LOCUS       sequence-182282-        7961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  kinase-dead control for 5HT6-YFP-PIPK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7961)
  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 7961)
  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-PIPK(D253A)
FEATURES             Location/Qualifiers
     source          1..7961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        131..434
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        435..638
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     589..609
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      667..676
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2023..2032
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2029..2031,2032..2034,2035..2745)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2074..2095)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2335..2354)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2682..2703
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4819..4940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4856..4875)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4910..4929
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4947..5402)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5084..5105)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5296..5315
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5429..5533
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5535..5892
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5535..5726
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5558..5578)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5743..5878
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5927..6721
                     /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             5927..6721
                     /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(5981..6000)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6591..6610
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6909..6928)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6953..7000
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7329..7917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7818..7837
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
       61 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      121 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      181 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      241 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      301 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      361 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      421 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      481 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      541 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      601 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      661 gctagcgcca ccatggttcc agagcccggc cctgtcaaca gtagcacccc agcctggggc
      721 cccgggccac cgccagcccc agggggcagc ggctgggtgg ccgctgcgct gtgcgtggtc
      781 atcgtgctga cggcagccgc caattcgctg ctgatcgcgc tcatctgcac gcagcccgcg
      841 ctgcgcaaca cgtctaactt cttcctggtg tcgctcttca cgtcggacct gatggtggga
      901 ttggtggtga tgcccccagc catgctgaac gcgctgtatg ggcgctgggt actagctcgc
      961 ggcctctgtc tgctttggac cgccttcgac gtgatgtgct gcagcgcctc cattctcaac
     1021 ctctgcctca tcagcctgga ccgctacctg ctcatcctct cgccgctgcg ctacaagctg
     1081 cgcatgacag ccccgcgtgc cctggcgctc atcctgggtg cctggagcct cgccgcgctg
     1141 gcctccttcc tgcccctctt gctgggctgg catgaactgg gcaaagctcg aacatctgcc
     1201 ccgggccagt gccgcctatt ggccagcctg ccttatgtcc tcgtggcgtc cggcgtcacc
     1261 tttttcctgc cttcgggtgc catctgcttc acctactgca ggatcttgct ggctgcccgc
     1321 aagcaggcgg tgcaagtggc ctcgctcacc acgggcacgg ctacggctgg ccaggccttg
     1381 gaaaccttgc aggtgcccag gacaccacgc ccagggatgg agtctgctga cagtaggcgt
     1441 ctgaccacca agcatagcag gaaggccttg aaggccagcc tgaccctcgg catcctgctg
     1501 agcatgttct ttgtcacctg gctgcccttc tttgtggcca gcatagctca ggccgtatgt
     1561 gactgcatct ctccaggtct cttcgatgtc ctcacatggc tgggatactg taatagcacc
     1621 atgaacccta tcatctatcc cctcttcatg cgggacttca agagggccct gggcaggttc
     1681 gtgccgtgtg tccactgtcc cccggagcac cgggccagcc ccgcctcccc ctccatgtgg
     1741 acctctcaca gtggtgccag gccaggcctc agcctgcagc aggtgctgcc cctgcctctg
     1801 ccacccaact cagattcaga ctcagcttca gggggcacct cgggcctgca gctcacagcc
     1861 cagcttttgc tgcctggaga ggcgacccgg gaccccccgc cacccaccag ggcccctact
     1921 gtggtcaact tcttcgtcac agactctgtg gagcctgaga tacggcagca tccacttggt
     1981 tcccccatga acagtgctgg tggtagcgca ggaggaccgg tcgccaccat ggtgagcaag
     2041 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2101 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2161 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2221 ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     2281 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2341 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2401 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2461 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2521 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2581 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
     2641 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     2701 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct
     2761 cgagctcaag cttcgaattc gatggagcta gaggtgccgg acgaggcgga gagcgccgaa
     2821 gcgggggccg tgacggcgga agcggcctgg tctgcggaga gtggggcggc ggcaggtatg
     2881 acccagaaga aggctggcct cgcagaggcc cccttggtga cagggcagcc aggccctggc
     2941 catgggaaga agctgggcca ccgaggcgtg gatgcgtcgg gagagactac gtataagaag
     3001 accacctcgt ccaccctgaa gggcgccatc cagctgggga tcgggtacac ggtgggcaac
     3061 ctgagctcta agccggagcg ggacgtgctc atgcaggact tctacgtgat ggagagcatc
     3121 ttcttcccca gtgaagggag caacttcacc cctgcccacc acttccagga tttccgcttc
     3181 aagacctatg cacctgttgc cttccgctac ttccgggagc tctttggcat ccgtccagat
     3241 gactatttgt actccctgtg caatgagcca ctcatcgagc tctccaaccc cggggccagc
     3301 ggctctgtct tctacgtcac cagcgacgac gagttcatca tcaagactgt catgcacaag
     3361 gaggcagagt tcctgcagaa gctgctgcct ggctactaca tgaatctcaa ccagaaccca
     3421 cggacgctgc tgcccaagtt ctatgggctg tactgcgtgc agtctggtgg caagaacatc
     3481 cgcgtggtgg tcatgaacaa tgtgctgccc cgtgtcgtca aaatgcacct taagttcgcc
     3541 ctcaagggct ccacgtacaa gcgcagggcc gccaagaagg agaaggagaa gagcctgccc
     3601 acctacaagg acctggactt catgcaggac atgcccgagg ggctgctgct ggactccgac
     3661 acctttggcg ccctggtcaa gacgctgcag cgagactgcc tggtgctgga gagcttcaag
     3721 ataatggact acagcctgct gctgggcgtg cacaacatcg atcagcagga gcgagagcgc
     3781 caggccgagg gcgcccagag caaggcggat gagaagcggc ccgtggccca gaaggctctg
     3841 tattccacgg ccatggagtc tatccagggc ggagctgccc gtggggaggc cattgagaca
     3901 gatgacacga tgggtgggat tccagcagtg aatgggcgcg gggagcgact gctcctccac
     3961 atcgggatca ttgatattct gcagtcctac aggttcatca agaagttaga acacacctgg
     4021 aaggccctcg tccatgatgg ggacactgtc tcagtccacc ggcccagctt ctatgcagag
     4081 cgcttcttca agttcatgag cagcacggtg ttcgaggaga gttcctccct gaagtcctct
     4141 ccatccaaga aaggacgtgg tgccctgctg gcggtcaaac ccctggggcc cactgctgcc
     4201 ttctcagcca gccagatccc cagcgagaga gaggatgtgc agtatgacct gcggggggcc
     4261 cgcagctacc ccacgcttga ggatgaaggc cggcctgacc tcctgccctg caccccaccg
     4321 tcctttgagg aagccaccac agcctccatc gccaccaccc tgtcgtccac ctccctctcc
     4381 atcccagagc ggtccccttc agatacatcg gagcagcccc ggtacaggcg gcgcacgcag
     4441 tcttcaggcc aggatggccg gccccaggag gagccccatg cggaagacct gcagaagata
     4501 actgtgcagg tggagccagt gtgcggtgtg ggggttgtcc ccaaggagga gggtgcagga
     4561 gtggaggtcc ccccatgtgg ggcatcggct gcagcctctg tggaaataga cgctgccagc
     4621 caggcctcag agcctgccag ccaggcctca gatgaggagg atgcaccctc tacagacatc
     4681 tatttttaag gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     4741 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     4801 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4861 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4921 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     4981 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     5041 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     5101 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     5161 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5221 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5281 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5341 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5401 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5461 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5521 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     5581 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5641 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5701 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5761 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5821 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5881 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     5941 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     6001 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     6061 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     6121 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     6181 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     6241 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     6301 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     6361 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     6421 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     6481 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     6541 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     6601 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     6661 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     6721 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     6781 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6841 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     6901 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     6961 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     7021 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     7081 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     7141 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     7201 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7261 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7321 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7381 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7441 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7501 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7561 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7621 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7681 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7741 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7801 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7861 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7921 gccagcaacg cggccttttt acggttcctg gccttttgct g
//
