LOCUS       sequence-182285-        6185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  non-actin-binding control for 5HT6-YFP-ThymosinBeta4(WT).
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(KK18,19EE)
FEATURES             Location/Qualifiers
     source          1..6185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        76..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        380..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      612..621
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1968..1977
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1974..1976,1977..1979,1980..2690)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2019..2040)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2280..2299)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2627..2648
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2988..3109
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3025..3044)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3079..3098
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3116..3571)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3253..3274)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3465..3484
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3598..3702
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3704..4061
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3704..3895
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3727..3747)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3912..4047
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4096..4890
                     /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             4096..4890
                     /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(4150..4169)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4760..4779
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5078..5097)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5122..5169
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5498..6086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5987..6006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
       61 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      601 gatccgctag cgccaccatg gttccagagc ccggccctgt caacagtagc accccagcct
      661 ggggccccgg gccaccgcca gccccagggg gcagcggctg ggtggccgct gcgctgtgcg
      721 tggtcatcgt gctgacggca gccgccaatt cgctgctgat cgcgctcatc tgcacgcagc
      781 ccgcgctgcg caacacgtct aacttcttcc tggtgtcgct cttcacgtcg gacctgatgg
      841 tgggattggt ggtgatgccc ccagccatgc tgaacgcgct gtatgggcgc tgggtactag
      901 ctcgcggcct ctgtctgctt tggaccgcct tcgacgtgat gtgctgcagc gcctccattc
      961 tcaacctctg cctcatcagc ctggaccgct acctgctcat cctctcgccg ctgcgctaca
     1021 agctgcgcat gacagccccg cgtgccctgg cgctcatcct gggtgcctgg agcctcgccg
     1081 cgctggcctc cttcctgccc ctcttgctgg gctggcatga actgggcaaa gctcgaacat
     1141 ctgccccggg ccagtgccgc ctattggcca gcctgcctta tgtcctcgtg gcgtccggcg
     1201 tcaccttttt cctgccttcg ggtgccatct gcttcaccta ctgcaggatc ttgctggctg
     1261 cccgcaagca ggcggtgcaa gtggcctcgc tcaccacggg cacggctacg gctggccagg
     1321 ccttggaaac cttgcaggtg cccaggacac cacgcccagg gatggagtct gctgacagta
     1381 ggcgtctgac caccaagcat agcaggaagg ccttgaaggc cagcctgacc ctcggcatcc
     1441 tgctgagcat gttctttgtc acctggctgc ccttctttgt ggccagcata gctcaggccg
     1501 tatgtgactg catctctcca ggtctcttcg atgtcctcac atggctggga tactgtaata
     1561 gcaccatgaa ccctatcatc tatcccctct tcatgcggga cttcaagagg gccctgggca
     1621 ggttcgtgcc gtgtgtccac tgtcccccgg agcaccgggc cagccccgcc tccccctcca
     1681 tgtggacctc tcacagtggt gccaggccag gcctcagcct gcagcaggtg ctgcccctgc
     1741 ctctgccacc caactcagat tcagactcag cttcaggggg cacctcgggc ctgcagctca
     1801 cagcccagct tttgctgcct ggagaggcga cccgggaccc cccgccaccc accagggccc
     1861 ctactgtggt caacttcttc gtcacagact ctgtggagcc tgagatacgg cagcatccac
     1921 ttggttcccc catgaacagt gctggtggta gcgcaggagg accggtcgcc accatggtga
     1981 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     2041 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     2101 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     2161 ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     2221 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     2281 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     2341 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     2401 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     2461 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     2521 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     2581 gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     2641 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca
     2701 gatctcgagc tcaagcttcg aattcgatgt ctgacaaacc cgatatggct gagatcgaga
     2761 aattcgataa gtcgaagttg gaggaaacag aaacgcaaga gaaaaatcct ctgccttcaa
     2821 aagaaacaat tgaacaagag aagcaagctg gcgaatcggg atccaccgga tctagataac
     2881 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2941 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3001 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3061 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     3121 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3181 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3241 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3301 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3361 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3421 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3481 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3541 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3601 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3661 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3721 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3781 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3841 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3901 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3961 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     4021 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     4081 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4141 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4201 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4261 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4321 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4381 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4441 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4501 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4561 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4621 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4681 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4741 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4801 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4861 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4921 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4981 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5041 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     5101 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     5161 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     5221 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     5281 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     5341 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5401 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5461 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5521 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5581 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5641 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5701 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5761 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5821 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5881 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5941 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6001 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6061 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6121 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6181 accgt
//
