LOCUS       sequence-379890-        7359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human K202R 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-K202R-short-UBASH3A
FEATURES             Location/Qualifiers
     source          1..7359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        83..462
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        463..666
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     617..637
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     711..730
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        711..729
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             762..803
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             804..824
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    855..879
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2755..2779)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2798..2815)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2804..3028
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3074..3502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3161..3180)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3371..3392
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3511..3531)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3516..3845
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3696..3831
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3758..3777
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3912..4706
                     /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(3966..3985)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4576..4595
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4880..5001
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4917..4936)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4971..4990
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5050..5066)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5050..5066)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5063..5085)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5074..5090
                     /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(5098..5104,5105..5122,5123..5128))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5143..5164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5281..5298)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5452..6040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5532..5551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6211..7002,7003..7071))
                     /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     6834..6853
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7072..7176)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7251..7270)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
       61 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      121 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      181 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      241 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      301 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      361 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      421 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      481 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      541 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      601 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      661 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
      721 cactataggg agacccaagc tggctagtta agcttaccat gggtaagcct atccctaacc
      781 ctctcctcgg tctcgattct acggaaaacc tgtattttca gggcccggat ccgtcgacga
      841 attctgcaga tatcacaagt ttgtacaaaa aagcaggctt catggcagcg ggggagacgc
      901 agctctacgc caaggtctcc aacaagctca agagccgcag cagcccctcg ctcctggagc
      961 ccctcctggc catgggcttc ccggtgcaca ccgcgctgaa agcgttggca gccacgggga
     1021 ggaagacggc ggaggaggcc ttggcctggc tgcatgatca ttgcaatgac ccttccctag
     1081 acgaccccat cccccaggag tatgcccttt tcctctgtcc aacggggccc ctgctggaaa
     1141 aacttcaaga gttctggaga gagagcaagc gccagtgtgc aaagaacaga gctcatgagg
     1201 tcttcccaca cgtgacactc tgtgacttct tcacgtgtga agaccagaag gtggaatgcc
     1261 tgtacgaggc gctgaagaga gctggagaca ggctcctggg ctccttcccc acggccgtgc
     1321 ctctggctct ccactcctcc atcagctacc tcggcttctt cgtcagtggc agccccgcag
     1381 acgtcatccg ggaattcgcc atgaccttcg ccacggaagc atctctctta gcagactgct
     1441 ccgtgaagcc ttgcaccaaa cagctgcatc tgaccttggc ccacaggttc tacccccacc
     1501 accagaggac gctggagcag ctggccagag ccatccccct gggccacagc tgccagtgga
     1561 ccgcagcact ctactcccga gacatgcgct ttgtgcacta ccagaccctg agagccctat
     1621 tccagtacaa accccagaac gtggatgagc tgacgctaag tcctggtgac tacatctttg
     1681 tggaccccac gcagcaggac gaagccagcg agggctgggt gattgggatc tcacagcgga
     1741 cgggctgccg gggcttcctg ccggaaaact acacggatcg agccagtgag tctgacacgt
     1801 gggtgaagca caggatgtac accttcagtc tagccacaga cctgaactcc agaaaggatg
     1861 gtgaagccag cagcagatgc agcggggaat ttcttccaca aacggcaagg agtcttagca
     1921 gcttacaggc cttgcaggct accgttgcaa ggaagagtgt gctggtggtt cgccacgggg
     1981 agagagtgga tcagatcttc gggaaggcat ggctgcagca atgctccact cctgatggga
     2041 aatactacag gccagacctg aatttcccct gcagtctgcc cagacggagt cgtgggatca
     2101 aagactttga aaacgatccc ccattatcat cgtgtggcat tttccagtcc agaattgcag
     2161 gggacgcgct actggacagt ggtatcagaa tcagctctgt gtttgcctcc ccagccctcc
     2221 gctgtgtgca gacggccaaa ctcatcctgg aagaactcaa actggagaaa aaaatcaaga
     2281 tacgagtgga acctggaatc tttgaatgga caaaatggga agctggcaag accaccccaa
     2341 ccctcatgag cctggaagag ctgaaagagg caaatttcaa cattgacact gattacaggc
     2401 ccgcgtttcc cctgtccgcc ctcatgccgg ccgagagcta ccaggagtac atggacaggt
     2461 gcacggcgag catggtgcaa atcgtcaaca cctgtccgca ggacacgggt gtcatcctaa
     2521 ttgtgagtca cggctccact ctggactcct gcacgcggcc actgctcggg ctgccgcccc
     2581 gggaatgtgg ggattttgcc caactcgtga gaaagatccc ttccctgggc atgtgcttct
     2641 gtgaagaaaa taaagaggaa ggaaaatggg agttggtgaa cccaccggtg aagaccctga
     2701 cccacggggc gaacgcagca tttaactgga ggaactggat ctcaggcaac tgaaacccag
     2761 ctttcttgta caaagtggtg ataaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2821 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2881 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2941 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3001 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3061 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3121 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3181 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3241 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3301 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3361 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3421 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3481 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3541 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3601 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3661 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3721 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3781 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3841 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3901 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3961 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4021 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4081 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4141 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4201 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4261 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4321 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4381 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4441 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4501 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4561 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4621 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4681 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4741 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4801 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4861 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4921 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4981 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5041 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5101 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5161 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5221 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5281 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5341 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5401 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5461 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5521 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5581 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5641 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5701 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5761 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5821 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5881 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5941 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6001 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6061 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6121 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6181 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6241 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6301 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6361 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6421 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6481 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6541 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6601 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6661 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6721 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6781 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6841 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6901 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6961 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7021 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7081 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7141 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7201 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     7261 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     7321 tagtgcgcga gcaaaattta agctacaaca aggcaaggc
//
