LOCUS       sequence-182276-        6185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cilia-membrane-targeted G-actin-binding protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6185)
  AUTHORS   Phua SC, Chiba S, Suzuki M, Su E, Roberson EC, Pusapati GV, Setou M,
            Rohatgi R, Reiter JF, Ikegami K, Inoue T
  TITLE     Dynamic Remodeling of Membrane Composition Drives Cell Cycle through
            Primary Cilia Excision.
  JOURNAL   Cell. 2017 Jan 12;168(1-2):264-279.e15. doi: 
            10.1016/j.cell.2016.12.032. Epub 2017 Jan 12.
  PUBMED    28086093
REFERENCE   2  (bases 1 to 6185)
  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: "Cell."; 
            date: "2017-01-12"; volume: "168(1-2):264-279.e15. doi"; pages: " 
            10.1016/j.cell.2016.12.032. Epub 2017 Jan 12"
COMMENT     Sequence Label: 5HT6-YFP-ThymosinBeta4(WT)
FEATURES             Location/Qualifiers
     source          1..6185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        84..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     542..562
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      620..629
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1976..1985
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1982..1984,1985..1987,1988..2698)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2027..2048)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2288..2307)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2635..2656
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2996..3117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3033..3052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3087..3106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3124..3579)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3261..3282)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3473..3492
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3606..3710
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3712..4069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3712..3903
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3735..3755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3920..4055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4104..4898
                     /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             4104..4898
                     /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(4158..4177)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4768..4787
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5086..5105)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5130..5177
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5506..6094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5995..6014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
      601 aaccgtcaga tccgctagcg ccaccatggt tccagagccc ggccctgtca acagtagcac
      661 cccagcctgg ggccccgggc caccgccagc cccagggggc agcggctggg tggccgctgc
      721 gctgtgcgtg gtcatcgtgc tgacggcagc cgccaattcg ctgctgatcg cgctcatctg
      781 cacgcagccc gcgctgcgca acacgtctaa cttcttcctg gtgtcgctct tcacgtcgga
      841 cctgatggtg ggattggtgg tgatgccccc agccatgctg aacgcgctgt atgggcgctg
      901 ggtactagct cgcggcctct gtctgctttg gaccgccttc gacgtgatgt gctgcagcgc
      961 ctccattctc aacctctgcc tcatcagcct ggaccgctac ctgctcatcc tctcgccgct
     1021 gcgctacaag ctgcgcatga cagccccgcg tgccctggcg ctcatcctgg gtgcctggag
     1081 cctcgccgcg ctggcctcct tcctgcccct cttgctgggc tggcatgaac tgggcaaagc
     1141 tcgaacatct gccccgggcc agtgccgcct attggccagc ctgccttatg tcctcgtggc
     1201 gtccggcgtc acctttttcc tgccttcggg tgccatctgc ttcacctact gcaggatctt
     1261 gctggctgcc cgcaagcagg cggtgcaagt ggcctcgctc accacgggca cggctacggc
     1321 tggccaggcc ttggaaacct tgcaggtgcc caggacacca cgcccaggga tggagtctgc
     1381 tgacagtagg cgtctgacca ccaagcatag caggaaggcc ttgaaggcca gcctgaccct
     1441 cggcatcctg ctgagcatgt tctttgtcac ctggctgccc ttctttgtgg ccagcatagc
     1501 tcaggccgta tgtgactgca tctctccagg tctcttcgat gtcctcacat ggctgggata
     1561 ctgtaatagc accatgaacc ctatcatcta tcccctcttc atgcgggact tcaagagggc
     1621 cctgggcagg ttcgtgccgt gtgtccactg tcccccggag caccgggcca gccccgcctc
     1681 cccctccatg tggacctctc acagtggtgc caggccaggc ctcagcctgc agcaggtgct
     1741 gcccctgcct ctgccaccca actcagattc agactcagct tcagggggca cctcgggcct
     1801 gcagctcaca gcccagcttt tgctgcctgg agaggcgacc cgggaccccc cgccacccac
     1861 cagggcccct actgtggtca acttcttcgt cacagactct gtggagcctg agatacggca
     1921 gcatccactt ggttccccca tgaacagtgc tggtggtagc gcaggaggac cggtcgccac
     1981 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2041 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2101 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2161 cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
     2221 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2281 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2341 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2401 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2461 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2521 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2581 ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2641 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc
     2701 cggactcaga tctcgagctc aagcttcgaa ttcgatgtct gacaaacccg atatggctga
     2761 gatcgagaaa ttcgataagt cgaagttgaa gaaaacagaa acgcaagaga aaaatcctct
     2821 gccttcaaaa gaaacaattg aacaagagaa gcaagctggc gaatcgggat ccaccggatc
     2881 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     2941 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3001 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3061 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     3121 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3181 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3241 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3301 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3361 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3421 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3481 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3541 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3601 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3661 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     3721 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3781 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3841 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3901 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3961 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4021 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4081 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4141 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4201 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4261 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4321 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4381 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4441 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4501 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4561 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4621 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4681 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4741 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4801 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4861 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4921 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4981 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5041 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5101 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5161 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5221 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5281 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5341 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5401 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5461 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5521 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5581 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5641 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5701 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5761 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5821 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5881 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5941 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6001 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6061 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6121 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6181 tgtgg
//
