LOCUS       sequence-250029-        7681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Dominant Negative CstF64 in mammalian cells; DN blocks
            polyadenylation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7681)
  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 7681)
  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: Dom neg CstF64
FEATURES             Location/Qualifiers
     source          1..7681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1196..1215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1196..1214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1257..1273
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1258..1274
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2795..2821
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2846..2875
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2891..2908
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2931..2948)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2937..3161
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3207..3635
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3294..3313)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3504..3525
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3644..3664)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3649..3978
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3829..3964
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3891..3910
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4045..4839
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4099..4118)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4709..4728
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5015..5136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5052..5071)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5106..5125
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5185..5201)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5185..5201)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5198..5220)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5209..5225
                     /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(5233..5239,5240..5257,5258..5263))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5278..5299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5416..5433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5576..6164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5656..5675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6335..7126,7127..7195))
                     /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     6958..6977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7196..7300)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7375..7394)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgggc cccccctcga
     1261 ggtcgacggt atcgataagc ttgatatcga attccggaat tccggaattc ccggaagccg
     1321 actcaacaga gctatggcgg gtttgactgt gagagaccca gcggtggatc gttctctacg
     1381 ttctgtgttc gtggggaaca ttccttatga agctactgaa gagcagttga aggacatctt
     1441 ttctgaggtt ggacctgttg ttagtttcag attggtatac gatagagaga caggaaagcc
     1501 aaagggttat ggcttctgtg aataccaaga ccaagagaca gcacttagtg ccatgcggaa
     1561 cctgaatggg cgcgaattca gtgggagagc acttcgagtg gacaatgctg ccagtgaaaa
     1621 gaacaaagaa gagctgaaga gccttggcac tggtgcccct gtcattgagt caccttatgg
     1681 agagaccatc agtcctgagg atgcccctga gtccattagc aaagcagttg ccagccttcc
     1741 accagagcag atgtttgagc tgatgaaaca aatgaagctc tgtgtccaga atagtcccca
     1801 ggaggcacgg aacatgttac ttcagaaccc tcaactggct tatgctttgc tgcaagcaca
     1861 ggtagtgatg agaattgtgg atccggaaat tgccctgaaa attctgcatc gccagacaaa
     1921 tatcccaacg ctgattgcag gcaaccctca gccagtccat ggtgctgggc ctggctcagg
     1981 atccaatgtg tcaatgaacc agcagaatcc tcaggcccct caggcccagt ctttgggtgg
     2041 aatgcatgtc aatggcgcac ctcctctgat gcaagcttct atgcagggtg gagttccagc
     2101 accagggcaa atgccagctg ctgtcacagg acctggccct ggttccttag ctcctggagg
     2161 aggaatgcag gctcaggttg gaatgccagg aagtggacca gtgtccatgg aacgggggca
     2221 agtgccgatg caagacccca gagcagctat gcagcgggga tccttgcctg cgaatgtccc
     2281 aacccctcga ggcttgttag gagatgctcc gaatgatcca cggggaggca ctttactttc
     2341 tgtaactgga gaggtagagc ctagaggtta cttgggacca cctcatcagg gtccacccat
     2401 gcaccatgtc cctggccatg agagccgagg accaccccca catgaactga ggggagggcc
     2461 attacccgag cccagacctc taatggcaga accaagagga cccatgctag atcagagggg
     2521 tccacccttg gatggcagag gtggaaggga tccccgagga atagatgcac gagggatgga
     2581 ggcccgagcc atggaggcaa gagggttaga tgccagagga ttagaggccc gtgcaatgga
     2641 ggcccgtgcg atggaagctc gtgcaatgga ggcccgagcg atggaggccc gtgcaatgga
     2701 agtccgaggg atggaggcca gaggcatgga taccagaggc ccagtgcctg gccccagagg
     2761 acctatacct agtggaatgc agggtccagg cgcctaccca tacgacgtcc cagactacgc
     2821 tgctagtcta gagggcccgc ggttcgaaca aaaactcatc tcagaagagg atctgaatat
     2881 gcataccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     2941 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3001 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3061 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3121 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3181 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3241 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3301 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3361 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3421 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3481 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3541 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3601 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3661 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3721 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3781 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3841 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3901 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3961 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4021 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4081 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4141 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4201 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4261 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4321 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4381 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4441 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4501 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4561 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4621 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4681 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4741 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4801 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     4861 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4921 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4981 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5041 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5101 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5161 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5221 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5281 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5341 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5401 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5461 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5521 aaagaacatg tgagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5581 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5641 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5701 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5761 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5821 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5881 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5941 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6001 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6061 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6121 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6181 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6241 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6301 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6361 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6421 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6481 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6541 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6601 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6661 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6721 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6781 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6841 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6901 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6961 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7021 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7081 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7141 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7201 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7261 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7321 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     7381 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     7441 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     7501 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     7561 gcgttgacat tgattattga ctaggctttt gcaaaaagct ttgcaaagat ggataaagtt
     7621 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
     7681 t
//
