LOCUS       sequence-249674-        6222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro translation of human ELL2 with Met1 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 6222)
  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 6222)
  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: Met1ILeu ELL2
FEATURES             Location/Qualifiers
     source          1..6222
                     /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(3361..3379)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3400..3416)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3400..3416)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3413..3435)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3424..3440
                     /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(3448..3454,3455..3472,3473..3478))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3493..3514
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3631..3648)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3802..4390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3882..3901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4561..5352,5353..5421))
                     /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     5184..5203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5422..5526)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5552..6007)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5689..5710)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5901..5920
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6134..6156
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6148..6165
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6149..6165
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6172..6191
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6172..6190
                     /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 cgcggcgggg 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 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 tggaaaaaaa
     2401 aaaaaaaaaa aaaaaaaaaa cgcgtaaccc cccattgcaa tggttgcctt aaggtgtttg
     2461 ctagttgtgt acatagtgtg gttaatcatt agctacactg cttcccactt gattagagca
     2521 atgggaagca tactgtggcc taccagcatc tggaagtgtg tgctcgatct gtatgtgtgc
     2581 agaggtggtg tggatgtgag cgtgcatgaa ggaaaaaaag ctgctactcc tagtaggcca
     2641 aacgctcagg ttaaacaact gacgagtgtt actgtagggt gtttttttgt ttttttgttt
     2701 tttttcttct atcaaattgc tacttttgtt gtggaagaca aaagcatttc catttcaacg
     2761 agtttgtcag ctttattaat gttgggcaaa aattgatatg tcatgaaaat gaaacagatc
     2821 tatagttttg ggacaaaatt ataaaatgaa atgtgtaggt aacctattta tatactgcta
     2881 taaagtattt tttgaagaga gatatgcaaa gaagctatta cctacataag aggtatattt
     2941 aaagattttt tttttttcat cctggtgcca ggaatataaa aaagagtgga tatatttaac
     3001 cataacatac tgtgattcat caaacagcac aaactttcat ttcatggagt ttatctgttg
     3061 acattgattt aaactgtcac ttgttttatc atgtgggaac ataagttatg tggtcaaaaa
     3121 tataaggatt ttgaattaat gttgattcaa gttgtattgt cttttcaaag tgctgccagt
     3181 tgaaaaggga agcattatgt ttacaaatct gttttgaaat gtttgccaaa attttggtag
     3241 tgtctttaat aaagatgttt gtctccagca tccagaaaaa aaaaaaaaaa actctccagc
     3301 gctggatccg gccataaggg cctgatcctt cgaggggggg cccggtacca gcttttgttc
     3361 cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3421 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3481 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3541 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3601 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3661 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3721 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3781 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3841 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3901 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3961 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4021 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4081 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4141 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4201 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4261 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4321 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4381 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4441 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4501 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4561 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4621 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4681 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4741 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4801 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4861 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4921 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4981 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5041 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5101 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5161 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5221 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5281 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5341 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5401 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5461 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5521 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     5581 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5641 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5701 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5761 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5821 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5881 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5941 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6001 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     6061 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     6121 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac
     6181 tcactatagg gcgaattgga gctccaccgc ggtggcggcc gc
//
