LOCUS       sequence-388939-        5788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged ubiquitin in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5788)
  AUTHORS   Mattera R, Tsai YC, Weissman AM, Bonifacino JS
  TITLE     The Rab5 guanine nucleotide exchange factor Rabex-5 binds ubiquitin 
            (Ub) and functions as a Ub ligase through an atypical Ub-interacting
            motif and a zinc finger domain.
  JOURNAL   J Biol Chem. 2006 Mar 10;281(10):6874-83. doi: 
            10.1074/jbc.M509939200. Epub 2006 Jan 5.
  PUBMED    16407276
REFERENCE   2  (bases 1 to 5788)
  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"; doi: 
            "10.1074/jbc.M509939200"; journalName: "J Biol Chem"; date: 
            "2006-03-10- 10"; volume: "281"; issue: "10"; pages: "6874-83"
COMMENT     Sequence Label: pCI-neo-(HA)3-ubiquitin
FEATURES             Location/Qualifiers
     source          1..5788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        92..449
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        92..283
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(115..135)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      300..435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             500..1294
                     /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(554..573)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1164..1183
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1358..1406
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(1447..1466)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1566..1588
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1626..1644)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1712..1816
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1817..1885,1886..2677)
                     /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(2035..2054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2848..3436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3337..3356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3647..4026
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4027..4238
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4189..4209
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4235..4259
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          4399..4531
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4514..4533
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        4576..4594
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4618..4644
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4648..4674
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4681..4707
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4714..4941
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     polyA_signal    5001..5122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5012..5031)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5066..5085
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      5308..5763
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5395..5414)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5605..5626
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 catctgtgcg gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa
       61 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg
      121 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      181 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
      241 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
      301 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
      361 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
      421 gcttttttgg aggcctaggc ttttgcaaaa agcttgattc ttctgacaca acagtctcga
      481 acttaaggct agagccacca tgattgaaca agatggattg cacgcaggtt ctccggccgc
      541 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
      601 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
      661 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
      721 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
      781 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
      841 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
      901 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
      961 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     1021 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     1081 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     1141 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     1201 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     1261 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     1321 gaccaagcga cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt
     1381 acatctgtgt gttggttttt tgtgtgaatc gatagcgata aggatccgcg tatggtgcac
     1441 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     1501 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     1561 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     1621 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1681 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1741 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1801 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1861 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1921 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1981 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2041 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2101 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2161 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2221 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2281 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2341 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2401 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2461 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2521 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2581 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2641 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2701 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2761 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2821 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2881 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2941 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3001 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3061 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3121 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3181 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3241 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3301 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3361 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3421 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3481 gccttttgct cacatggctc gacagatctt caatattggc cattagccat attattcatt
     3541 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     3601 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttggca ttgattattg
     3661 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     3721 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     3781 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     3841 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     3901 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     3961 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     4021 accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg
     4081 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     4141 cgggactttc caaaatgtcg taacaactgc gatcgcccgc cccgttgacg caaatgggcg
     4201 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatca
     4261 ctagaagctt tattgcggta gtttatcaca gttaaattgc taacgcagtc agtgcttctg
     4321 acacaacagt ctcgaactta agctgcagtg actctcttaa ggtagccttg cagaagttgg
     4381 tcgtgaggca ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag
     4441 aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt
     4501 actgacatcc actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct
     4561 taaggctaga gtacttaata cgactcacta taggctagcc tcgaggccgc caccatgtac
     4621 ccgtacgatg ttcctgacta tgcgggctat ccctatgacg tcccggacta tgccatgggc
     4681 tacccttacg acgttccaga ttacgctgaa ttcatgcaga tcttcgtgaa gaccctgact
     4741 ggtaagacca tcaccctcga ggtggagccc agtgacacca tcgagaatgt caaggcaaag
     4801 atccaagata aggaaggcat ccctcctgat cagcagaggt tgatctttgc tgggaaacag
     4861 ctggaagatg gacgcaccct gtctgactac aacatccaga aagagtccac tctgcacttg
     4921 gtcctgcgct tgaggggggg ttaagtcgac ccgggcggcc gcttcccttt agtgagggtt
     4981 aatgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     5041 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     5101 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     5161 gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatccg
     5221 ataaggatcg atccgggctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     5281 cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta agcgcggcgg
     5341 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5401 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5461 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5521 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5581 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5641 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5701 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     5761 tttcctgatg cggtattttc tccttacg
//
