LOCUS       sequence-380171-        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 rigid alpha helix 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(5)
FEATURES             Location/Qualifiers
     source          1..5351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        76..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        380..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    606..686
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1357..1378)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1618..1637)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1777..2028
                     /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     1965..1986
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2154..2275
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2191..2210)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2245..2264
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2282..2737)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2419..2440)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2631..2650
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2764..2868
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2870..3227
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2870..3061
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2893..2913)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3078..3213
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3262..4056
                     /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             3262..4056
                     /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(3316..3335)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3926..3945
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4244..4263)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4288..4335
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4664..5252
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5153..5172
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
       61 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      601 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgacg
      661 gtaccgcggg cccgggatcc accggtcgcc accatggctg atgactttgg cttcttctcg
      721 tcgtcggaga gtggtgcccc ggaggcggcg gaggaggacc cggcggccgc cttcctggcc
      781 cagcaggaga gcgagattgc aggcatagag aacgacgagg gcttcggggc acctgccggc
      841 agccatgcgg cccccgcaca gccgggcccc acgagtgggg ctggttctga ggacatgggg
      901 accacagtca atggagatgt gtttcaggag gccaacggtc ctgctgatgg ctacgcagcc
      961 attgcccagg ctgacaggct gacccaggag cctgagagca tccgcaagtg gcgagaggag
     1021 cagaggaaac ggctgcaaga gctggatgct gcatctaagg tcacggaaca ggaatggcgg
     1081 gagaaggcca agaaggacct ggaggagtgg aaccagcgcc agagtgaaca agtagagaag
     1141 aacaagatca acaaccgggc atccgaggag gctttcgtga aggaatccaa ggaggagacc
     1201 ccaggcacag agtgggagaa ggtggcccag ctatgtgact tcaaccccaa gagcagcaag
     1261 cagtgcaaag atgtgtcccg cctgcgctcg gtgctcatgt ccctgaagca ggaagctgct
     1321 gcaaaggagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1381 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1441 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1501 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1561 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1621 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1681 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1741 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1801 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1861 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1921 acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1981 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc
     2041 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2101 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2161 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2221 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     2281 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2341 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2401 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2461 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2521 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2581 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2641 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2701 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2761 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2821 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     2881 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2941 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3001 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3061 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3121 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3181 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3241 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3301 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3361 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3421 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3481 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3541 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3601 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3661 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3721 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3781 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3841 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3901 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3961 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4021 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4081 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4141 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4201 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4261 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4321 ttgggtcgtt tgttcataaa cgcggggttc ggtcacaggg ctggcactct gtcgataccc
     4381 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4441 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4501 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4561 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4621 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4681 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4741 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4801 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4861 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4921 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4981 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5041 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5101 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5161 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5221 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5281 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5341 tggataaccg t
//
