gacctctcat
     1201 ataccttaaa ggattatgtt tttaaagagc ttcaaagaga ctggcctgga tacagtgaaa
     1261 tagacagacg gtcattggag tcagtgctct ctagaaaact aaatccgtct cagaatgcta
     1321 caggcaccag ccgttcagaa tctcctgtat gttctagtag agatgctgta tcttctcctc
     1381 agaaacggct tttggattca gagtttattg atcctttaat gaataaaaaa gcccgaatat
     1441 ctcacctgac gaacagagta ccaccaacac taaatggtca tttgaatccc accagtgaaa
     1501 aatcggctgc aggcctccca ctgccccctg cggctgctgc catccccacc cctccaccgc
     1561 tgccttcaac ctatctgccc atctcacatc ctcctcagat tgtaaattct aactccaact
     1621 cccctagcac tccagaaggc cgggggactc aagacctacc tgttgacagt tttagtcaaa
     1681 acgatagtat ctatgaggac cagcaagaca aatatacctc taggacttct ctggaaacct
     1741 taccccctgg ttccgttcta ctaaagtgtc caaagcctat ggaagaaaac cattcaatgt
     1801 ctcacaaaaa gtccaaaaag aagtctaaaa aacataagga aaaggaccaa ataaaaaagc
     1861 acgacattga gactattgag gaaaaggagg aagatcttaa gagagaagag gaaattgcca
     1921 agctaaataa ctccagtcca aattccagtg gaggagttaa agaggattgc actgcctcca
     1981 tggaaccttc agcaattgaa ctcccagatt atttgataaa atatatcgct atcgtctcct
     2041 atgagcaacg ccagaattat aaggatgact tcaatgcaga gtatgatgag tacagagctt
     2101 tgcatgccag gatggagact gtagctagaa gatttatcaa actagatgca caaagaaagc
     2161 gcctttctcc aggctcaaaa gagtatcaga atgttcatga agaagtctta caagaatatc
     2221 agaagataaa gcagtctagt cccaattacc atgaagaaaa atacagatgt gaatatcttc
     2281 ataacaagct ggctcacatc aaaaggctaa taggtgaatt tgaccaacag caagcagagt
     2341 catggtccta gaactctgct tggaccagaa gatgtgaata aacttaagct nnnnnnnnnn
     2401 nnnnnnnnnn nnnnnnnnnn nacgcgtaac cccccattgc aatggttgcc ttaaggtgtt
     2461 tgctagttgt gtacatagtg tggttaatca ttagctacac tgcttcccac ttgattagag
     2521 caatgggaag catactgtgg cctaccagca tctggaagtg tgtgctcgat ctgtatgtgt
     2581 gcagaggtgg tgtggatgtg agcgtgcatg aaggaaaaaa agctgctact cctagtaggc
     2641 caaacgctca ggttaaacaa ctgacgagtg ttactgtagg gtgttttttt gtttttttgt
     2701 tttttttctt ctatcaaatt gctacttttg ttgtggaaga caaaagcatt tccatttcaa
     2761 cgagtttgtc agctttatta atgttgggca aaaattgata tgtcatgaaa atgaaacaga
     2821 tctatagttt tgggacaaaa ttataaaatg aaatgtgtag gtaacctatt tatatactgc
     2881 tataaagtat tttttgaaga gagatatgca aagaagctat tacctacata agaggtatat
     2941 ttaaagattt tttttttttc atcctggtgc caggaatata aaaaagagtg gatatattta
     3001 accataacat actgtgattc atcaaacagc acaaactttc atttcatgga gtttatctgt
     3061 tgacattgat ttaaactgtc acttgtttta tcatgtggga acataagtta tgtggtcaaa
     3121 aatataagga ttttgaatta atgttgattc aagttgtatt gtcttttcaa agtgctgcca
     3181 gttgaaaagg gaagcattat gtttacaaat ctgttttgaa atgtttgcca aaattttggt
     3241 agtgtcttta ataaagatgt ttgtctccag catccagaaa aaaaaaaaaa aaactctcca
     3301 gcgctggatc cggccataag ggcctgatcc ttcgaggggg ggcccggtac cagcttttgt
     3361 tccctttagt gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg
     3421 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3481 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3541 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3601 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3661 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3721 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3781 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3841 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3901 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3961 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4021 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4081 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4141 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4201 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4261 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4321 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4381 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4441 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4501 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4561 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4621 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4681 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4741 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4801 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4861 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4921 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4981 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5041 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5101 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5161 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5221 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5281 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5341 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5401 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5461 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5521 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     5581 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     5641 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     5701 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     5761 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     5821 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     5881 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     5941 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     6001 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     6061 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     6121 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg
     6181 actcactata gggcgaattg gagctccacc gcggtggcgg ccgc
//
