LOCUS       sequence-434142-        6499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally EGFP-tagged BBS1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6499)
  TITLE     Nakayama Lab Plasmids 2024
REFERENCE   2  (bases 1 to 6499)
  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"
COMMENT     Sequence Label: pEGFP-C1-BBS1
FEATURES             Location/Qualifiers
     source          1..6499
                     /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    3287..3408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3324..3343)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3378..3397
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3415..3870)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3552..3573)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3764..3783
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3897..4001
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4003..4360
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4003..4194
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4026..4046)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4211..4346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4395..5189
                     /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             4395..5189
                     /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(4449..4468)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5059..5078
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5377..5396)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5421..5468
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5797..6385
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6286..6305
                     /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 atccatggcc gctgcgtcct catcggattc cgacgcctgc
     1381 ggagctgaga gcaatgaggc caattcgaag tggttggatg cgcactacga cccaatggcc
     1441 aatatccaca ccttttctgc ctgcctagcg ctggcagatt tacatgggga tggggaatac
     1501 aagctggtgg taggggacct tggccctggt gggcagcagc cccgcctgaa ggtgctcaaa
     1561 ggaccactgg tgatgaccga aagcccgcta cctgctctgc cagctgctgc tgccaccttc
     1621 ctcatggagc aacatgagcc ccggacccca gctctggcac ttgcttcagg cccttgtgtc
     1681 tatgtgtata agaatctcag accctacttc aagttcagcc tgccccaatt gcctccaaat
     1741 cctctggaac aagacctttg gaaccaggcc aaagaggacc gaatcgaccc cttaaccctg
     1801 aaggagatgc tggagagcat ccgggagacg gcagaggagc ctttgtccat ccagtcactc
     1861 aggtttctgc agctggagct aagtgaaatg gaggcatttg taaaccaaca caagtccaac
     1921 tccatcaagc ggcagacagt catcaccacc atgaccacct tgaagaagaa cctggctgac
     1981 gaggatgctg tgtcttgcct ggtgctgggc accgagaaca aggagctcct ggtgcttgac
     2041 cccgaggcct tcaccatttt agccaagatg agccttccca gcgtccccgt cttcctagag
     2101 gtttctggcc agtttgatgt tgagttccgg cttgccgcgg cctgccgcaa tggaaacatc
     2161 tatattctga gaagagactc caagcacccc aagtactgca tcgagctgag cgcccagcct
     2221 gtgggactta tccgggtaca caaggtccta gtggtgggca gcacccaaga cagcctgcat
     2281 ggcttcaccc acaaggggaa gaagctgtgg acagtgcaga tgcccgcagc catcctgacc
     2341 atgaacctcc tggagcagca ttcccggggc ctgcaggccg tcatggctgg gctggccaat
     2401 ggagaggtcc gcatttatcg tgacaaggcc ctgctcaatg tcatccacac cccggatgca
     2461 gtgaccagcc tttgctttgg ccggtacggg cgggaggaca acaccctcat catgaccact
     2521 cgaggtggtg gcctgatcat caagatcctg aagcgtacag cagtgtttgt agagggagga
     2581 agtgaggtgg gtcccccacc agcccaggcc atgaaactca atgtgccccg aaagacccgg
     2641 ctttacgtgg atcagacact gcgagagcgg gaggctggca ccgccatgca ccgggccttc
     2701 cagacagacc tatacctgct gcgcctacgt gctgcccgcg cctacctgca ggccctcgag
     2761 tccagcctga gccccctgtc cacgacagcc cgagagccac tcaagctgca cgccgtggtt
     2821 cagggccttg gccccacctt taagctcaca cttcacctgc agaacacctc aacaacccgt
     2881 cctgtcctgg ggctgctggt ctgcttcctg tacaacgagg cgctctattc cctgccccgg
     2941 gccttcttca aggtaccctt gctggtgcca gggctcaact accccctgga gacctttgtg
     3001 gagagtctca gtaacaaggg catctcagac atcatcaagg tgctggtgct tcgagaaggc
     3061 caaagtgcac ccctgctgag tgcccacgtc aacatgcctg ggagcgaggg gctggcggcc
     3121 gcctgagaat tctgcagtcg acggtaccgc gggcccggga tccaccggat ctagataact
     3181 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3241 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3301 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3361 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     3421 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3481 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3541 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3601 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3661 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3721 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3781 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3841 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3901 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3961 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     4021 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4081 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4141 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4201 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4261 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4321 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4381 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4441 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4501 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4561 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4621 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4681 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4741 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4801 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4861 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4921 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4981 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5041 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5101 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5161 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5221 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5281 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5341 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     5401 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     5461 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     5521 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     5581 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     5641 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5701 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5761 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5821 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5881 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5941 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6001 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6061 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6121 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6181 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6241 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6301 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6361 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6421 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6481 ccgtattacc gccatgcat
//
