LOCUS       sequence-380163-        5423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Clathrin Light Chain B (neuronal isoform) tagged 
            with ShadowY in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5423)
  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 5423)
  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: nCLC-ShadowY
FEATURES             Location/Qualifiers
     source          1..5423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        69..372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     527..547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    599..679
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      681..690
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1422..1443)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1683..1702)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2030..2051
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2219..2340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2256..2275)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2310..2329
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2347..2802)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2484..2505)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2696..2715
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2829..2933
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2935..3292
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2935..3126
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2958..2978)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3143..3278
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3327..4121
                     /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             3327..4121
                     /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(3381..3400)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3991..4010
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4309..4328)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4353..4400
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4729..5317
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5218..5237
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
       61 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      121 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      181 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      241 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      301 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      361 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      421 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      481 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      541 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      601 tagcgctacc ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc
      661 gggcccggga tccaccggtc gccaccatgg ctgatgactt tggcttcttc tcgtcgtcgg
      721 agagtggtgc cccggaggcg gcggaggagg acccggcggc cgccttcctg gcccagcagg
      781 agagcgagat tgcaggcata gagaacgacg agggcttcgg ggcacctgcc ggcagccatg
      841 cggcccccgc acagccgggc cccacgagtg gggctggttc tgaggacatg gggaccacag
      901 tcaatggaga tgtgtttcag gaggccaacg gtcctgctga tggctacgca gccattgccc
      961 aggctgacag gctgacccag gagcctgaga gcatccgcaa gtggcgagag gagcagagga
     1021 aacggctgca agagctggat gctgcatcta aggtcacgga acaggaatgg cgggagaagg
     1081 ccaagaagga cctggaggag tggaaccagc gccagagtga acaagtagag aagaacaaga
     1141 tcaacaaccg gatcgctgac aaagcattct accagcagcc agatgctgat atcatcggct
     1201 acgtggcatc cgaggaggct ttcgtgaagg aatccaagga ggagacccca ggcacagagt
     1261 gggagaaggt ggcccagcta tgtgacttca accccaagag cagcaagcag tgcaaagatg
     1321 tgtcccgcct gcgctcggtg ctcatgtccc tgaagcagac gccactgtcc cgcggcggat
     1381 ccggagggtc tgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1441 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1501 agctgaccct gaagctcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1561 tgaccacctt cggctacggc ctgatgtgct tcgcgcgcta tccagaccac atgaagcagc
     1621 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1681 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1741 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1801 tcgagtacgc ttggcccgtg gtgaatgtct atatcatggc cgacaagcag aagaacggca
     1861 tcaaggtgaa cttctctatc tatcataaca tcgaggacgg cagcgtgcag ctcgccgacc
     1921 actaccagca gaacaccccc atcggcgacg ggcccgtgct gctgcccgac aaccactacc
     1981 tgagctactc ggctaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2041 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     2101 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2161 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2221 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2281 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2341 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2401 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2461 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2521 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2581 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2641 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2701 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2761 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     2821 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2881 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     2941 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3001 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3061 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3121 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3181 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3241 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     3301 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3361 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3421 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3481 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     3541 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3601 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3661 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3721 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3781 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3841 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3901 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3961 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4021 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4081 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4141 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4201 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4261 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     4321 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     4381 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4441 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4501 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4561 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4621 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4681 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4741 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4801 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4861 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4921 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4981 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5041 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5101 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5161 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5221 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5281 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5341 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5401 ttctgtggat aaccgtatta ccg
//
