LOCUS       sequence-434345-        6280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally EGFP-tagged BBS4 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6280)
  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 6280)
  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-BBS4
FEATURES             Location/Qualifiers
     source          1..6280
                     /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    3068..3189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3105..3124)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3159..3178
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3196..3651)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3333..3354)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3545..3564
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3678..3782
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3784..4141
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3784..3975
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3807..3827)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3992..4127
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4176..4970
                     /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             4176..4970
                     /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(4230..4249)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4840..4859
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5158..5177)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5202..5249
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5578..6166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6067..6086
                     /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 gctgaggaga gagtcgcgac gagaactcaa
     1381 tttcctgtat ctactgagtc tcaaaaaccc cggcagaaaa aagctccaga gtttcctatt
     1441 ttggagaagc agaactggtt gattcatctt cattatatcc ggaaagatta tgaagcctgc
     1501 aaggctgtta tcaaagaaca gcttcaagag actcagggat tgtgtgaata tgctatctat
     1561 gtccaagcat tgatatttcg cctagaagga aatatccaag aatccctaga actcttccag
     1621 acatgtgcag ttcttagtcc tcagagtgct gataacctca agcaggtggc cagatcttta
     1681 tttcttttgg gaaaacataa agctgccatt gaagtatata atgaagcagc taaactcaac
     1741 cagaaagatt gggagatcag ccataaccta ggagtttgct acatatacct gaagcagttc
     1801 aacaaggcac aagaccagtt gcacaatgcc ctgaatctta ataggcacga tctgacttat
     1861 ataatgctgg ggaagatcca cttgctggag ggagacttgg acaaggccat tgaagtctac
     1921 aagaaagcag tggagttctc accagaaaat acagagcttc ttacaacttt aggattactc
     1981 tacttacagc tcggcattta ccagaaggca tttgaacatc ttggcaatgc actgacttat
     2041 gaccctacca actacaaggc catcttggca gcaggcagca tgatgcagac ccacggggac
     2101 tttgatgttg ccctcaccaa atacagagtt gtggcttgtg ctgttccaga aagtcctcca
     2161 ctctggaata acattggaat gtgtttcttt ggcaagaaga aatatgtggc ggccatcagc
     2221 tgcctgaaac gagccaacta cttggcaccc ttcgattgga agattctgta taatttgggc
     2281 cttgtccatt tgaccatgca gcagtatgca tcagcttttc attttctcag tgcggccatc
     2341 aacttccagc caaagatggg ggagctctac atgctcttgg cagtggctct gaccaatctg
     2401 gaagatacag aaaatgccaa gagagcctac gcagaagcag tccacctgga taagtgtaac
     2461 cctttagtaa acctgaacta tgctgtgctg ctgtacaacc agggcgagaa gaagaacgcc
     2521 ctggcccaat atcaggagat ggagaagaaa gtcagcctac tcaaggacaa tagctctctg
     2581 gaatttgact ctgagatggt ggagatggct cagaagttgg gagctgctct ccaggttggg
     2641 gaggcactgg tctggaccaa accagttaaa gatcccaaat caaagcacca gaccacttca
     2701 accagcaaac ctgccagttt ccagcagcct ctgggctcta atcaagctct aggacaggca
     2761 atgtcttcag cagctgcata caggacgctc ccctcaggtg ctggaggaac atcccagttc
     2821 acaaagcccc catctcttcc tctggagcca gagcctgcgg tggaatcaag tccaactgaa
     2881 acatcagaac aaataagaga gaaataagaa ttctgcagtc gacggtaccg cgggcccggg
     2941 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     3001 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     3061 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3121 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3181 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3241 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3301 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3361 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3421 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3481 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3541 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3601 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3661 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3721 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3781 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3841 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3901 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3961 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4021 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4081 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4141 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4201 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4261 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4321 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4381 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4441 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4501 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4561 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4621 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4681 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4741 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4801 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4861 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4921 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4981 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5041 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5101 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     5161 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     5221 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5281 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5341 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5401 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5461 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5521 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5581 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5641 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5701 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5761 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5821 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5881 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5941 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6001 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6061 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6121 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6181 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6241 atcccctgat tctgtggata accgtattac cgccatgcat
//
