LOCUS       sequence-192189-        6890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Flag-tagged human CELF2 (aka CUGBP2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6890)
  AUTHORS   Orengo JP, Bundman D, Cooper TA
  TITLE     A bichromatic fluorescent reporter for cell-based screens of 
            alternative splicing.
  JOURNAL   Nucleic Acids Res. 2006;34(22):e148. Epub 2006 Nov 16.
  PUBMED    17142220
REFERENCE   2  (bases 1 to 6890)
  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"; journalName: "Nucleic 
            Acids Res."; date: "2006"; volume: "34(22)"; pages: "e148. Epub 2006
            Nov 16"
COMMENT     Sequence Label: FlagETR3 (CELF2)
FEATURES             Location/Qualifiers
     source          1..6890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             922..945
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2481..2498)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2487..2711
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2757..3185
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2844..2863)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3054..3075
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3194..3214)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3199..3528
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3379..3514
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3441..3460
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3595..4389
                     /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(3649..3668)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4259..4278
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4563..4684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4600..4619)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4654..4673
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4733..4749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4733..4749)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4746..4768)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4757..4773
                     /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(4781..4787,4788..4805,4806..4811))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4826..4847
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4964..4981)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5135..5723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5215..5234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5894..6685,6686..6754))
                     /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     6517..6536
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6755..6859)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttccat ggattacaag gatgacgatg acaagggggt accaggatct
      961 atgaacggag ctttggatca ctcagaccaa ccagacccag atgccattaa gatgtttgtc
     1021 ggacagatcc cccggtcatg gtcggaaaag gagctgaaag aactttttga gccttacgga
     1081 gccgtctacc agatcaacgt cctccgggac cggagtcaga accctccgca gagtaaaggt
     1141 tgttgtttcg taacatttta tacaagaaaa gctgcacttg aggcccagaa tgcactgcac
     1201 aatattaaaa ctttacctgg gatgcatcat cccattcaga tgaaacctgc agatagtgaa
     1261 aagtccaacg ctgtggaaga cagaaaattg ttcataggaa tggtttcgaa gaaatgtaat
     1321 gagaacgaca tcagggtgat gttctctcca tttggccaga tagaagaatg ccggatcctc
     1381 cggggacctg atgggctgag tcgaggctgt gcgtttgtca cattttctac aagggcaatg
     1441 gcacagaatg caatcaaagc catgcatcag tctcagacca tggagggctg ctcttcacct
     1501 atcgtggtga agtttgctga cactcagaag gacaaagagc aaaggcgcct ccagcagcag
     1561 ctcgctcagc agatgcagca gctcaacact gccacctggg ggaacctgac agggctgggc
     1621 ggactgaccc cacagtatct ggcgctcctg cagcaggcca cctcctccag caacctgggt
     1681 gcgttcagcg gcattcaaca aatggcaggc atgaatgctt tacagttgca gaacctggcg
     1741 acgctggctg ctgctgcagc tgcggcccag acctcagcca ccagcaccaa tgcaaaccct
     1801 ctctctacca cgagcagcgc cctgggagcc ctcacgagtc ccgtggctgc ttcaaccccc
     1861 aactccactg ctggtgcagc catgaactcc ttgacctctc tcgggactct gcaaggactg
     1921 gctggagcca ctgttggact gaataatatt aatgcactag cagttgctca aatgctctca
     1981 ggtatggcgg ctctgaatgg aggacttggc gccacaggct tgacgaatgg cacggctggc
     2041 accatggacg ccctcaccca ggcctactca ggaattcaac agtacgcagc cgccgcgctg
     2101 cccactctgt acagccagag cctgctgcag cagcagagcg ctgcaggcag ccagaaggaa
     2161 ggtccagagg gggcaaacct ctttatttac caccttccac aggaatttgg agaccaggac
     2221 attctgcaga tgttcatgcc ttttggaaat gttatctctg ctaaagtctt cattgacaaa
     2281 cagaccaatc tgagcaagtg ctttggtttt gttagctacg acaatccagt ctctgcacaa
     2341 gctgctatcc aagctatgaa tggctttcag atcggcatga aacgcttgaa ggtgcagctg
     2401 aagcgttcca aaaacgacag caaaccttac tgaaagcttg accgtcgagt ctagagggcc
     2461 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2521 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2581 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2641 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     2701 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     2761 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2821 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2881 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2941 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3001 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3061 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3121 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3181 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3241 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3301 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3361 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3421 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3481 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3541 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3601 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3661 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3721 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3781 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3841 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3901 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3961 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4021 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4081 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4141 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4201 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4261 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4321 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4381 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4441 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4501 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4561 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4621 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4681 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4741 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4801 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4861 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4921 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4981 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5041 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5101 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5161 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5221 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5281 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5341 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5401 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5461 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5521 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5581 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5641 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5701 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5761 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5821 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5881 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5941 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6001 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6061 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6121 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6181 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6241 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6301 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6361 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6421 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6481 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6541 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6601 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6661 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6721 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6781 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6841 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
