LOCUS       sequence-249998-        7782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ELL2 with M133, 138, 186ILeu to prevent internal Met
            initiation peptides and contains middle HA tag that doesn't disrupt 
            function.
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 mid ELL2 M133, 138, 186ILeu
FEATURES             Location/Qualifiers
     source          1..7782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     53..73
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     122..141
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        122..140
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             182..199
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             203..235
                     /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(239..247,248..262)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(472..483)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1181..1207
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2366..2383)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2372..2596
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2642..3070
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2729..2748)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2939..2960
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3079..3099)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3084..3413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3264..3399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3326..3345
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3461..3508
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3527..3901
                     /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    4031..4152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4068..4087)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4122..4141
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4201..4217)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4201..4217)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4214..4236)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4225..4241
                     /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(4249..4255,4256..4273,4274..4279))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4294..4315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4432..4449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4603..5191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4683..4702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5362..6153,6154..6222))
                     /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     5985..6004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6223..6327)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6402..6421)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(6709..7782,1..105)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(6940..7782,1..96)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
ORIGIN
        1 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
       61 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaggaat tagcttggta
      121 ctaatacgac tcactatagg gagacccaag ctggctaggt aagcttacca tggggggttc
      181 tcatcatcat catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga
      241 tctgtacgac gatgacgata aggtacctaa ggatccagtg tggtggaatt ctgcagatgg
      301 ggagtttaag atggcggcgg gggggacagg gggcctgcgg gaggagcagc gctatgggct
      361 gtcgtgcgga cggctggggc aggacaacat caccgtactg catgtgaagc tcaccgagac
      421 ggcgatccgg gcgctcgaga cttaccagag ccacaagaat ttaattcctt ttcgaccttc
      481 aatccagttc caaggactcc acgggcttgt caaaattccc aaaaatgatc ccctcaatga
      541 agttcataac tttaactttt atttgtcaaa tgtgggcaaa gacaaccctc agggcagctt
      601 tgactgcatc cagcaaacat tctccagctc tggagcctcc cagctcaatt gcctgggatt
      661 tatacaagat aaaattacag tgtgtgcaac aaacgactcg tatcagatca cacgagaaag
      721 aatcacccag gcagaggagg aatcccgcaa ccgaagcaca aaagttatca aacccggtgg
      781 accatatgta gggaaaagag tgcaaattcg gaaagcacct caagctgttt cagatacagt
      841 tcctgagagg aaaaggtcaa cccccatcaa ccctgcaaat acaattcgaa agacacatag
      901 cagcagcacc atctctcaga ggccatacag ggacagggtg attcacttac tggccctgaa
      961 ggcctacaag aaaccggagc tacttgctag actccagaaa gatggtgtca atcaaaaaga
     1021 caagaactcc ctgggagcaa ttctgcaaca ggtagccaat ctgaattcta aggacctctc
     1081 atatacctta aaggattatg tttttaaaga gcttcaaaga gactggcctg gatacagtga
     1141 aatagacaga cggtcattgg agtcagtgct ctctaggggg tacccatacg acgtcccaga
     1201 ctacgctgct agaaaactaa atccgtctca gaatgctaca ggcaccagcc gttcagaatc
     1261 tcctgtatgt tctagtagag atgctgtatc ttctcctcag aaacggcttt tggattcaga
     1321 gtttattgat cctttaatga ataaaaaagc ccgaatatct cacctgacga acagagtacc
     1381 accaacacta aatggtcatt tgaatcccac cagtgaaaaa tcggctgcag gcctcccact
     1441 gccccctgcg gctgctgcca tccccacccc tccaccgctg ccttcaacct atctgcccat
     1501 ctcacatcct cctcagattg taaattctaa ctccaactcc cctagcactc cagaaggccg
     1561 ggggactcaa gacctacctg ttgacagttt tagtcaaaac gatagtatct atgaggacca
     1621 gcaagacaaa tatacctcta ggacttctct ggaaacctta ccccctggtt ccgttctact
     1681 aaagtgtcca aagcctatgg aagaaaacca ttcaatgtct cacaaaaagt ccaaaaagaa
     1741 gtctaaaaaa cataaggaaa aggaccaaat aaaaaagcac gacattgaga ctattgagga
     1801 aaaggaggaa gatcttaaga gagaagagga aattgccaag ctaaataact ccagtccaaa
     1861 ttccagtgga ggagttaaag aggattgcac tgcctccatg gaaccttcag caattgaact
     1921 cccagattat ttgataaaat atatcgctat cgtctcctat gagcaacgcc agaattataa
     1981 ggatgacttc aatgcagagt atgatgagta cagagctttg catgccagga tggagactgt
     2041 agctagaaga tttatcaaac tagatgcaca aagaaagcgc ctttctccag gctcaaaaga
     2101 gtatcagaat gttcatgaag aagtcttaca agaatatcag aagataaagc agtctagtcc
     2161 caattaccat gaagaaaaat acagatgtga atatcttcat aacaagctgg ctcacatcaa
     2221 aaggctaata ggtgaatttg accaacagca agcagagtca tggtcctaga actctgcttg
     2281 gaccagaaga tgtgaataaa cttaagctat ccagcacagt ggcggccgct cgagtctaga
     2341 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2401 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2461 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2521 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2581 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2641 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2701 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2761 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2821 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2881 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2941 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3001 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3061 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3241 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3301 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3361 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3421 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     3481 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac
     3541 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
     3601 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
     3661 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
     3721 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
     3781 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
     3841 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
     3901 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3961 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4021 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4081 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4141 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4201 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4261 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4321 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4381 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4441 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4501 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4561 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4621 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4681 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4741 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4801 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4861 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4921 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4981 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5041 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5101 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5161 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5221 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5281 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5341 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5401 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5461 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5521 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5581 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5641 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5701 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5761 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5821 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5881 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5941 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6001 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6061 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6121 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6181 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6241 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6301 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     6361 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     6421 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     6481 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     6541 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     6601 ttattgacta ggcttttgca aaaagctttg caaagatgga taaagtttta aacagagagg
     6661 aatctttgca gctaatggac cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc
     6721 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     6781 aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     6841 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     6901 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc
     6961 cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg
     7021 ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg
     7081 ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg
     7141 ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct
     7201 ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt
     7261 gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac
     7321 ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg
     7381 agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg
     7441 ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg
     7501 gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg
     7561 ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc
     7621 gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc
     7681 ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac
     7741 actgagtggg tggagactga agttaggcca gcttggcact tg
//
