LOCUS       sequence-379910-        7667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human wild-type 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: WT-short-UBASH3A-V5-6xHis
FEATURES             Location/Qualifiers
     source          1..7667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        242..1420
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1411
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1368..1388
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1437..1456
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1437..1455
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1487..1511
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3384..3408)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3434..3475
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3485..3502
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3525..3542)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3531..3755
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3801..4229
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3888..3907)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4098..4119
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4238..4258)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4243..4572
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4423..4558
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4485..4504
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4620..4667
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4686..5084
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5242..5363
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5279..5298)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5333..5352
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5412..5428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5412..5428)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5425..5447)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5436..5452
                     /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(5460..5466,5467..5484,5485..5490))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5505..5526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5643..5660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5803..6391)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5883..5902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6562..7353,7354..7422))
                     /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     7185..7204
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7423..7527)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7602..7621)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
       61 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      121 gcgttgacat tgattattga ctaggctttt gcaaaaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
      601 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
      661 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
      721 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
      781 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
      841 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
      901 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
      961 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     1021 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     1081 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     1141 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     1201 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     1261 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     1321 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     1381 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggaattagct tggtactaat
     1441 acgactcact atagggagac ccaagctggc taggtaagct tgatcaacaa gtttgtacaa
     1501 aaaagcaggc ttcatggcag cgggggagac gcagctctac gccaaggtct ccaacaagct
     1561 caagagccgc agcagcccct cgctcctgga gcccctcctg gccatgggct tcccggtgca
     1621 caccgcgctg aaagcgttgg cagccacggg gaggaagacg gcggaggagg ccttggcctg
     1681 gctgcatgat cattgcaatg acccttccct agacgacccc atcccccagg agtatgccct
     1741 tttcctctgt ccaacggggc ccctgctgga aaaacttcaa gagttctgga gagagagcaa
     1801 gcgccagtgt gcaaagaaca gagctcatga ggtcttccca cacgtgacac tctgtgactt
     1861 cttcacgtgt gaagaccaga aggtggaatg cctgtacgag gcgctgaaga gagctggaga
     1921 caggctcctg ggctccttcc ccacggccgt gcctctggct ctccactcct ccatcagcta
     1981 cctcggcttc ttcgtcagtg gcagccccgc agacgtcatc cgggaattcg ccatgacctt
     2041 cgccacggaa gcatctctct tagcagactg ctccgtgaag ccttgcacca aacagctgca
     2101 tctgaccttg gcccacaagt tctaccccca ccaccagagg acgctggagc agctggccag
     2161 agccatcccc ctgggccaca gctgccagtg gaccgcagca ctctactccc gagacatgcg
     2221 ctttgtgcac taccagaccc tgagagccct attccagtac aaaccccaga acgtggatga
     2281 gctgacgcta agtcctggtg actacatctt tgtggacccc acgcagcagg acgaagccag
     2341 cgagggctgg gtgattggga tctcacagcg gacgggctgc cggggcttcc tgccggaaaa
     2401 ctacacggat cgagccagtg agtctgacac gtgggtgaag cacaggatgt acaccttcag
     2461 tctagccaca gacctgaact ccagaaagga tggtgaagcc agcagcagat gcagcgggga
     2521 atttcttcca caaacggcaa ggagtcttag cagcttacag gccttgcagg ctaccgttgc
     2581 aaggaagagt gtgctggtgg ttcgccacgg ggagagagtg gatcagatct tcgggaaggc
     2641 atggctgcag caatgctcca ctcctgatgg gaaatactac aggccagacc tgaatttccc
     2701 ctgcagtctg cccagacgga gtcgtgggat caaagacttt gaaaacgatc ccccattatc
     2761 atcgtgtggc attttccagt ccagaattgc aggggacgcg ctactggaca gtggtatcag
     2821 aatcagctct gtgtttgcct ccccagccct ccgctgtgtg cagacggcca aactcatcct
     2881 ggaagaactc aaactggaga aaaaaatcaa gatacgagtg gaacctggaa tctttgaatg
     2941 gacaaaatgg gaagctggca agaccacccc aaccctcatg agcctggaag agctgaaaga
     3001 ggcaaatttc aacattgaca ctgattacag gcccgcgttt cccctgtccg ccctcatgcc
     3061 ggccgagagc taccaggagt acatggacag gtgcacggcg agcatggtgc aaatcgtcaa
     3121 cacctgtccg caggacacgg gtgtcatcct aattgtgagt cacggctcca ctctggactc
     3181 ctgcacgcgg ccactgctcg ggctgccgcc ccgggaatgt ggggattttg cccaactcgt
     3241 gagaaagatc ccttccctgg gcatgtgctt ctgtgaagaa aataaagagg aaggaaaatg
     3301 ggagttggtg aacccaccgg tgaagaccct gacccacggg gcgaacgcag catttaactg
     3361 gaggaactgg atctcaggca acaacccagc tttcttgtac aaagtggttg atctagaggg
     3421 cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     3481 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     3541 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3601 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3661 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3721 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3781 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3841 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3901 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3961 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4021 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4081 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4141 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4201 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4261 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4321 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4381 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4441 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4501 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4561 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     4621 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     4681 aaaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
     4741 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
     4801 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
     4861 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
     4921 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg
     4981 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
     5041 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagcactt cgtggccgag
     5101 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5161 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5221 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5281 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5341 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5401 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5461 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5521 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5581 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5641 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5701 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5761 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5821 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5881 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5941 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6001 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6061 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6121 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6181 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6241 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6301 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6361 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6421 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6481 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6541 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6601 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6661 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6721 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6781 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6841 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6901 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6961 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7021 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7081 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7141 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7201 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7261 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7321 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7381 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7441 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7501 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7561 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     7621 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgc
//
