LOCUS       sequence-250039-        7782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ELL2 with M133, 138, 186ILeu to prevent internal Met
            initiation peptides and contains HA tag near C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7782)
  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 7782)
  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 SphI ELL2 M133, 138, 186ILeu
FEATURES             Location/Qualifiers
     source          1..7782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..36
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        17..35
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             77..94
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             98..130
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(134..142,143..157)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(367..378)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1886..1912
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2261..2278)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2267..2491
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2537..2965
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2624..2643)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2834..2855
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2974..2994)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2979..3308
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3159..3294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3221..3240
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3356..3403
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3422..3796
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3926..4047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3963..3982)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4017..4036
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4096..4112)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4096..4112)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4109..4131)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4120..4136
                     /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(4144..4150,4151..4168,4169..4174))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4189..4210
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4327..4344)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4498..5086)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4578..4597)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5257..6048,6049..6117))
                     /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     5880..5899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6118..6222)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6297..6316)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6604..7782
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6835..7773
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7730..7750
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 ggaattagct tggtactaat acgactcact atagggagac ccaagctggc taggtaagct
       61 taccatgggg ggttctcatc atcatcatca tcatggtatg gctagcatga ctggtggaca
      121 gcaaatgggt cgggatctgt acgacgatga cgataaggta cctaaggatc cagtgtggtg
      181 gaattctgca gatggggagt ttaagatggc ggcggggggg acagggggcc tgcgggagga
      241 gcagcgctat gggctgtcgt gcggacggct ggggcaggac aacatcaccg tactgcatgt
      301 gaagctcacc gagacggcga tccgggcgct cgagacttac cagagccaca agaatttaat
      361 tccttttcga ccttcaatcc agttccaagg actccacggg cttgtcaaaa ttcccaaaaa
      421 tgatcccctc aatgaagttc ataactttaa cttttatttg tcaaatgtgg gcaaagacaa
      481 ccctcagggc agctttgact gcatccagca aacattctcc agctctggag cctcccagct
      541 caattgcctg ggatttatac aagataaaat tacagtgtgt gcaacaaacg actcgtatca
      601 gatcacacga gaaagaatca cccaggcaga ggaggaatcc cgcaaccgaa gcacaaaagt
      661 tatcaaaccc ggtggaccat atgtagggaa aagagtgcaa attcggaaag cacctcaagc
      721 tgtttcagat acagttcctg agaggaaaag gtcaaccccc atcaaccctg caaatacaat
      781 tcgaaagaca catagcagca gcaccatctc tcagaggcca tacagggaca gggtgattca
      841 cttactggcc ctgaaggcct acaagaaacc ggagctactt gctagactcc agaaagatgg
      901 tgtcaatcaa aaagacaaga actccctggg agcaattctg caacaggtag ccaatctgaa
      961 ttctaaggac ctctcatata ccttaaagga ttatgttttt aaagagcttc aaagagactg
     1021 gcctggatac agtgaaatag acagacggtc attggagtca gtgctctcta gaaaactaaa
     1081 tccgtctcag aatgctacag gcaccagccg ttcagaatct cctgtatgtt ctagtagaga
     1141 tgctgtatct tctcctcaga aacggctttt ggattcagag tttattgatc ctttaatgaa
     1201 taaaaaagcc cgaatatctc acctgacgaa cagagtacca ccaacactaa atggtcattt
     1261 gaatcccacc agtgaaaaat cggctgcagg cctcccactg ccccctgcgg ctgctgccat
     1321 ccccacccct ccaccgctgc cttcaaccta tctgcccatc tcacatcctc ctcagattgt
     1381 aaattctaac tccaactccc ctagcactcc agaaggccgg gggactcaag acctacctgt
     1441 tgacagtttt agtcaaaacg atagtatcta tgaggaccag caagacaaat atacctctag
     1501 gacttctctg gaaaccttac cccctggttc cgttctacta aagtgtccaa agcctatgga
     1561 agaaaaccat tcaatgtctc acaaaaagtc caaaaagaag tctaaaaaac ataaggaaaa
     1621 ggaccaaata aaaaagcacg acattgagac tattgaggaa aaggaggaag atcttaagag
     1681 agaagaggaa attgccaagc taaataactc cagtccaaat tccagtggag gagttaaaga
     1741 ggattgcact gcctccatgg aaccttcagc aattgaactc ccagattatt tgataaaata
     1801 tatcgctatc gtctcctatg agcaacgcca gaattataag gatgacttca atgcagagta
     1861 tgatgagtac agagctttgc atgggtaccc atacgacgtc ccagactacg ctgctcatgc
     1921 caggatggag actgtagcta gaagatttat caaactagat gcacaaagaa agcgcctttc
     1981 tccaggctca aaagagtatc agaatgttca tgaagaagtc ttacaagaat atcagaagat
     2041 aaagcagtct agtcccaatt accatgaaga aaaatacaga tgtgaatatc ttcataacaa
     2101 gctggctcac atcaaaaggc taataggtga atttgaccaa cagcaagcag agtcatggtc
     2161 ctagaactct gcttggacca gaagatgtga ataaacttaa gctatccagc acagtggcgg
     2221 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     2281 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2341 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2401 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2461 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2521 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2581 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2641 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2701 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2761 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2821 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2881 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2941 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3001 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3061 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3121 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3181 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3241 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3301 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     3361 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     3421 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     3481 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     3541 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     3601 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     3661 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     3721 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     3781 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     3841 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3901 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     3961 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4021 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4081 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4141 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4201 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4261 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4321 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4381 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4441 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4501 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4561 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4621 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4681 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4741 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4801 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4861 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4921 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4981 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5041 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5101 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5161 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5221 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5281 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5341 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5401 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5461 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5521 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5581 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5641 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5701 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5761 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5821 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5881 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5941 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6001 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6061 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6121 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6181 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6241 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     6301 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     6361 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     6421 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     6481 acgcgttgac attgattatt gactaggctt ttgcaaaaag ctttgcaaag atggataaag
     6541 ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa aggagtggga
     6601 attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     6661 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
     6721 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     6781 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
     6841 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
     6901 attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
     6961 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
     7021 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
     7081 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
     7141 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
     7201 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
     7261 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
     7321 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
     7381 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
     7441 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
     7501 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
     7561 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     7621 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     7681 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     7741 acagtggttc aaagtttttt tcttccattt caggtgtcgt ga
//
