LOCUS       sequence-434353-        6382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally EGFP-tagged IFT56 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6382)
  AUTHORS   Nozaki S, Katoh Y, Terada M, Michisaka S, Funabashi T, Takahashi S, 
            Kontani K, Nakayama K
  TITLE     Regulation of ciliary retrograde protein trafficking by Joubert 
            syndrome proteins ARL13B and INPP5E.
  JOURNAL   J Cell Sci. 2016 Dec 7. pii: jcs.197004.
  PUBMED    27927754
REFERENCE   2  (bases 1 to 6382)
  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 Cell Sci.
            2016 Dec 7. pii: jcs.197004."
COMMENT     Sequence Label: pEGFP-C1-IFT56
FEATURES             Location/Qualifiers
     source          1..6382
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3170..3291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3207..3226)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3261..3280
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3298..3753)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3435..3456)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3647..3666
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3780..3884
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3886..4243
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3886..4077
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3909..3929)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4094..4229
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4278..5072
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4278..5072
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4332..4351)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4942..4961
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5260..5279)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5304..5351
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5680..6268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6169..6188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctatgatg ctttcaaggg ccaaacctgc tgtaggcaga
     1381 ggcgtacagc acactgacaa aagaaagaag aaaggtagga agattccaaa actagaggag
     1441 ctactttcaa aaagagattt cactggagct attaccctgt tggagttcaa acgtcatgtt
     1501 ggggaagaag aagaggatac taatttgtgg attggatatt gtgcctttca cctgggtgac
     1561 tacaagagag ctctggagga atacgaaaat gctacaaaag aggaaaattg taattctgaa
     1621 gtctgggtga acctagcttg cacctacttc tttcttggga tgtataaaca agctgaagca
     1681 gctggattta aagcttcaaa aagccgactc caaaaccgcc tcctcttcca cttggctcac
     1741 aagtttaatg atgagaaaaa attgatgagc tttcatcaaa atcttcagga tgtcacagaa
     1801 gatcaactca gtttggcctc aatccactat atgcgatctc actaccaaga agctatagat
     1861 atatataagc gaatactgct agataacagg gaataccttg cccttaatgt ttatgtggcc
     1921 ctctgctact acaagttgga ttactatgat gtgtctcaag aagttttggc tgtttacctt
     1981 cagcaaattc ctgatagtac catcgcactc aatcttaaag cctgtaacca ttttcgcctt
     2041 tacaatggca gagcagctga ggcagaactc aaaagcttga tggacaatgc ttcttcatcc
     2101 tttgaatttg ctaaagaact catcaggcac aatctggttg ttttccgagg aggtgaaggg
     2161 gctttgcagg ttttacctcc cctagttgat gtcattcctg aagctaggct aaacttggtg
     2221 atttactacc ttcgtcaaga tgatgtacaa gaagcttata acttaattaa ggatctggaa
     2281 cctactactc ctcaggagta tattctcaaa ggagtggtca atgcagccct tggccaggaa
     2341 atgggttcaa gggatcatat gaaaattgcc cagcagttct tccagttggt gggaggatca
     2401 gctagtgaat gtgatacaat accagggagg cagtgcatgg cttcctgttt cttcctgctt
     2461 aagcaatttg atgatgtttt gatttacctc aactcattta agagttactt ctataatgat
     2521 gacatcttta actttaatta tgcccaagcc aaagctgcaa caggcaatac cagtgagggc
     2581 gaagaggcgt tcctcttgat ccaaagtgag aagatgaaaa atgattacat ttacctcagc
     2641 tggttagctc ggtgctatat tatgaataag aaaccaagac tagcctggga actttatctt
     2701 aagatggaaa cctccggcga gtccttcagt ctcttacagc tcattgctaa tgactgctac
     2761 aagatgggcc agttttacta ttctgccaaa gcttttgatg tccttgagag gctggatcct
     2821 aaccctgaat attgggaagg caaacggggt gcctgtgtgg gcattttcca gatgatcata
     2881 gctgggagag aacccaaaga gacccttcga gaagtgctcc atttactgag aagcacaggt
     2941 aacacccaag tagaatacat gatccggatc atgaagaaat gggccaaaga aaacagagtg
     3001 tccatctaag aattctgcag tcgacggtac cgcgggcccg ggatccaccg gatctagata
     3061 actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3121 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3181 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3241 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa
     3301 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3361 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     3421 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     3481 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     3541 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3601 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3661 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3721 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     3781 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3841 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     3901 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3961 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4021 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4081 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4141 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4201 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     4261 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4321 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4381 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4441 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4501 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4561 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4621 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4681 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4741 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4801 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4861 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4921 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4981 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5041 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5101 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5161 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5221 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     5281 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     5341 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     5401 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     5461 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     5521 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5581 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5641 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5701 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5761 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5821 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5881 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5941 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6001 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6061 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6121 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6181 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6241 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6301 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6361 taaccgtatt accgccatgc at
//
