LOCUS       sequence-361319-        7883 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc-USP10 construct in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7883)
  AUTHORS   Jia R, Bonifacino JS
  TITLE     The ubiquitin isopeptidase USP10 deubiquitinates LC3B to increase 
            LC3B levels and autophagic activity.
  JOURNAL   J Biol Chem. 2021 Jan-Jun;296:100405. doi: 
            10.1016/j.jbc.2021.100405. Epub 2021 Feb 10.
  PUBMED    33577797
REFERENCE   2  (bases 1 to 7883)
  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: "J Biol 
            Chem. 2021 Jan-Jun;296:100405. doi: 10.1016/j.jbc.2021.100405. Epub 
            2021 Feb 10."
COMMENT     Sequence Label: pCI-myc-USP10
FEATURES             Location/Qualifiers
     source          1..7883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        64..443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        444..655
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     606..626
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     652..676
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          816..948
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     931..950
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        993..1011
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1026..1055
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3513..3634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3524..3543)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3578..3597
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3820..4275
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3907..3926)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4117..4138
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4392..4749
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4392..4583
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4415..4435)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4600..4735
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4800..5594
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4854..4873)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5464..5483
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5658..5706
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5747..5766)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5866..5888
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5926..5944)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6012..6116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6117..6185,6186..6977)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6335..6354)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7148..7736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7637..7656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgccat
       61 gttggcattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      121 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      181 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      241 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      301 atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg
      361 cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg
      421 tattagtcat cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat
      481 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      541 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactgcgat cgcccgcccc
      601 gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt
      661 agtgaaccgt cagatcacta gaagctttat tgcggtagtt tatcacagtt aaattgctaa
      721 cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagtgact ctcttaaggt
      781 agccttgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg
      841 tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat
      901 aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt gtccactccc
      961 agttcaatta cagctcttaa ggctagagta cttaatacga ctcactatag gctagcctcg
     1021 agatggaaca aaaactcatc tcagaggagg atctcggaag cggatccgcc ctccacagcc
     1081 cgcagtatat ttttggagat tttagccctg atgaattcaa tcaattcttt gtgactcctc
     1141 gatcttcagt tgagcttcct ccatacagtg gaacagttct gtgtggcaca caggctgtgg
     1201 ataaactacc tgatggacaa gaatatcaga gaattgagtt tggtgtcgat gaagtcattg
     1261 aacccagtga cactttgccg agaaccccca gctacagtat ttcaagcaca ctgaaccctc
     1321 aggcccctga atttattctc ggttgtacag cttccaaaat aacccctgat ggtatcacta
     1381 aagaagcaag ctatggctcc atcgactgcc agtacccagg ctctgccctc gctttggatg
     1441 gaagttctaa tgtggaggcg gaagttttgg aaaatgatgg tgtctcaggt ggtcttggac
     1501 aaagggagcg taaaaagaag aaaaagcggc cacctggata ttacagctat ttgaaagatg
     1561 gtggcgatga tagtatctcc acagaagccc tggtcaatgg ccatgccaat tcagcagtcc
     1621 cgaacagtgt cagtgcagag gatgcagaat ttatgggtga catgcctccg ccacttacgc
     1681 ccaggacttg taacagcccc cagaactcca cagactctgt cagtgacatt gtgcctgaca
     1741 gtcctttccc cggagcactc ggcagtgaca ccaggactgc agggcagcca gaggggggcc
     1801 ccggggctga ttttggtcag tcctgcttcc ctgcagaggc tggcagagac accctgtcaa
     1861 ggacagctgg ggctcagccc tgcgttggta ccgatactac tgaaaacctt ggagttgcta
     1921 atggacaaat acttgaatcc tcgggtgagg gcacagctac caacggggtg gagttgcaca
     1981 ccacggaaag catagacttg gacccaacca aacccgagag tgcatcacct cctgctgacg
     2041 gcacgggctc tgcatcaggc acccttcctg tcagccagcc caagtcctgg gccagcctct
     2101 ttcatgattc taagccctct tcctcctcgc cggtggccta tgtggaaact aagtattccc
     2161 ctcccgccat atctcccctg gtttctgaaa agcaggttga agtcaaagaa gggcttgttc
     2221 cggtttcaga ggatcctgta gccataaaga ttgcagagtt gctggagaat gtaaccctaa
     2281 tccataaacc agtgtcgttg caaccccgtg ggctgatcaa taaagggaac tggtgctaca
     2341 ttaatgctac actgcaggca ttggttgctt gcccgccgat gtaccacctg atgaagttca
     2401 ttcctctgta ttccaaagtg caaaggcctt gtacgtcaac acccatgata gacagctttg
     2461 ttcggctaat gaatgagttc actaatatgc cagtacctcc aaaaccccga caagctcttg
     2521 gagataaaat cgtgagggat attcgccctg gagctgcctt tgagcccaca tatatttaca
     2581 gactcctgac agttaacaag tcaagcctgt ctgaaaaggg tcgacaagaa gatgctgagg
     2641 aatacttagg cttcattcta aatggacttc atgaggaaat gttgaaccta aagaagcttc
     2701 tctcaccaag taatgaaaaa cttacgattt ccaacggccc caaaaaccac tcggtcaatg
     2761 aagaagagca ggaagaacaa ggtgaaggaa gcgaggatga atgggaacaa gtgggccccc
     2821 ggaacaagac ttccgtcacc cgccaggcgg attttgttca gactccaatc accggcattt
     2881 ttggtggaca catcaggtct gtggtttacc agcagagttc aaaagaatct gccactttgc
     2941 agccattttt cacgttgcag ttggatatcc agtcagacaa gatacgcaca gtccaggatg
     3001 cactggagag cttggtggca agagaatctg tccaaggtta taccacaaaa accaaacaag
     3061 aggttgagat aagtcgaaga gtgactctgg aaaaactccc tcctgtcctc gtgctgcacc
     3121 tgaaacgatt cgtttatgag aagactggtg ggtgccagaa gcttatcaaa aatattgaat
     3181 atcctgtgga cttggaaatt agtaaagaac tgctttctcc aggggttaaa aataagaatt
     3241 ttaaatgcca ccgaacctat cggctctttg cagtggtcta ccatcacggc aacagtgcga
     3301 cgggcggcca ttacactaca gacgtcttcc agatcggtct gaatggctgg ctgcgcatcg
     3361 atgaccagac agtcaaggtg atcaaccagt accaggtggt gaaaccaact gctgaacgca
     3421 cagcctacct cctgtattac cgccgagtgg acctgctgta acccgggcgg ccgcttccct
     3481 ttagtgaggg ttaatgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     3541 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3601 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3661 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3721 tggtaaaatc cgataaggat cgatccgggc tggcgtaata gcgaagaggc ccgcaccgat
     3781 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat
     3841 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3901 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3961 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4021 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4081 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4141 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4201 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4261 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4321 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     4381 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     4441 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4501 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4561 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4621 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4681 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4741 ttttgcaaaa agcttgattc ttctgacaca acagtctcga acttaaggct agagccacca
     4801 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4861 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4921 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4981 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5041 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5101 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5161 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5221 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5281 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     5341 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5401 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5461 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5521 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5581 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5641 gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     5701 tgtgtgaatc gatagcgata aggatccgcg tatggtgcac tctcagtaca atctgctctg
     5761 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5821 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5881 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5941 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6001 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6061 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6121 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6181 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6241 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6301 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6361 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6421 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6481 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6541 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6601 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6661 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6721 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6781 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6841 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6901 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6961 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7021 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7081 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7141 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7201 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7261 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7321 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7381 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7441 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7501 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7561 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7621 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7681 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7741 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatggctc
     7801 gacagatctt caatattggc cattagccat attattcatt ggttatatag cataaatcaa
     7861 tattggctat tggccattgc ata
//
