LOCUS       sequence-207084-        5236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Halo-Tev-Flag-Ube2v1 (human) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5236)
  AUTHORS   Zhao Y, Long MJC, Wang Y, Zhang S, Aye Y
  TITLE     Ube2V2 Is a Rosetta Stone Bridging Redox and Ubiquitin Codes, 
            Coordinating DNA Damage Responses.
  JOURNAL   ACS Cent Sci. 2018 Feb 28;4(2):246-259. doi: 
            10.1021/acscentsci.7b00556. Epub 2018 Jan 17.
  PUBMED    29532025
REFERENCE   2  (bases 1 to 5236)
  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: 
            "10.1021/acscentsci.7b00556"; journalName: "ACS Cent Sci"; date: 
            "2018-02-28- 28"; volume: "4"; issue: "2"; pages: "246-259"
COMMENT     Sequence Label: Halo-Tev-Flag-Ube2v1
FEATURES             Location/Qualifiers
     source          1..5236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1957
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2006..2029
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2650..2668)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2705..2826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2716..2735)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2770..2789
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3073..3177
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3178..3246,3247..4038)
                     /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(3396..3415)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4196..4784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4276..4295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4900..5183
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcaaa gccaccatgg cagaaatcgg
     1081 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1141 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1201 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1261 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1321 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1381 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1441 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1501 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1561 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1621 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1681 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1741 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1801 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1861 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1921 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccactg aggatctgta
     1981 ctttcagagc gataacgcga tcgccgacta caaggatgac gacgataaga tgccaggaga
     2041 ggttcaagcg tcttacctga agtcacaaag caaactgagt gatgaaggaa gacttgaacc
     2101 tagaaaattt cactgcaaag gagtaaaagt ccctcgcaat ttccgactgt tggaagaact
     2161 cgaagaaggc cagaaaggag taggagatgg cacagttagc tggggtctag aagatgacga
     2221 agacatgaca cttacaagat ggacagggat gataattggg cctccaagaa caatttatga
     2281 aaaccgaata tatagcctta aaatagaatg tggacctaaa tacccagaag cacccccctt
     2341 tgtaagattt gtaacaaaaa ttaatatgaa tggagtaaat agttctaatg gagtggtgga
     2401 cccaagagcc atatcagtgc tagcaaaatg gcagaattca tatagcatca aagttgtcct
     2461 gcaagagctt cggcgcctaa tgatgtctaa agaaaatatg aaactccctc agccgcccga
     2521 aggacagtgt tacagcaatt aaacgaattc gggctcggta cccggggatc ctctagagtc
     2581 gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     2641 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct tcgagcagac
     2701 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2761 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2821 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     2881 gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaattcta atggatcctc
     2941 tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag
     3001 caccgtttct gcggactggc tttctacgta atggtttctt agacgtcagg tggcactttt
     3061 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3121 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3181 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3241 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3301 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3361 gaacgttttc caatgatgag cactttcaaa gttctgctat gtggcgcggt attatcccgt
     3421 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3481 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3541 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aactatcgga
     3601 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3661 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3721 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3781 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3841 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3901 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3961 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4021 ctgattaagc attggtaatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc
     4081 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4141 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4201 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4261 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4321 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4381 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4441 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4501 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4561 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4621 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4681 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4741 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct
     4801 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4861 gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaaataca ggaacgcacg
     4921 ctggatggcc cttcgctggg atggtgaaac catgaaaaat ggcagcttca gtggattaag
     4981 tgggggtaat gtggcctgta ccctctggtt gcataggtat tcatacggtt aaaatttatc
     5041 aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt
     5101 gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg gattatggta aatccactta
     5161 ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat gcgtccggcg
     5221 tagaggatcg agatct
//
