LOCUS       sequence-434348-        6238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally EGFP-tagged BBS8 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6238)
  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 6238)
  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-BBS8
FEATURES             Location/Qualifiers
     source          1..6238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(39..58)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     249..270
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    414..535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(425..444)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     479..498
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(2232..2942,2943..2945,2946..2948))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2232..2942,2943..2945,2946..2948))
                     /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(2274..2295)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2623..2642
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2882..2903
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2945..2954
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2993..3196)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3022..3042)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3197..3500
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3675..4263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3755..3774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4592..4639
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4664..4683
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4871..5665)
                     /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             complement(4871..5665)
                     /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(4982..5001)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5592..5611
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5700..6057)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5714..5849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5866..6057
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6014..6034
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(6059..6163)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6190..6238,1..407)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
       61 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      121 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      181 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      241 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      301 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      361 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttacg cgttaagata
      421 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
      481 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
      541 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
      601 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagttatcta gatccggtgg
      661 atcccgggcc cgcggtaccg tcgactgcag aattctcaga gcatagcaaa atgctgcctt
      721 aattgtttaa ttaaatgttg tgtgtccaca tggtctggaa atgctgcttc agacttctgc
      781 gcagcaacat agcttctctg cagatctcca atcttatcag agattgttgc aaaattaaaa
      841 tgcggttcat acatatgggg tgctaatgat gatgcagttt gtaatagtgc ccttgcctgt
      901 tcaacgtggc ccttccgcat ctccagcaca gccaggttgt tgtaggcctc ggcgtggttg
      961 ttgttgttga ccagagccag cctgaagcac tgatgggcca aatttgtatc tcctattccc
     1021 acagctacat gtcccaagtt gtaccagaca tcagctgcct cttcttcatt ttcagccaaa
     1081 gaaagggcac gttcaaatga ggtcagagtc atatcatact gctgggcata gaagcaacac
     1141 agccccagat tgttaaaaag ctggccgtta taaatgccca tctgcagcag ccgcctgtaa
     1201 aaccggagag ctatttctgg ctgatcagaa tagaagtggt tgcttccaat gcatgcgatg
     1261 gcttccacat gagtattgtc ttgtttcaaa acttctttgt aatattctgc tgctgatgac
     1321 atattgttca tttcctcata gattcttgca attccacaga gcagggttac ttctcctgga
     1381 aacttatcta agccttgttt gaaaagattt aaagcagtca caggttgatc caatgagaca
     1441 taaacttttg ccaagtacag aaatgtatct accatttcct gctgcttcag ggctgattta
     1501 aactgttttt ctgcttcacg atacattccc aacctgtagt aacattttcc aatctgtact
     1561 ttccaccacc agtccttgta ctgagaatgt tctgtggaga gggcagccag atccaaagca
     1621 gtcttaacat cattttcatg atgaaagata tactcaaaca aagcctttgc caacttaggt
     1681 ttctgggaat actttgttaa attcagccta gataaattta taaatggtcc atcaggactt
     1741 gtaagcatgg aagccgttcc cagcctgaca aatcttccgg aggagctggt gatagggcgg
     1801 gctgtgtagg cggttctggg tgttctgata gcctgttcca tagtgcctgg ccttccactc
     1861 tgcgtgctgg gcctgaggaa acctgtaatg ggtcttccag cttgtgtgat tggcctaacg
     1921 gcctggctag gccctcctgt ctgattagtt ccagggagtt tcaaagacgt tccagggcgt
     1981 ggaacttgag ctatagcatt ttcatccagc atcatttctg caattccttc ctgatctaca
     2041 tcaatttcat ctatgtatac catttctgtt agcgctcttg cttttaagat ccaagctgcc
     2101 tggtcataag gggacttctc cagcatctgc gtgcatagat cggcgcagag ctggaacttc
     2161 ctgcgcctaa aatagctcca ggccaggagc agcggctcca tctccgagct catggtggat
     2221 ctgagtccgg acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac
     2281 tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactgggtg
     2341 ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg
     2401 tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc
     2461 ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac
     2521 tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg
     2581 cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg
     2641 tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag
     2701 tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg
     2761 gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc
     2821 agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt
     2881 acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg
     2941 ctcaccatgg tggcgaccgg tagcgctagc ggatctgacg gttcactaaa ccagctctgc
     3001 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     3061 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     3121 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     3181 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     3241 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     3301 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     3361 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     3421 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     3481 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     3541 ttgattacta ttaataacta atgcatggcg gtaatacggt tatccacaga atcaggggat
     3601 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3661 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3721 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3781 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3841 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3901 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3961 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4021 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4081 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4141 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4201 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4261 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4321 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4381 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct
     4441 gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg
     4501 gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag
     4561 tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta
     4621 ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt
     4681 tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     4741 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     4801 gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     4861 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     4921 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     4981 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     5041 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     5101 catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt
     5161 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     5221 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     5281 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     5341 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     5401 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     5461 acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga
     5521 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     5581 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     5641 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcgatct
     5701 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
     5761 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     5821 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
     5881 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     5941 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     6001 ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcaggac
     6061 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6121 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6181 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcg
//
