LOCUS       sequence-434251-        8081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally EGFP-tagged IFT52 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8081)
  AUTHORS   Takei R, Katoh Y, Nakayama K
  TITLE     Robust interaction of IFT70 with IFT52-IFT88 in the IFT-B complex is
            required for ciliogenesis.
  JOURNAL   Biol Open. 2018 Apr 30;7(5). pii: bio.033241. doi: 
            10.1242/bio.033241.
  PUBMED    29654116
REFERENCE   2  (bases 1 to 8081)
  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: "Biol 
            Open."; date: "2018-04-30"; volume: "7(5). pii: bio.033241. doi"; 
            pages: " 10.1242/bio.033241"
COMMENT     Sequence Label: pCAG-EGFP-C-IFT52
FEATURES             Location/Qualifiers
     source          1..8081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1744..1753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1750..1752,1753..1755,1756..2466)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1750..1752,1753..1755,1756..2466)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1795..1816)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2056..2075)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2403..2424
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3953..3972)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4018..4073
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4072..4091)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4434..4450)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4434..4450)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4447..4469)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4458..4474
                     /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(4482..4488,4489..4506,4507..4512))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4527..4548
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4607..4802
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4653..4788
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4808..4942
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4845..4864)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4899..4918
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5010..5027)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5181..5769)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5261..5280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5940..6731,6732..6800))
                     /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     6563..6582
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6801..6905)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7217..8020
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7280..7299)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7890..7909
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8027..8075
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa tttgctagtt
     1741 ttcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1801 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1861 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1921 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     1981 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2041 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2101 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2161 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2221 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     2281 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2341 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2401 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2461 tacaagtccg gactcagatc tatggagaaa gagctgcgga gcaccattct tttcaatgcc
     2521 tacaaaaagg agatatttac caccaacaat ggctacaaat ccatgcagaa aaaacttcgg
     2581 agtaattgga agattcagag cttaaaagat gaaatcacat ctgagaagtt aaatggagta
     2641 aaactgtgga ttacagctgg gccaagggaa aaatttactg cagctgagtt tgaaatcctg
     2701 aagaaatatc ttgacactgg tggagatgtc tttgtgatgc taggagaagg tggagaatcc
     2761 agatttgaca ccaatattaa ctttttacta gaagaatatg gaatcatggt taataatgat
     2821 gctgtggtta gaaatgtata tcacaaatat ttccatccta aagaagctct agtttccagt
     2881 ggagtcttga acagggaaat tagccgagct gcaggaaagg ctgtgcctgg gatcattgat
     2941 gaggaaagca gtggaaacaa tgcccaggct ctcacctttg tgtatccttt tggtgccaca
     3001 ttgagtgtca tgaaaccagc agtggcggtt ctgtctacag gttctgtctg cttcccactt
     3061 aacagaccca ttttggcttt ctatcactca aagaaccaag gtgggaagct ggcagtgctt
     3121 ggttcatgtc acatgttcag tgatcaatat ttggacaaag aagaaaacag caaaatcatg
     3181 gatgttgttt tccagtggct cacgacagga gacatccacc taaaccagat tgatgctgag
     3241 gacccagaga tttctgacta catgatgctg ccctacacag ccaccctatc aaagcggaat
     3301 cgagagtgtc tccaggagag tgatgagatc ccaagggact ttaccaccct cttcgacctg
     3361 tccatcttcc agctggatac cacctccttc cacagcgtca tcgaggctca cgagcagcta
     3421 aatgtgaaac atgaaccact ccagctcatc cagcctcagt ttgagacgcc gctgccaacc
     3481 cttcagcctg cggtttttcc tcccagtttc cgggagttac cacctcctcc tctggagcta
     3541 tttgatttag atgaaacgtt ctcctctgag aaggcacggc tggctcagat taccaataag
     3601 tgtactgaag aagacctgga attttatgtc aggaagtgtg gtgatattct tggagtaacc
     3661 agtaaactac caaaggacca acaggatgcc aaacatatcc ttgagcacgt cttcttccaa
     3721 gtggtggagt tcaagaaatt gaaccaggaa catgacatcg atacaagtga aacagcattc
     3781 cagaacaatt tctgagaatt cgagctctcg cggccgctcg agggtaccgc gggcccggga
     3841 tccaccggat ctagataact gatcacaatt cactcctcag gtgcaggctg cctatcagaa
     3901 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc gatctttttc
     3961 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     4021 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     4081 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     4141 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     4201 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     4261 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     4321 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     4381 ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag
     4441 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4501 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4561 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     4621 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4681 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4741 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4801 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4861 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4921 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     4981 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5041 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5101 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5161 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5221 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5281 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5341 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5401 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5461 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5521 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5581 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5641 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5701 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5761 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5821 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5881 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5941 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6001 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6061 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6121 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6181 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6241 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6301 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6361 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6421 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6481 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6541 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6601 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6661 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6721 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6781 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6841 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6901 ccgcgcacat ttccccgaaa agtgccacct gggtcgagca gtgtggtttt caagaggaag
     6961 caaaaagcct ctccacccag gcctggaatg tttccaccca atgtcgagca gtgtggtttt
     7021 gcaagaggaa gcaaaaagcc tctccaccca ggcctggaat gtttccaccc aatgtcgagc
     7081 aaaccccgcc cagcgtcttg tcattggcga attaattcga acacgcagat gcagtcgggg
     7141 cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag
     7201 cgaccctgca gccaatatgg gatcggccat tgaacaagat ggattgcacg caggttctcc
     7261 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     7321 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     7381 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     7441 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     7501 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     7561 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     7621 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     7681 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     7741 caggctcaag gcgcgcatgc ccgacggcga tgatctcgtc gtgacccatg gcgatgcctg
     7801 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     7861 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     7921 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     7981 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ggggatcggc aataaaaaga
     8041 cagaataaaa cgcacgggtg ttgggtcgtt tgttcggatc c
//
