LOCUS       sequence-249997-        6225 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For in vitro translation of human ELL2 with Met186 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 Apr 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: Met186ILeu ELL2
FEATURES             Location/Qualifiers
     source          1..6225
                     /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"
     promoter        complement(3364..3382)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3403..3419)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3403..3419)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3416..3438)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3427..3443
                     /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(3451..3457,3458..3475,3476..3481))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3496..3517
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3634..3651)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3805..4393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3885..3904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4564..5355,5356..5424))
                     /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     5187..5206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5425..5529)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5555..6010)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5692..5713)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5904..5923
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6137..6159
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6151..6168
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6152..6168
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6175..6194
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6175..6193
                     /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 tgacccaggc agaggaggaa tcccgcaacc gaagcacaaa agttatcaaa cccggtggac
      901 catatgtagg gaaaagagtg caaattcgga aagcacctca agctgtttca gatacagttc
      961 ctgagaggaa aaggtcaacc cccatcaacc 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 tggaaaaaaa
     2401 aaaaaaaaaa aaaaaaaaaa aaacgcgtaa ccccccattg caatggttgc cttaaggtgt
     2461 ttgctagttg tgtacatagt gtggttaatc attagctaca ctgcttccca cttgattaga
     2521 gcaatgggaa gcatactgtg gcctaccagc atctggaagt gtgtgctcga tctgtatgtg
     2581 tgcagaggtg gtgtggatgt gagcgtgcat gaaggaaaaa aagctgctac tcctagtagg
     2641 ccaaacgctc aggttaaaca actgacgagt gttactgtag ggtgtttttt tgtttttttg
     2701 ttttttttct tctatcaaat tgctactttt gttgtggaag acaaaagcat ttccatttca
     2761 acgagtttgt cagctttatt aatgttgggc aaaaattgat atgtcatgaa aatgaaacag
     2821 atctatagtt ttgggacaaa attataaaat gaaatgtgta ggtaacctat ttatatactg
     2881 ctataaagta ttttttgaag agagatatgc aaagaagcta ttacctacat aagaggtata
     2941 tttaaagatt tttttttttt catcctggtg ccaggaatat aaaaaagagt ggatatattt
     3001 aaccataaca tactgtgatt catcaaacag cacaaacttt catttcatgg agtttatctg
     3061 ttgacattga tttaaactgt cacttgtttt atcatgtggg aacataagtt atgtggtcaa
     3121 aaatataagg attttgaatt aatgttgatt caagttgtat tgtcttttca aagtgctgcc
     3181 agttgaaaag ggaagcatta tgtttacaaa tctgttttga aatgtttgcc aaaattttgg
     3241 tagtgtcttt aataaagatg tttgtctcca gcatccagaa aaaaaaaaaa aaaactctcc
     3301 agcgctggat ccggccataa gggcctgatc cttcgagggg gggcccggta ccagcttttg
     3361 ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt
     3421 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3481 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3541 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3601 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3661 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3721 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3781 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3841 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3901 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3961 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4021 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4081 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4141 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4201 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4261 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4321 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4381 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4441 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4501 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4561 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4621 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4681 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4741 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4801 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4861 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4921 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4981 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5041 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5101 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5161 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5221 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5281 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5341 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5401 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5461 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5521 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     5581 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     5641 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     5701 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     5761 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     5821 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     5881 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     5941 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     6001 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     6061 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     6121 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
     6181 gactcactat agggcgaatt ggagctccac cgcggtggcg gccgc
//
