LOCUS       sequence-250013-        6299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro translation of human ELL2 with Met133, 138, 186 to Ileu
            with HA tag after 8th Met.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6299)
  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 6299)
  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: HA tagged Sph ELL2 Met133, 138, 186 to Ileu
FEATURES             Location/Qualifiers
     source          1..6299
                     /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"
     CDS             2109..2135
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(2146..2172)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3438..3456)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3477..3493)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3477..3493)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3490..3512)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3501..3517
                     /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(3525..3531,3532..3549,3550..3555))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3570..3591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3708..3725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3879..4467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3959..3978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4638..5429,5430..5498))
                     /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     5261..5280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5499..5603)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5629..6084)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5766..5787)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5978..5997
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6211..6233
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6225..6242
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6226..6242
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6249..6268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6249..6267
                     /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 tcagatcaca cgagaaagaa
      841 tcacccaggc 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 tgcatgggta cccatacgat gtcccagatt acgctgctca tgagcagcgt agtctgggac
     2161 gtcgtatggg tacccatgcc aggatggaga ctgtagctag aagatttatc aaactagatg
     2221 cacaaagaaa gcgcctttct ccaggctcaa aagagtatca gaatgttcat gaagaagtct
     2281 tacaagaata tcagaagata aagcagtcta gtcccaatta ccatgaagaa aaatacagat
     2341 gtgaatatct tcataacaag ctggctcaca tcaaaaggct aataggtgaa tttgaccaac
     2401 agcaagcaga gtcatggtcc tagaactctg cttggaccag aagatgtgaa taaacttaag
     2461 ctatcmarma marwaaaaaa aaaaaaaaaa aaraarcgcg taacccccca ttgcaatggt
     2521 tgccttaagg tgtttgctag ttgtgtacat agtgtggtta atcattagct acactgcttc
     2581 ccacttgatt agagcaatgg gaagcatact gtggcctacc agcatctgga agtgtgtgct
     2641 cgatctgtat gtgtgcagag gtggtgtgga tgtgagcgtg catgaaggaa aaaaagctgc
     2701 tactcctagt aggccaaacg ctcaggttaa acaactgacg agtgttactg tagggtgttt
     2761 ttttgttttt ttgttttttt tcttctatca aattgctact tttgttgtgg aagacaaaag
     2821 catttccatt tcaacgagtt tgtcagcttt attaatgttg ggcaaaaatt gatatgtcat
     2881 gaaaatgaaa cagatctata gttttgggac aaaattataa aatgaaatgt gtaggtaacc
     2941 tatttatata ctgctataaa gtattttttg aagagagata tgcaaagaag ctattaccta
     3001 cataagaggt atatttaaag attttttttt ttttcatcct ggtgccagga atataaaaaa
     3061 gagtggatat atttaaccat aacatactgt gattcatcaa acagcacaaa ctttcatttc
     3121 atggagttta tctgttgaca ttgatttaaa ctgtcacttg ttttatcatg tgggaacata
     3181 agttatgtgg tcaaaaatat aaggattttg aattaatgtt gattcaagtt gtattgtctt
     3241 ttcaaagtgc tgccagttga aaagggaagc attatgttta caaatctgtt ttgaaatgtt
     3301 tgccaaaatt ttggtagtgt ctttaataaa gatgtttgtc tccagcatcc agaaaaaaaa
     3361 aaaaaaaact ctccagcgct ggatccggcc ataagggcct gatccttcga gggggggccc
     3421 ggtaccagct tttgttccct ttagtgaggg ttaatttcga gcttggcgta atcatggtca
     3481 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3541 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3601 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3661 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3721 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3781 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3841 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3901 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3961 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4021 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4081 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4141 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4201 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4261 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4321 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4381 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4441 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4501 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4561 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4621 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4681 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4741 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4801 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4861 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4921 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4981 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5041 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5101 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5161 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5221 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5281 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5341 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5401 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5461 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5521 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5581 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     5641 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     5701 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     5761 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     5821 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     5881 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     5941 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6001 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6061 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     6121 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     6181 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     6241 gtgaattgta atacgactca ctatagggcg aattggagct ccaccgcggt ggcggccgc
//
