LOCUS       sequence-380174-        5351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Clathrin Light Chain B tagged with mEGFP at amino 
            acid position 206 with a flexible linker of 5 amino acids in 
            mammalian cells. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5351)
  AUTHORS   Obashi K, Sochacki KA, Strub MP, Taraska JW
  TITLE     A conformational switch in clathrin light chain regulates lattice 
            structure and endocytosis at the plasma membrane of mammalian cells.
  JOURNAL   Nat Commun. 2023 Feb 9;14(1):732. doi: 10.1038/s41467-023-36304-7.
  PUBMED    36759616
REFERENCE   2  (bases 1 to 5351)
  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: "2023-02-9"; volume: "14(1):732. doi"; pages: " 
            10.1038/s41467-023-36304-7"
COMMENT     Sequence Label: CLC-EGFP206(flex5)
FEATURES             Location/Qualifiers
     source          1..5351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    598..678
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      680..689
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1349..1370)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1610..1629)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1769..2020
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1957..1978
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2146..2267
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2183..2202)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2237..2256
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2274..2729)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2411..2432)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2623..2642
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2756..2860
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2862..3219
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2862..3053
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2885..2905)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3070..3205
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3254..4048
                     /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             3254..4048
                     /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(3308..3327)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3918..3937
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4236..4255)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4280..4327
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4656..5244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5145..5164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg
      661 ggcccgggat ccaccggtcg ccaccatggc tgatgacttt ggcttcttct cgtcgtcgga
      721 gagtggtgcc ccggaggcgg cggaggagga cccggcggcc gccttcctgg cccagcagga
      781 gagcgagatt gcaggcatag agaacgacga gggcttcggg gcacctgccg gcagccatgc
      841 ggcccccgca cagccgggcc ccacgagtgg ggctggttct gaggacatgg ggaccacagt
      901 caatggagat gtgtttcagg aggccaacgg tcctgctgat ggctacgcag ccattgccca
      961 ggctgacagg ctgacccagg agcctgagag catccgcaag tggcgagagg agcagaggaa
     1021 acggctgcaa gagctggatg ctgcatctaa ggtcacggaa caggaatggc gggagaaggc
     1081 caagaaggac ctggaggagt ggaaccagcg ccagagtgaa caagtagaga agaacaagat
     1141 caacaaccgg gcatccgagg aggctttcgt gaaggaatcc aaggaggaga ccccaggcac
     1201 agagtgggag aaggtggccc agctatgtga cttcaacccc aagagcagca agcagtgcaa
     1261 agatgtgtcc cgcctgcgct cggtgctcat gtccctgaag caggggggtg gcggcagcga
     1321 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1381 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1441 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1501 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1561 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1621 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1681 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1741 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     1801 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1861 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1921 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1981 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag
     2041 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2101 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2161 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2221 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     2281 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2341 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2401 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2461 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2521 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2581 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2641 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2701 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2761 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2821 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     2881 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2941 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3001 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3061 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3121 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3181 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3241 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3301 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3361 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3421 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3481 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3541 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3601 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3661 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3721 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3781 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3841 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3901 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3961 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4021 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4081 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4141 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4201 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4261 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4321 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4381 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4441 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     4501 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4561 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4621 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4681 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4741 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4801 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4861 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4921 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4981 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5041 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5101 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5161 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5221 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5281 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5341 cgtattaccg c
//
