LOCUS       sequence-434330-        8066 bp    DNA     circular SYN 10-DEC-2025
DEFINITION  Expresses N-terminally EGFP-tagged IFT57 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8066)
  AUTHORS   Funabashi T, Katoh Y, Okazaki M, Sugawa M, Nakayama K
  TITLE     Interaction of heterotrimeric kinesin-II with IFT-B-connecting 
            tetramer is crucial for ciliogenesis.
  JOURNAL   J Cell Biol. 2018 Aug 6;217(8):2867-2876. doi: 
            10.1083/jcb.201801039. Epub 2018 Jun 14.
  PUBMED    29903877
REFERENCE   2  (bases 1 to 8066)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1083/jcb.201801039"; journalName: "J Cell Biol"; date: 
            "2018-08-6- 6"; volume: "217"; issue: "8"; pages: "2867-2876"
COMMENT     Sequence Label: pEGFP-C1-IFT57
FEATURES             Location/Qualifiers
     source          1..8066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        760..1201
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    760..877
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     1172..1195
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1521..1532
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     RBS             1695..1706
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1713..2426
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2438..2509
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2525..2552
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(4006..4594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4086..4105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4808..4827)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4819..5634
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4819..5634
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4892..4911)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     gene            complement(6271..7845)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     terminator      complement(6271..6468)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(6473..7501)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWARGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
ORIGIN
        1 ggagctcaat tctcttagga ttcgattcac attcatcttt ttttagctac taataggata
       61 aattatagga atttataaca aattgaaata tggcaggcag caaaattaaa attgtcttag
      121 ttttttagtg tatagaagtg aatagctata taaagtatgt gtaaagttgg taacggaacg
      181 aaaaatagaa aaggatatta catgggaaaa catgttgttt acggagaaat gaaaagtata
      241 ttgtattttg tacgagctaa aagtacagtg ggaacaaagt cgattttgtt acatctacac
      301 tgttgttatc agatctcgag ctattactag ccgcggtacc aagcttatga gaccatgcat
      361 gcttagtctt aagctgcagg gtctcttgct aaaactgaag caataacaga aaatattgaa
      421 aaacagcgaa gtaactgcat ggatccacta gtatagaagt atagtaattt atgctgcaaa
      481 ggtcctaatg tataaggaaa gaagagaaaa gaagaaaaca agagttttat atacatacag
      541 agcacatgct ctagaccctg ggtgtatata cagttatgct gtgagtatat acagcaccta
      601 gggagctcga aaagaaatat agtcttcaag gttatattat acaggtttat atattattgc
      661 atgtttaaac gaattctagc cccattatct tagcctaaaa aaaccttctc tttggaactt
      721 tcagtaatac gcttaactgc tcattgctat attgaagtac ggattagaag ccgccgagcg
      781 ggcgacagcc ctccgacgga tgactctcct ccgtgcgtcc tcgtcatcac cggtcgcgtt
      841 cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt ctacaatact
      901 agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa accttcaaat
      961 taacgaatca aattaacaac cataggatga taatgcgatt agttttttag ccttatttct
     1021 ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat aaatggaaaa
     1081 gctgcataac cactttaact aatactttca acattttcag tttgtattac ttcttattca
     1141 aatgtcataa aagtatcaac aaaaaattgt taatatacct ctatacttta acgtcaagga
     1201 gaaaaaacta tacccggatt ctagagagct cactagtatc gatgcagtta cttcgctgtt
     1261 tttcaatatt ttctgttatt gcttcagttt tagcacagga actgacaact atatgcgagc
     1321 aaatcccctc accaacttta gaatcgacgc cgtactcttt gtcaacgact actattttgg
     1381 ccaacgggaa ggcaatgcaa ggagtttttg aatattacaa atcagtaacg tttgtcagta
     1441 attgcggttc tcacccctca acaactagca aaggcagccc cataaacaca cagtatgttt
     1501 ttaaggacaa tagctcgacg attgaaggta gatcgagacg ggatccgaaa gtgaaacgtg
     1561 atttcatgcg tcattttgaa cattttgtaa atcttattta ataatgtgtg cggcaattca
     1621 catttaattt atgaatgttt tcttaacatc gcggcaactc aagaaacggc aggttcggat
     1681 cttagctact agagaaagag gagaaatact agatgcgtaa aggcgaagag ctgttcactg
     1741 gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc
     1801 gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta
     1861 ctggtaaact gccggttcct tggccgactc tggtaacgac gctgacttat ggtgttcagt
     1921 gctttgctcg ttatccggac catatgaagc agcatgactt cttcaagtcc gccatgccgg
     1981 aaggctatgt gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg
     2041 cggaagtgaa atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact
     2101 ttaaagagga cggcaatatc ctgggccata agctggaata caattttaac agccacaatg
     2161 tttacatcac cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca
     2221 acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg
     2281 atggtcctgt tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag
     2341 atccgaacga gaaacgcgat catatggttc tgctggagtt cgtaaccgca gcgggcatca
     2401 cgcatggtat ggatgaactg tacaaatgac caggcatcaa ataaaacgaa aggctcagtc
     2461 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     2521 cactggctca ccttcgggtg ggcctttctg cgtttatatt aattaacgtc tcacaatcca
     2581 tggtctatac atgggtgacc aaaagagcgg gcggatacac gcgtcaccac aagcagaata
     2641 aaaggtaaac ctgaaattgt tttaacataa aatgaaaaat gcttgtttgc aaccctatat
     2701 agaatcataa aacattcgtg actataaaat gaataaacta aactattcta agaaaatgaa
     2761 ataaatgaca aaaaaacgtg ttttttggac tagaaggctt aatcaaaagc tcgttcgtct
     2821 gcaattatcg gccagttcag tgagaaggta ttgcgacgaa ttgccacgtt gtcgcagtgt
     2881 ctgtactgtg ggttaatgtc tgtgatcgcc gtcgtgagtt cgtgtcatcc gcgcatgatt
     2941 cagatggtcc tggagatcgt tgcttgtacc ggctgtttgt ggctctacca tctagatgcg
     3001 aattcagggt ggatgggggt aatagaattg tatctatgta tctgacgacc ctgtattaca
     3061 cgaaggaaat agtagatgaa aagacaagag agcaagaaaa aggaaaaact tcctttctaa
     3121 cagacgcttt acttaacctt atatatatct tatttttttc atcgagcgtg ttcaattgga
     3181 caaggtgcca ccttttcgac acttcggtta ttatgttaca cagttttcat gaggatggcg
     3241 ccttgactaa cttaattagt catttgccaa ccaccacagt tccccaatat cgacagcttc
     3301 acgtgccatt tgccattttg cgatcatgtg acctcaagag aaaaagcaaa aaataatgag
     3361 agctaaggga ttcgaaccct tgcatccgaa gatatcagag attttagagg ttaaatccac
     3421 ctaaatctga cgccttaaac cactcggcca aactcccatt gtatgttgga agaattggat
     3481 ttaaatgtct attatagcat taaatatctt taaatacatt caagaaatgg cgaaattatt
     3541 tcttagtgtt ggaataaaaa tcaactatca tctactaact agtatttacg ttactagtat
     3601 attatcatat acggtgttag aagatgacgc aaatgatgag aaatagtcat cttgttggaa
     3661 taaaaatcaa ctatcatcta ctaactagta tttacgttac tagtatatta tcatatacgg
     3721 tgttagaaga tgacgcaaat gatgagaaat agtcatctaa attagtggaa gctgaaacgc
     3781 aaggattgat aatgtaatag gatcaatgaa tattaacata taaaatgatg ataataatat
     3841 ttatagaatt gtgtagaatt gcagattccc ttttatggat tcctaaatcc tcgaattggc
     3901 ggccgcacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3961 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4021 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4081 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4141 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4201 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4261 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4321 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4381 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4441 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4501 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4561 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4621 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga ctgatccttc
     4681 aactcagcaa aagttcgatt tattcaacaa agccacgttg tgtctcaaaa tctctgatgt
     4741 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     4801 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc caggccgcga
     4861 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     4921 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg
     4981 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     5041 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     5101 tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct
     5161 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     5221 cctgtttgta attgtccttt taacagcgat cgcgtatttc gcctcgctca ggcgcaatca
     5281 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     5341 gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc
     5401 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     5461 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     5521 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     5581 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     5641 ttggttaatt ggttgtaaca cgcggccgct cggatggaaa gtagcatcca accaaatccc
     5701 gcgaagtttt cctgatttac ataacaagca aatactaaaa acgttaacac acatgtgctt
     5761 ctttattgta aagaaacctc catcatcctc tattgtaact tggtcctaca ataaatttat
     5821 ccagtgtgac agctttatag gaggcgtaaa gtagtctcat gaagtagatg agtcttgttc
     5881 accatatttt tcttctcgac tgcgaattaa cactgttctt tgatgttaga acaatttagg
     5941 ttcaaagtac gagtcacgac atgtaacaag ggaaaaaaag gatattcatt tcaataaagt
     6001 tcgttttata aaagtatagt acttgtgaga gaaagtaggt ttatacataa ttttacaact
     6061 cattacgcac actcatggtt tttggccagc gaaaacagtt tcaaaagatt gctggaagtc
     6121 tgcaataatg tcatcaataa attcgatacc aacagagaca cgaattaagt ccttggtaac
     6181 accagatgcc aacttttctt tgtcatttaa ttgtttgtgg gtagtgaagt atggagcaat
     6241 gactaaggtc ttggcatcac caacatactc cagtatagcg accagcattc acatacgatt
     6301 gacgcatgat attactttct gcgcacttaa cttcgcatct gggcagatga tgtcgaggcg
     6361 aaaaaaaata taaatcacgc taacatttga ttaaaataga acaactacaa tataaaaaaa
     6421 ctatacaaat gacaagttct tgaaaacaag aatcttttta ttgtcagtta ctttattcct
     6481 ttgccctcgg acgagtgctg gggcgtcggt ttccactatc ggcgagtact tctacacagc
     6541 catcggtcca gacggccgcg cttctgcggg cgatttgtgt acgcccgaca gtcccggctc
     6601 cggatcggac gattgcgtcg catcgacccc gcgcccaagc tgcatcatcg aaattgccgt
     6661 caaccaagct ctgatagagt tggtcaagac caatgcggag catatacgcc cggagccgcg
     6721 gcgatcctgc aagctccgga tgcctccgct cgaagtagcg cgtctgctgc tccatacaag
     6781 ccaaccacgg cctccagaag aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg
     6841 cctcgctcca gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca ttgttggagc
     6901 cgaaatccgc gtgcacgagg tgccggactt cggggcagtc ctcggcccaa agcatcagct
     6961 catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt tgccagtgat
     7021 acacatgggg atcagcaatc gcgcatatga aatcacgcca tgtagtgtat tgaccgattc
     7081 cttgcggtcc gaatgggccg aacccgctcg tctggctaag atcggccgca gcgatcgcat
     7141 ccatggcctc cgcgaccggt tgcagaacag cgggcagttc ggtttcaggc aggtcttgca
     7201 acgtgacacc ctgtgcacgg cgggagatgc aataggtcag gctctcgcta aattccccaa
     7261 tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa gtgccgataa acataacgat
     7321 ctttgtagaa accatcggcg cagctattta cccgcaggac atatccacgc cctcctacat
     7381 cgaagctgaa agcacgagat tcttcgccct ccgagagctg catcaggtcg gacacgctgt
     7441 cgaacttttc gatcagaaac ttctcgacag acgtcgcggt gagttcaggc tttttaccca
     7501 tttttgttta tgttcggatg tgatgtgaga actgtatcct agcaagattt taaaaggaag
     7561 tatatgaaag aagaacctca gtggcaaatc ctaacctttt atatttctct acaggggcgc
     7621 ggcgtgggga caattcaacg cgtctgtgag gggagcgttt ccctgctcgc aggtctggag
     7681 cgaggagccg taatttttgc ttcgcgccgt gcggccatca aaatgtatgg atgcaaatga
     7741 ttatacatgg ggatgtatgg gctaaatgta cgggcgacag tcacatcatg cccctgagct
     7801 gcgcacgtca agactgtcaa ggagggtatt ctgggcctcc atgtcgctgg ccgggtgacc
     7861 cggcggggac gaggcaagct aaactgtact gcagtgcact agttggactg ttgtgatatg
     7921 ccatatatgc agtggtgttc gccggcgctg gttccggctg tcttgcttag ttgtgagtcg
     7981 ccaggcagtc atcggtatga tctgtacatg attcgtcagt gtgagcacca ctgacgagca
     8041 gatttccgta tcatgctcag cgaatt
//
