LOCUS       sequence-386307-        7667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human K202R short-form UBASH3A with V5-6xHis
            tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7667)
  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 7667)
  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: K202R-short-UBASH3A-V5-6xHis
FEATURES             Location/Qualifiers
     source          1..7667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(166..185)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        473..1651
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          704..1642
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1599..1619
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1668..1687
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1668..1686
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1718..1742
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3615..3639)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3665..3706
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3716..3733
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3756..3773)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3762..3986
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4032..4460
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4119..4138)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4329..4350
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4469..4489)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4474..4803
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4654..4789
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4716..4735
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4851..4898
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4917..5315
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5473..5594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5510..5529)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5564..5583
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5643..5659)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5643..5659)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5656..5678)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5667..5683
                     /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(5691..5697,5698..5715,5716..5721))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5736..5757
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5874..5891)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6034..6622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6114..6133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6793..7584,7585..7653))
                     /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     7416..7435
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7654..7667,1..91))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
      181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga
      421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg
      481 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
      541 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
      601 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
      661 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
      721 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
      781 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
      841 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
      901 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
      961 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     1021 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     1081 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     1141 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     1201 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     1261 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     1321 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     1381 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     1441 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     1501 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     1561 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     1621 aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac
     1681 tatagggaga cccaagctgg ctaggtaagc ttgatcaaca agtttgtaca aaaaagcagg
     1741 cttcatggca gcgggggaga cgcagctcta cgccaaggtc tccaacaagc tcaagagccg
     1801 cagcagcccc tcgctcctgg agcccctcct ggccatgggc ttcccggtgc acaccgcgct
     1861 gaaagcgttg gcagccacgg ggaggaagac ggcggaggag gccttggcct ggctgcatga
     1921 tcattgcaat gacccttccc tagacgaccc catcccccag gagtatgccc ttttcctctg
     1981 tccaacgggg cccctgctgg aaaaacttca agagttctgg agagagagca agcgccagtg
     2041 tgcaaagaac agagctcatg aggtcttccc acacgtgaca ctctgtgact tcttcacgtg
     2101 tgaagaccag aaggtggaat gcctgtacga ggcgctgaag agagctggag acaggctcct
     2161 gggctccttc cccacggccg tgcctctggc tctccactcc tccatcagct acctcggctt
     2221 cttcgtcagt ggcagccccg cagacgtcat ccgggaattc gccatgacct tcgccacgga
     2281 agcatctctc ttagcagact gctccgtgaa gccttgcacc aaacagctgc atctgacctt
     2341 ggcccaccgg ttctaccccc accaccagag gacgctggag cagctggcca gagccatccc
     2401 cctgggccac agctgccagt ggaccgcagc actctactcc cgagacatgc gctttgtgca
     2461 ctaccagacc ctgagagccc tattccagta caaaccccag aacgtggatg agctgacgct
     2521 aagtcctggt gactacatct ttgtggaccc cacgcagcag gacgaagcca gcgagggctg
     2581 ggtgattggg atctcacagc ggacgggctg ccggggcttc ctgccggaaa actacacgga
     2641 tcgagccagt gagtctgaca cgtgggtgaa gcacaggatg tacaccttca gtctagccac
     2701 agacctgaac tccagaaagg atggtgaagc cagcagcaga tgcagcgggg aatttcttcc
     2761 acaaacggca aggagtctta gcagcttaca ggccttgcag gctaccgttg caaggaagag
     2821 tgtgctggtg gttcgccacg gggagagagt ggatcagatc ttcgggaagg catggctgca
     2881 gcaatgctcc actcctgatg ggaaatacta caggccagac ctgaatttcc cctgcagtct
     2941 gcccagacgg agtcgtggga tcaaagactt tgaaaacgat cccccattat catcgtgtgg
     3001 cattttccag tccagaattg caggggacgc gctactggac agtggtatca gaatcagctc
     3061 tgtgtttgcc tccccagccc tccgctgtgt gcagacggcc aaactcatcc tggaagaact
     3121 caaactggag aaaaaaatca agatacgagt ggaacctgga atctttgaat ggacaaaatg
     3181 ggaagctggc aagaccaccc caaccctcat gagcctggaa gagctgaaag aggcaaattt
     3241 caacattgac actgattaca ggcccgcgtt tcccctgtcc gccctcatgc cggccgagag
     3301 ctaccaggag tacatggaca ggtgcacggc gagcatggtg caaatcgtca acacctgtcc
     3361 gcaggacacg ggtgtcatcc taattgtgag tcacggctcc actctggact cctgcacgcg
     3421 gccactgctc gggctgccgc cccgggaatg tggggatttt gcccaactcg tgagaaagat
     3481 cccttccctg ggcatgtgct tctgtgaaga aaataaagag gaaggaaaat gggagttggt
     3541 gaacccaccg gtgaagaccc tgacccacgg ggcgaacgca gcatttaact ggaggaactg
     3601 gatctcaggc aacaacccag ctttcttgta caaagtggtt gatctagagg gcccgcggtt
     3661 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     3721 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3781 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3841 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3901 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3961 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     4021 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4081 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4141 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4201 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4261 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4321 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4381 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4441 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4501 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4561 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4621 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4681 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4741 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4801 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
     4861 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     4921 ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca
     4981 gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca
     5041 tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc
     5101 tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg
     5161 agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc
     5221 ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg
     5281 aattgctgcc ctctggttat gtgtgggagg gctaagcact tcgtggccga ggagcaggac
     5341 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5401 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5461 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5521 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5581 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5641 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5701 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5761 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5821 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5881 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5941 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6001 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6061 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6121 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6181 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6241 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6301 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6361 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6421 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6481 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6541 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6601 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6661 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6721 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6781 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6841 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6901 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6961 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7021 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7081 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7141 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7201 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7261 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7321 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7381 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7441 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7501 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7561 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7621 aataagggcg acacggaaat gttgaatact catactcttc ctttttc
//
