LOCUS       sequence-379897-        7359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human W279A short-form UBASH3A with V5 tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7359)
  AUTHORS   Ge Y, Paisie TK, Chen S, Concannon P
  TITLE     UBASH3A Regulates the Synthesis and Dynamics of TCR-CD3 Complexes.
  JOURNAL   J Immunol. 2019 Dec 1;203(11):2827-2836. doi: 
            10.4049/jimmunol.1801338. Epub 2019 Oct 28.
  PUBMED    31659016
REFERENCE   2  (bases 1 to 7359)
  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: 
            "10.4049/jimmunol.1801338"; journalName: "J Immunol"; date: 
            "2019-12-1- 1"; volume: "203"; issue: "11"; pages: "2827-2836"
COMMENT     Sequence Label: V5-W279A-short-UBASH3A
FEATURES             Location/Qualifiers
     source          1..7359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        149..528
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        529..732
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     683..703
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     777..796
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        777..795
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             828..869
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             870..890
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    921..945
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2821..2845)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2864..2881)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2870..3094
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3140..3568
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3227..3246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3437..3458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3577..3597)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3582..3911
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3762..3897
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3824..3843
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3978..4772
                     /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(4032..4051)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4642..4661
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4946..5067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4983..5002)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5037..5056
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5116..5132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5116..5132)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5129..5151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5140..5156
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5164..5170,5171..5188,5189..5194))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5209..5230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5347..5364)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5518..6106)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5598..5617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6277..7068,7069..7137))
                     /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     6900..6919
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7138..7242)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7317..7336)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
       61 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
      121 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
      181 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      241 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      301 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      361 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      421 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      481 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      541 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      601 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      661 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      721 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat
      781 acgactcact atagggagac ccaagctggc tagttaagct taccatgggt aagcctatcc
      841 ctaaccctct cctcggtctc gattctacgg aaaacctgta ttttcagggc ccggatccgt
      901 cgacgaattc tgcagatatc acaagtttgt acaaaaaagc aggcttcatg gcagcggggg
      961 agacgcagct ctacgccaag gtctccaaca agctcaagag ccgcagcagc ccctcgctcc
     1021 tggagcccct cctggccatg ggcttcccgg tgcacaccgc gctgaaagcg ttggcagcca
     1081 cggggaggaa gacggcggag gaggccttgg cctggctgca tgatcattgc aatgaccctt
     1141 ccctagacga ccccatcccc caggagtatg cccttttcct ctgtccaacg gggcccctgc
     1201 tggaaaaact tcaagagttc tggagagaga gcaagcgcca gtgtgcaaag aacagagctc
     1261 atgaggtctt cccacacgtg acactctgtg acttcttcac gtgtgaagac cagaaggtgg
     1321 aatgcctgta cgaggcgctg aagagagctg gagacaggct cctgggctcc ttccccacgg
     1381 ccgtgcctct ggctctccac tcctccatca gctacctcgg cttcttcgtc agtggcagcc
     1441 ccgcagacgt catccgggaa ttcgccatga ccttcgccac ggaagcatct ctcttagcag
     1501 actgctccgt gaagccttgc accaaacagc tgcatctgac cttggcccac aagttctacc
     1561 cccaccacca gaggacgctg gagcagctgg ccagagccat ccccctgggc cacagctgcc
     1621 agtggaccgc agcactctac tcccgagaca tgcgctttgt gcactaccag accctgagag
     1681 ccctattcca gtacaaaccc cagaacgtgg atgagctgac gctaagtcct ggtgactaca
     1741 tctttgtgga ccccacgcag caggacgaag ccagcgaggg cgcggtgatt gggatctcac
     1801 agcggacggg ctgccggggc ttcctgccgg aaaactacac ggatcgagcc agtgagtctg
     1861 acacgtgggt gaagcacagg atgtacacct tcagtctagc cacagacctg aactccagaa
     1921 aggatggtga agccagcagc agatgcagcg gggaatttct tccacaaacg gcaaggagtc
     1981 ttagcagctt acaggccttg caggctaccg ttgcaaggaa gagtgtgctg gtggttcgcc
     2041 acggggagag agtggatcag atcttcggga aggcatggct gcagcaatgc tccactcctg
     2101 atgggaaata ctacaggcca gacctgaatt tcccctgcag tctgcccaga cggagtcgtg
     2161 ggatcaaaga ctttgaaaac gatcccccat tatcatcgtg tggcattttc cagtccagaa
     2221 ttgcagggga cgcgctactg gacagtggta tcagaatcag ctctgtgttt gcctccccag
     2281 ccctccgctg tgtgcagacg gccaaactca tcctggaaga actcaaactg gagaaaaaaa
     2341 tcaagatacg agtggaacct ggaatctttg aatggacaaa atgggaagct ggcaagacca
     2401 ccccaaccct catgagcctg gaagagctga aagaggcaaa tttcaacatt gacactgatt
     2461 acaggcccgc gtttcccctg tccgccctca tgccggccga gagctaccag gagtacatgg
     2521 acaggtgcac ggcgagcatg gtgcaaatcg tcaacacctg tccgcaggac acgggtgtca
     2581 tcctaattgt gagtcacggc tccactctgg actcctgcac gcggccactg ctcgggctgc
     2641 cgccccggga atgtggggat tttgcccaac tcgtgagaaa gatcccttcc ctgggcatgt
     2701 gcttctgtga agaaaataaa gaggaaggaa aatgggagtt ggtgaaccca ccggtgaaga
     2761 ccctgaccca cggggcgaac gcagcattta actggaggaa ctggatctca ggcaactgaa
     2821 acccagcttt cttgtacaaa gtggtgataa acccgctgat cagcctcgac tgtgccttct
     2881 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2941 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3001 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3061 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3121 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3181 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3241 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3301 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3361 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3421 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3481 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3541 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3601 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3661 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3721 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3781 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3841 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3901 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3961 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4021 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4081 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4141 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4201 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4261 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4321 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4381 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4441 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4501 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4561 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4621 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4681 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4741 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4801 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4861 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4921 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4981 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5041 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5101 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5161 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5221 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5281 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5341 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5401 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5461 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5521 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5581 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5641 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5701 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5761 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5821 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5881 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5941 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6001 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6061 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6121 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6181 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6241 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6301 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6361 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6421 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6481 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6541 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6601 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6661 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6721 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6781 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6841 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6901 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6961 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7021 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7081 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7141 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7201 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7261 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     7321 atctgctctg atgccgcata gttaagccag tatctgctc
//
