LOCUS       sequence-434351-        7384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally EGFP-tagged BBS9 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7384)
  AUTHORS   Katoh Y, Nozaki S, Hartanto D, Miyano R, Nakayama K
  TITLE     Multisubunit complex architectures revealed by visible 
            immunoprecipitation (VIP) assay using fluorescent fusion proteins.
  JOURNAL   J Cell Sci. 2015 May 11. pii: jcs.168740.
  PUBMED    25964651
REFERENCE   2  (bases 1 to 7384)
  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.
            2015 May 11. pii: jcs.168740."
COMMENT     Sequence Label: pEGFP-C1-BBS9
FEATURES             Location/Qualifiers
     source          1..7384
                     /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    4172..4293
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4209..4228)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4263..4282
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4300..4755)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4437..4458)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4649..4668
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4782..4886
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4888..5245
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4888..5079
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4911..4931)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5096..5231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5280..6074
                     /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             5280..6074
                     /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(5334..5353)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5944..5963
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6262..6281)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6306..6353
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6682..7270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7171..7190
                     /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 atccaccatg tctttattta aagcccgtga ttggtggtct
     1381 actattctgg gagataaaga agaatttgat caaggctgtt tgtgtctggc taatgttgac
     1441 aatagtggaa atggacaaga taaaataatt gtgggtagct ttatgggata cctaaggatc
     1501 tttagccccc atcctgcaaa aacaggagat ggagctcaag ccgaagattt gcttctagaa
     1561 gtggatctac gagatccagt acttcaagtg gaagtaggaa agtttgtttc aggtaccgaa
     1621 atgctacatt tggctgtgtt acattctaga aaactttgtg tctactctgt ctcaggaacc
     1681 ttgggtaatg tggaacatgg gaaccaatgt cagatgaaat tgatgtatga acataatctt
     1741 cagagaacag cctgcaatat gacctatgga tcatttggtg gtgtaaaagg tcgagattta
     1801 atttgcatcc agtctatgga tgggatgctg atggtatttg agcaggagag ctatgctttt
     1861 ggaagatttc tccctggctt tcttctgcct ggtcctcttg cctacagttc ccgtacagat
     1921 tccttcctta ctgtctcttc ctgccaacaa gtggaaagtt ataagtacca ggtacttgct
     1981 tttgcaacag atgcagataa aaggcaggag actgaacagc aaaaacttgg ttctggaaaa
     2041 agactagttg tggattggac tctaaatatt ggagagcaag cccttgacat atgtattgtc
     2101 tctttcaatc agtcggcatc ctctgttttt gttcttggtg agagaaactt tttttgcctt
     2161 aaggataatg gacaaattcg attcatgaag aagcttgatt ggagcccaag ttgttttctg
     2221 ccatattgct cagtttctga aggaacaata aatactttga ttggaaatca taataacatg
     2281 ctgcatattt atcaagatgt gacactgaag tgggccaccc aacttcccca cattcctgta
     2341 gcagtaagag tgggctgttt gcatgattta aagggagtga tagtcactct gagtgatgat
     2401 ggtcacttgc agtgttcata cctggggaca gatccttctc tgttccaagc tccaaacgtt
     2461 caatctcgag aactaaacta tgatgaactt gatgtagaaa tgaaagaact tcagaaaatc
     2521 atcaaagatg ttaacaaatc acaaggtgtt tggcccatga ctgagagaga agatgacttg
     2581 aacgtttctg tcgtggtttc tcctaacttt gattcagttt ctcaagcgac cgatgttgag
     2641 gtgggaactg accttgtccc ttctgtcacg gtgaaggtca cactgcagaa cagagtgata
     2701 ttgcaaaaaa ccaaattatc agtctacgtg caaccaccat tagaattgac ttgtgatcag
     2761 ttcacctttg aatttatgac accagatttg actagaacag taagcttttc tgtttatctg
     2821 aaaagaagtt atacaccatc agaattggaa ggaaatgctg ttgtttctta ttccagacca
     2881 acagatcgaa atcctgatgg cattccgcga gttatccaat gtaaatttag acttccccta
     2941 aagttaattt gcctaccagg tcagccttca aaaactgcaa gccacaaaat tactattgat
     3001 accaacaaat ctccagtcag tcttcttagt ctcttcccag gttttgccag tcagtcagat
     3061 gatgatcagg tgaatgtaat gggttttcac ttcttaggag gtgctcgaat tactgttctt
     3121 gcttccaaaa cttctcaacg atatcgcatt cagagtgaac aatttgaaga tctttggctc
     3181 ataaccaatg agcttattct tcgccttcaa gaatattttg aaaaacaggg agtcaaagat
     3241 tttgcatgtt ctttttcggg atctataccc cttcaagaat attttgagtt gattgatcat
     3301 cattttgagc tacggataaa tggtgaaaaa ttagaagaac tcttatctga gagagctgta
     3361 caatttcggg ccattcaacg ccggctacta gcaagattca aagataaaac tcctgcccct
     3421 cttcaacacc tggacacctt gttagatgga acctacaagc aggtaattgc tctagcagat
     3481 gcagtggagg aaaaccaagg caatctgttc cagtcattca ccaggctgaa gagtgccacc
     3541 catttggtga ttctgctgat cgcgctgtgg cagaagctta gtgctgacca ggttgctatt
     3601 ctggaagcgg catttctgcc gctacaagaa gacactcaag aattgggctg ggaagaaacg
     3661 gtggatgccg ccatttccca cctgttgaag acttgcctgt cgaagagttc taaggagcag
     3721 gctttgaacc tcaacagcca gctgaacata cccaaagaca caagccaact gaagaaacat
     3781 atcaccttgc tctgcgatag attatccaaa ggtggccgtc tctgcctaag taccgatgca
     3841 gcagccccac agaccatggt catgccaggt ggttgtacta caatcccaga gtcagaccta
     3901 gaagaaagat cagtagaaca agactctaca gaactgttta ccaaccacag acatctcact
     3961 gcagagacac ccaggcctga agtttcaccc ctccaaggag tctcggaata agaattctgc
     4021 agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca
     4081 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     4141 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4201 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4261 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt
     4321 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4381 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4441 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4501 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     4561 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4621 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     4681 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4741 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4801 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4861 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     4921 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4981 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5041 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5101 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5161 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5221 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     5281 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5341 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5401 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5461 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5521 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5581 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5641 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5701 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5761 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     5821 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5881 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5941 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6001 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6061 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     6121 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6181 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6241 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     6301 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     6361 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     6421 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     6481 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     6541 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6601 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6661 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6721 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6781 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6841 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6901 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6961 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7021 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7081 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7141 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7201 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7261 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7321 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     7381 gcat
//
