LOCUS       sequence-177790-        5814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cyan color variant of bright luminescent protein, fused to clathrin 
            light chain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5814)
  AUTHORS   Suzuki K, Kimura T, Shinoda H, Bai G, Daniels MJ, Arai Y, Nakano M, 
            Nagai T
  TITLE     Five colour variants of bright luminescent protein for real-time 
            multicolour bioimaging.
  JOURNAL   Nat Commun. 2016 Dec 14;7:13718. doi: 10.1038/ncomms13718.
  PUBMED    27966527
REFERENCE   2  (bases 1 to 5814)
  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: "Nat 
            Commun."; date: "2016-12-14"; volume: "7:13718. doi"; pages: " 
            10.1038/ncomms13718"
COMMENT     Sequence Label: CeNL-Clathrin_pEGFP
FEATURES             Location/Qualifiers
     source          1..5814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(41..62)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(302..321)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     649..670
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1994..2115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2031..2050)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2085..2104
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2122..2577)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2259..2280)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2471..2490
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2604..2708
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2710..3067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2710..2901
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2733..2753)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2918..3053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3102..3896
                     /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             3102..3896
                     /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(3156..3175)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3766..3785
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4084..4103)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4128..4175
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4504..5092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4993..5012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5267..5570
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5571..5774
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5725..5745
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      5804..5813
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5810..5812,5813..5814,1,2..514)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
ORIGIN
        1 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
       61 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
      121 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
      181 gaccaccctg tcctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
      241 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
      301 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
      361 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
      421 ggagtacaac tactttagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat
      481 caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca
      541 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
      601 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
      661 ggagttcgtg accgccgccg ggttgcatac actcgaagat ttcgttgggg actggcgaca
      721 gacagccggc tacaacctgg accaagtcct tgaacaggga ggtgtgtcca gtttgtttca
      781 gaatctcggg gtgtccgtaa ctccgatcca aaggattgtc ctgagcggtg aaaatgggct
      841 gaagatcgac atccatgtca tcatcccgta tgaaggtctg agcggcgacc aaatgggcca
      901 gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat catcacttta aggtgatcct
      961 gcactatggc acactggtaa tcgacggggt tacgccgaac atgatcgact atttcggacg
     1021 gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc actgtaacag ggaccctgtg
     1081 gaacggcaac aaaattatcg acgagcgcct gatcaacccc gacggctccc tgctgttccg
     1141 agtaaccatc aacggagtga ccggctggcg gctgtgcgaa cgcattctgg cgagatctcg
     1201 agctcaagct tcgaattctg cagtcgacat ggctgatgac tttggcttct tctcgtcgtc
     1261 ggagagtggt gccccggagg cggcggagga ggacccggcg gccgccttcc tggcccagca
     1321 ggagagcgag attgcaggca tagagaacga cgagggcttc ggggcacctg ccggcagcca
     1381 tgcggccccc gcacagccgg gccccacgag tggggctggt tctgaggaca tggggaccac
     1441 agtcaatgga gatgtgtttc aggaggccaa cggtcctgct gatggctacg cagccattgc
     1501 ccaggctgac aggctgaccc aggagcctga gagcatccgc aagtggcgag aggagcagag
     1561 gaaacggctg caagagctgg atgctgcatc taaggtcacg gaacaggaat ggcgggagaa
     1621 ggccaagaag gacctggagg agtggaacca gcgccagagt gaacaagtag agaagaacaa
     1681 gatcaacaac cgggcatccg aggaggcttt cgtgaaggaa tccaaggagg agaccccagg
     1741 cacagagtgg gagaaggtgg cccagctatg tgacttcaac cccaagagca gcaagcagtg
     1801 caaagatgtg tcccgcctgc gctcggtgct catgtccctg aagcagacgc cactgtcccg
     1861 ctagggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     1921 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     1981 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2041 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2101 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2161 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2221 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2281 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2341 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2401 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2461 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2521 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     2581 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2641 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2701 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     2761 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2821 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2881 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2941 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3001 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3061 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3121 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3181 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3241 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     3301 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3361 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3421 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3481 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3541 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3601 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     3661 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3721 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3781 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3841 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3901 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3961 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4021 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4081 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4141 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4201 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4261 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4321 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     4381 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4441 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4501 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4561 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4621 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4681 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4741 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4801 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4861 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4921 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4981 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5041 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5101 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5161 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     5221 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     5281 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     5341 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     5401 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     5461 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     5521 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     5581 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     5641 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     5701 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     5761 atataagcag agctggttta gtgaaccgtc agatccgcta gccgccacca tggt
//
