LOCUS       sequence-434341-        6886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally EGFP-tagged BBS2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6886)
  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 6886)
  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-BBS2
FEATURES             Location/Qualifiers
     source          1..6886
                     /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    3674..3795
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3711..3730)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3765..3784
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3802..4257)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3939..3960)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4151..4170
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4284..4388
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4390..4747
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4390..4581
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4413..4433)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4598..4733
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4782..5576
                     /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             4782..5576
                     /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(4836..4855)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5446..5465
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5764..5783)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5808..5855
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6184..6772
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6673..6692
                     /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 ctgctgcctg tgttcaccct gaaactgcgc
     1381 cacaaaatca gcccccgaat ggtggccata gggcgctacg acgggactca cccgtgcctg
     1441 gcggccgcca cccaaacggg caaggttttt attcataatc ctcatacacg gaaccagcat
     1501 gtcagtgcat ccagggtctt ccagagcccc ctggaatctg atgtttctct tctcaacatt
     1561 aaccaggcag tcagctgtct gactgcaggc gtattgaacc ctgagcttgg ctatgatgcc
     1621 cttttagtgg ggacacagac taatcttttg gcttatgatg tctacaataa ttcggatttg
     1681 ttctacagag aggtagcaga tggggcaaat gcaattgtgc tggggacatt gggagacatt
     1741 tcttcccctc ttgcgattat tggtggcaat tgtgctctgc aaggtttcaa tcatgaagga
     1801 agtgatctct tttggacggt tactggagac aatgttaatt ccttggcctt gtgtgacttt
     1861 gatggtgatg gaaagaaaga gcttcttgtt ggatctgagg attttgatat ccgagttttt
     1921 aaggaagatg agattgtggc agaaatgaca gaaacagaga tagtcacctc tctttgtccc
     1981 atgtatggca gtcgatttgg ttatgccctt tccaatggca cagttggagt ttatgacaaa
     2041 acatcccgat actggagaat taaatcgaaa aatcatgcca tgagcattca tgcttttgac
     2101 cttaattctg atggagtgaa tgaactgata actggttggt ccaatgggaa ggttgatgct
     2161 cgaagtgacc gaactgggga ggtcatcttt aaggacaatt tttcttctgc aattgccggt
     2221 gtggtagagg gagattaccg gatggatggc cacatacagt taatctgctg ctcagtggat
     2281 ggggaaatcc ggggctacct gcctggcacg gctgagatga ggggcaacct catggacacc
     2341 agtgcagagc aggacctgat ccgagagctg agtcagaaga agcagaatct gttgctggaa
     2401 ctccgtaact atgaggaaaa tgccaaggct gaattggcca gtccactgaa cgaggctgat
     2461 gggcatcggg gcataatccc agccaatacc aggctccaca ccacgctctc agtcagcctg
     2521 gggaatgaga cccaaactgc tcatacagaa ttacgcattt ccacttctaa tgacaccatc
     2581 atccgagcag tattgatttt tgcagaagga atttttacag gtgaaagcca cgtggtacat
     2641 cccagcattc acaacctctc cagttccatc tgcatcccta ttgtgcctcc caaagatgtc
     2701 cctgtggatc tgcacttgaa ggcattcgtg ggttacagaa gcagcaccca gtttcatgta
     2761 tttgaatcga caagacagct ccctcgattc tccatgtatg cgctgaccag cctggaccct
     2821 gccagtgagc caatcagtta tgttaacttt accattgcag aacgggcaca gagggttgtt
     2881 gtatggctcg gtcagaactt tctgttacca gaagacactc acattcagaa tgctccattt
     2941 caagtgtgtt tcacatcttt acggaatggc ggccacctgc atataaaaat aaaacttagt
     3001 ggagagatca ctataaatac tgatgatatt gatttggctg gtgatatcat ccagtcaatg
     3061 gcatcatttt ttgctattga agaccttcaa gtagaagcgg attttcctgt ctattttgag
     3121 gaattacgaa aggtgctagt taaggtggat gaatatcatt cagtgcatca gaagctcagt
     3181 gctgatatgg ctgatcattc taatttgatc cgaagtttgc tggtcggagc tgaggatgct
     3241 cgtctgatga gggacatgaa aacaatgaag agtcgttata tggaactcta tgaccttaat
     3301 agagacttgc taaatggata taaaattcgc tgtaacaatc acacagagct gttgggaaac
     3361 ctcaaagcag taaatcaagc aattcaaaga gcaggtcgtc tgcgggttgg aaaaccaaag
     3421 aaccaggtga tcactgcttg tcgggatgca attcgaagca ataacatcaa cacactgttc
     3481 aaaatcatgc gagtggggac agcttcttcc taggaattct gcagtcgacg gtaccgcggg
     3541 cccgggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     3601 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3661 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3721 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3781 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3841 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3901 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3961 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     4021 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4081 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4141 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4201 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     4261 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4321 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4381 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4441 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4501 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4561 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4621 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4681 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4741 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4801 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4861 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4921 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     4981 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5041 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5101 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5161 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5221 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5281 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5341 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5401 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5461 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5521 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5581 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5641 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5701 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     5761 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     5821 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     5881 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5941 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     6001 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     6061 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6121 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6181 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6241 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6301 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6361 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6421 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6481 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6541 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6601 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6661 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6721 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6781 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6841 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
