LOCUS       sequence-434347-        6858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally EGFP-tagged BBS7 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6858)
  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 6858)
  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-BBS7
FEATURES             Location/Qualifiers
     source          1..6858
                     /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    3646..3767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3683..3702)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3737..3756
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3774..4229)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3911..3932)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4123..4142
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4256..4360
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4362..4719
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4362..4553
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4385..4405)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4570..4705
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4754..5548
                     /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             4754..5548
                     /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(4808..4827)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5418..5437
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5736..5755)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5780..5827
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6156..6744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6645..6664
                     /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 gatctgattt taaaccgaat ggattatctg
     1381 caggtgggag taacatctca gaagactatg aagctaattc ctgcctcaag acacagagct
     1441 acacaaaagg tggttattgg agatcatgat ggggtagtta tgtgctttgg catgaagaaa
     1501 ggagaagcag cagcagtgtt caagacttta cccgggccga agattgcaag gctggaactg
     1561 ggaggggtta tcaacacacc tcaggagaaa atttttattg ctgcagcatc tgagattaga
     1621 ggcttcacaa aaagaggaaa acagttcctc tcctttgaaa caaacctcac tgaaagcatt
     1681 aaagctatgc acatatctgg ctcagacctc tttctcagtg caagttacat ctataaccat
     1741 tattgtgact gcaaagacca acattattac ctttctgggg ataaaatcaa tgatgtgatc
     1801 tgccttccag tggaaagatt atctcgtatc acacctgtat tggcctgcca ggacagagtg
     1861 ctcagagttt tacagggatc tgatgtgatg tatgcagttg aagttcctgg accccctact
     1921 gtcttagcac tacacaatgg aaatggcggt gactctggag aagacctttt gtttgggaca
     1981 tcagacggaa aacttgcgct tatacagatt actacatcca aaccagtacg caagtgggaa
     2041 attcaaaatg agaaaaagag aggaggtatt ttgtgtattg acagctttga cattgtgggt
     2101 gatggggtta aagatttact tgttgggaga gatgacggaa tggtggaagt gtatagtttt
     2161 gataatgcaa atgaacctgt tctacgattt gatcagatgt tgtctgaaag cgtcacatct
     2221 atccagggtg gttgtgtagg aaaagacagc tatgatgaaa tcgtggtgtc cacatattca
     2281 ggctgggtta caggtctgac aacagagccc attcataagg aaagtggacc aggagaagaa
     2341 ctaaaaatta atcaggagat gcagaataaa atttcttcct tacggaatga gttggaacat
     2401 ttgcagtata aggtattgca ggaaagagag aattatcaac agtcttctca atcaagcaaa
     2461 gcaaaatcag cagtaccttc ctttggtata aatgataaat ttacactaaa taaagatgat
     2521 gccagttaca gccttatctt agaggtacag actgcaatag ataatgtctt aatacagagt
     2581 gatgttccaa tagatttact tgatgtggat aaaaattctg ctgttgttag ctttagcagc
     2641 tgtgattctg agtcaaacga caacttcctt cttgccactt atcggtgcca ggcagatact
     2701 acaaggctgg aactcaagat tcgctcaatt gaaggccagt atggcacact acaagcatat
     2761 gtgactccaa gaattcaacc caaaacctgt caggtccgcc agtaccacat caaacctctt
     2821 tcactccatc aaagaactca ctttattgat catgacagac ccatgaatac actgacccta
     2881 acaggccagt tcagttttgc tgaagttcac tcctgggtgg ttttttgtct gcctgaagtt
     2941 ccagaaaaac ctccagcagg agaatgtgtg acattttact ttcagaacac ctttctagat
     3001 acacaacttg aaagtaccta cagaaaagga gagggagttt ttaaatctga caacatttct
     3061 actatctcca tcctaaaaga tgtgctttct aaagaagcta caaaaaggaa aattaacctc
     3121 aacatatcat acgagataaa tgaagtatca gtcaaacaca ctttaaagct aatccaccca
     3181 aagctggagt accagttgct tttggctaag aaagtgcagt taattgatgc tttaaaagaa
     3241 ttacagattc atgagggaaa tacgaacttt ctgataccag aatatcactg tattctagaa
     3301 gaggcagatc acctacagga agaatacaaa aagcaacctg cacatcttga aagactctat
     3361 ggcatgatca ctgatctttt catagataaa tttaagttta aaggcaccaa tgtaaaaact
     3421 aaagtacccc ttctattgga aattctggac agttatgacc aaaatgcatt gatttcattc
     3481 ttcgatgcag catgactcga cggtaccgcg ggcccgggat ccaccggatc tagataactg
     3541 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3601 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3661 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3721 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     3781 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3841 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3901 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3961 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     4021 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     4081 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4141 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4201 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4261 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4321 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4381 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4441 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4501 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4561 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4621 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4681 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4741 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4801 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4861 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4921 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4981 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5041 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5101 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5161 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5221 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5281 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5341 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5401 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5461 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5521 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5581 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5641 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5701 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5761 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5821 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5881 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5941 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     6001 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6061 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6121 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6181 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6241 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6301 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6361 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6421 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6481 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6541 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6601 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6661 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6721 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6781 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6841 cgtattaccg ccatgcat
//
