LOCUS       sequence-494421-        5539 bp    DNA     circular SYN 02-DEC-2025
DEFINITION  Mammalian expression of human CHIP with U-box domain deleted, and a 
            myc-tag on its C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5539)
  AUTHORS   Lee J, Chin N, Zou J, Mazli WNAB, Jarnik M, Saidi L, Xu Y, Jeong E, 
            Suh J, Replogle J, Ward ME, Bonifacino JS, Zheng W, Hao L, Ye Y
  TITLE     CHIP protects lysosomes from CLN4 mutant-induced membrane damage.
  JOURNAL   Nat Cell Biol. 2025 Aug 25. doi: 10.1038/s41556-025-01738-2.
  PUBMED    40855364
REFERENCE   2  (bases 1 to 5539)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Cell 
            Biol. 2025 Aug 25. doi: 10.1038/s41556-025-01738-2."
COMMENT     Sequence Label: pCMV6 CHIP delU-box-Myc
FEATURES             Location/Qualifiers
     source          1..5539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(93..887)
                     /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             complement(93..887)
                     /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(204..223)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     814..833
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(922..1279)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      936..1071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1088..1279
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1236..1256
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(1281..1385)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1412..1867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1499..1518)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1709..1730
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1993..2015
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2007..2024
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2008..2024
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2185..2564
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2565..2768
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2719..2739
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2794..2813
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2794..2812
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3573..3602
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3638..3654)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3638..3654)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3665..4287
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3864..3883)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(4436..5024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4516..4535)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5353..5400
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5425..5444
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
ORIGIN
        1 caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc
       61 ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
      121 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
      181 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcga
      241 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
      301 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc
      361 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
      421 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
      481 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
      541 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
      601 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
      661 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg
      721 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
      781 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
      841 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
      901 catcctgtct cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
      961 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     1021 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     1081 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     1141 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     1201 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     1261 ctggttcttt ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt
     1321 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     1381 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     1441 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1501 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1561 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1621 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     1681 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1741 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     1801 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     1861 acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     1921 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     1981 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctgatctata
     2041 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     2101 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     2161 catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
     2221 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     2281 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2341 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2401 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg
     2461 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc
     2521 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2581 agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2641 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2701 ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2761 gcagagctcg tttagtgaac cgtcagaatt ttgtaatacg actcactata gggcggccgg
     2821 gaattcgtcg actggatccg gtaccgagga gatctgccgc cgcgatcgcc atgaagggca
     2881 aggaggagaa ggagggcggc gcacggctgg gcgctggcgg cggaagcccc gagaagagcc
     2941 cgagcgcgca ggagctcaag gagcagggca atcgtctgtt cgtgggccga aagtacccgg
     3001 aggcggcggc ctgctacggc cgcgcgatca cccggaaccc gctggtggcc gtgtattaca
     3061 ccaaccgggc cttgtgctac ctgaagatgc agcagcacga gcaggccctg gccgactgcc
     3121 ggcgcgccct ggagctggac gggcagtctg tgaaggcgca cttcttcctg gggcagtgcc
     3181 agctggagat ggagagctat gatgaggcca tcgccaatct gcagcgagct tacagcctgg
     3241 ccaaggagca gcggctgaac ttcggggacg acatccccag cgctcttcga atcgcgaaga
     3301 agaagcgctg gaacagcatt gaggagcggc gcatccacca ggagagcgag ctgcactcct
     3361 acctctccag gctcattgcc gcggagcgtg agagggagct ggaagagtgc cagcgaaacc
     3421 acgagggtga tgaggacgac agccacgtcc gggcccagca ggcctgcatt gaggccaagc
     3481 acgacaagta catggcggac atggacgagc ttttttctca ggtggatgag aagaggaaga
     3541 agcgaagcgg accgacgcgt acgcggccgc tcgagcagaa actcatctca gaagaggatc
     3601 tgcagatcct cttctgagat gaccgcggcc ggccgcggtc atagctgttt cctgaacaga
     3661 tcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     3721 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     3781 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     3841 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     3901 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     3961 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     4021 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     4081 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     4141 tctgatttta aaataactat accagcagga ggacgtccag acacagcata ggctacctgg
     4201 ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg
     4261 gtccactcag tagatgcctg ttgaattggg tacgcggcca gcggcgagcg gtatcagctc
     4321 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4381 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4441 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4501 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4561 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4621 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4681 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4741 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4801 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4861 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4921 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4981 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5041 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5101 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5161 tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg
     5221 cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg
     5281 cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa
     5341 ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg
     5401 tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag
     5461 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     5521 ggaaaacgat tccgaagcc
//
