LOCUS       sequence-339937-        4734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP11-clathrin light chain in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4734)
  AUTHORS   Kamiyama D, Sekine S, Barsi-Rhyne B, Hu J, Chen B, Gilbert LA, 
            Ishikawa H, Leonetti MD, Marshall WF, Weissman JS, Huang B
  TITLE     Versatile protein tagging in cells with split fluorescent protein.
  JOURNAL   Nat Commun. 2016 Mar 18;7:11046. doi: 10.1038/ncomms11046.
  PUBMED    26988139
REFERENCE   2  (bases 1 to 4734)
  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: "2016-03-18"; volume: "7:11046. doi"; pages: " 
            10.1038/ncomms11046"
COMMENT     Sequence Label: pEGFP-GFP11-Clathrin light chain
FEATURES             Location/Qualifiers
     source          1..4734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             598..645
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     polyA_signal    1462..1583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1499..1518)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1553..1572
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1590..2045)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1727..1748)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1939..1958
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2072..2176
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2178..2535
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2178..2369
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2201..2221)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2386..2521
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2570..3364
                     /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             2570..3364
                     /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(2624..2643)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3234..3253
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3552..3571)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3596..3643
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3972..4560
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4461..4480
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatgtcggg aggttccggt ggaagcggag gtagcggcgg atccatgcgt
      601 gaccacatgg tccttcatga gtatgtaaat gctgctggga ttacaggcgg atccggactc
      661 agatctcgag ctcaagcttc gaattctgca gtcgacatgg ctgatgactt tggcttcttc
      721 tcgtcgtcgg agagtggtgc cccggaggcg gcggaggagg acccggcggc cgccttcctg
      781 gcccagcagg agagcgagat tgcaggcata gagaacgacg agggcttcgg ggcacctgcc
      841 ggcagccatg cggcccccgc acagccgggc cccacgagtg gggctggttc tgaggacatg
      901 gggaccacag tcaatggaga tgtgtttcag gaggccaacg gtcctgctga tggctacgca
      961 gccattgccc aggctgacag gctgacccag gagcctgaga gcatccgcaa gtggcgagag
     1021 gagcagagga aacggctgca agagctggat gctgcatcta aggtcacgga acaggaatgg
     1081 cgggagaagg ccaagaagga cctggaggag tggaaccagc gccagagtga acaagtagag
     1141 aagaacaaga tcaacaaccg ggcatccgag gaggctttcg tgaaggaatc caaggaggag
     1201 accccaggca cagagtggga gaaggtggcc cagctatgtg acttcaaccc caagagcagc
     1261 aagcagtgca aagatgtgtc ccgcctgcgc tcggtgctca tgtccctgaa gcagacgcca
     1321 ctgtcccgct agggatccac cggatctaga taactgatca taatcagcca taccacattt
     1381 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     1441 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     1501 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     1561 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     1621 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     1681 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     1741 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1801 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     1861 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     1921 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     1981 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2041 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2101 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2161 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     2221 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2281 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2341 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2401 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2461 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2521 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     2581 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     2641 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     2701 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     2761 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     2821 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     2881 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     2941 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3001 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3061 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     3121 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3181 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3241 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3301 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3361 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     3421 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3481 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     3541 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3601 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     3661 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3721 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     3781 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     3841 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3901 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3961 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4021 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4081 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4141 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4201 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4261 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4321 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4381 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4441 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4501 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4561 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4621 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
     4681 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccg
//
