LOCUS       sequence-434318-        7814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally EGFP-tagged IFT46 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7814)
  AUTHORS   Katoh Y, Terada M, Nishijima Y, Takei R, Nozaki S, Hamada H, 
            Nakayama K
  TITLE     Overall architecture of the intraflagellar transport (IFT)-B complex
            containing Cluap1/IFT38 as an essential component of the IFT-B 
            peripheral subcomplex.
  JOURNAL   J Biol Chem. 2016 Mar 15. pii: jbc.M116.713883.
  PUBMED    26980730
REFERENCE   2  (bases 1 to 7814)
  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: "J Biol 
            Chem. 2016 Mar 15. pii: jbc.M116.713883."
COMMENT     Sequence Label: pCAG-EGFP-C-IFT46
FEATURES             Location/Qualifiers
     source          1..7814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        366..643
                     /label=chicken beta-actin promoter
     intron          644..1660
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1584..1606
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1668..1687
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1725..1734
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1731..1733,1734..1736,1737..2447)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1731..1733,1734..1736,1737..2447)
                     /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(1776..1797)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2037..2056)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2384..2405
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3667..3686)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3732..3787
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3786..3805)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4148..4164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4148..4164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4161..4183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4172..4188
                     /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(4196..4202,4203..4220,4221..4226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4241..4262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4321..4516
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4367..4502
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4522..4656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4559..4578)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4613..4632
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4724..4741)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4895..5483)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4975..4994)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5654..6445,6446..6514))
                     /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     6277..6296
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6515..6619)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6931..7734
                     /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(6994..7013)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7604..7623
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7741..7789
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        join(7799..7814,1..364)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      421 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
      481 gggggggcgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag
      541 aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
      601 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg
      661 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
      721 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
      781 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg
      841 ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg
      901 gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg
      961 gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg
     1021 cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
     1081 agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc
     1141 tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt
     1201 gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg
     1261 ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc
     1321 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
     1381 ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg
     1441 ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc
     1501 gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct
     1561 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
     1621 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1681 tattgtgctg tctcatcatt ttggcaaaga atttgctagt tttcgccacc atggtgagca
     1741 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1801 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1861 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1921 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     1981 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2041 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2101 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2161 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     2221 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     2281 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     2341 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2401 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat
     2461 ctatggctga taacagcagt gatgagtgtg aagaggaaaa taacaaggtc ctaagggagg
     2521 gcatgccaca ggctccaggc cacagaggca aagacatgga ccctgttcca cctgcccctg
     2581 caagtctcaa gtgccaccag accccttctc atgtacttga gagggtggga tggtacagag
     2641 agagccagaa gcacagaaag gagaagaaga agacctcaca gttgacacct caacggggct
     2701 ttagtgaaaa tgaggatgac gatgatgatg atgatgattc atctgaaact gattctgatt
     2761 ctgatgatga tgatgaagag catggagccc ctctggaagg ggcctatgac cctgcagact
     2821 atgagcattt gccagtttct gctgaaatta aggaactctt ccagtacatc agtaggtaca
     2881 cacctcagtt gattgacctg gaccacaaac tgaagccttt cattcctgat tttatcccag
     2941 ctgtcgggga tattgatgca ttcttaaagg tcccacgtcc tgatggaaag cctgacaacc
     3001 ttggcctatt ggtattggat gaaccttcta caaagcagtc agaccctacg gtgctctcac
     3061 tctggttaac agagaattct aagcagcaca acatcacaca acatatgaaa gtaaaaagcc
     3121 tagaagatgc agaaaagaat cccaaagcca ttgacacgtg gattgagagc atctctgaat
     3181 tacaccgttc taagccccct gcgactgtgc actacaccag gcccatgccc gacattgaca
     3241 cgctgatgca ggaatggtcc ccggagtttg aagagctttt gggcaaggta agcctgccca
     3301 cggcagagat tgattgcagc ctggcagagt acattgacat gatctgtgcc attctagaca
     3361 tccctgtcta caagagtcgg atccagtccc tccatctgct cttttccctc tactcagaat
     3421 tcaagaactc acagcatttt aaagctctcg ctgaaggcaa gaaagcattc actccttcat
     3481 ccaattccac ctcccaagct ggagacatgg agacattaac cttcagctga ctcgagggta
     3541 ccgcgggccc gggatccacc ggatctagat aactgatcac aattcactcc tcaggtgcag
     3601 gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga
     3661 gatcgatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     3721 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     3781 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     3841 ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata
     3901 aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc
     3961 cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc
     4021 cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc
     4081 agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt
     4141 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4201 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4261 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc
     4321 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4381 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4441 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4501 cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc
     4561 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4621 tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac
     4681 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4741 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4801 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4861 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4921 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4981 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5041 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5101 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5161 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5221 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5281 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5341 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5401 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5461 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5521 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5581 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5641 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5701 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5761 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5821 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5881 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5941 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6001 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6061 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6121 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6181 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6241 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6301 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6361 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6421 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6481 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6541 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6601 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcg agcagtgtgg
     6661 ttttcaagag gaagcaaaaa gcctctccac ccaggcctgg aatgtttcca cccaatgtcg
     6721 agcagtgtgg ttttgcaaga ggaagcaaaa agcctctcca cccaggcctg gaatgtttcc
     6781 acccaatgtc gagcaaaccc cgcccagcgt cttgtcattg gcgaattaat tcgaacacgc
     6841 agatgcagtc ggggcggcgc ggtcccaggt ccacttcgca tattaaggtg acgcgtgtgg
     6901 cctcgaacac cgagcgaccc tgcagccaat atgggatcgg ccattgaaca agatggattg
     6961 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     7021 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     7081 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     7141 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     7201 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     7261 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     7321 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     7381 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     7441 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc
     7501 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     7561 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     7621 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     7681 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaggggat
     7741 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttcg gatccgtcga
     7801 cattgattat tgac
//
