LOCUS       sequence-379896-        7473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human W279A long-form UBASH3A with V5 tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7473)
  TITLE     Concannon Lab UBASH3A Plasmids 2022
REFERENCE   2  (bases 1 to 7473)
  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"
COMMENT     Sequence Label: V5-W279A-long-UBASH3A
FEATURES             Location/Qualifiers
     source          1..7473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        146..525
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        526..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     774..793
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        774..792
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             825..866
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             867..887
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    918..942
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2932..2956)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2975..2992)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2981..3205
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3251..3679
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3338..3357)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3548..3569
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3688..3708)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3693..4022
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3873..4008
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3935..3954
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4089..4883
                     /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(4143..4162)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4753..4772
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5057..5178
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5094..5113)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5148..5167
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5227..5243)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5227..5243)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5240..5262)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5251..5267
                     /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(5275..5281,5282..5299,5300..5305))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5320..5341
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5458..5475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5629..6217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5709..5728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6388..7179,7180..7248))
                     /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     7011..7030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7249..7353)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7428..7447)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
       61 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
      121 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
      181 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      241 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      301 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      361 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      421 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      481 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      541 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      601 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      661 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
      781 actcactata gggagaccca agctggctag ttaagcttac catgggtaag cctatcccta
      841 accctctcct cggtctcgat tctacggaaa acctgtattt tcagggcccg gatccgtcga
      901 cgaattctgc agatatcaca agtttgtaca aaaaagcagg cttcatggca gcgggggaga
      961 cgcagctcta cgccaaggtc tccaacaagc tcaagagccg cagcagcccc tcgctcctgg
     1021 agcccctcct ggccatgggc ttcccggtgc acaccgcgct gaaagcgttg gcagccacgg
     1081 ggaggaagac ggcggaggag gccttggcct ggctgcatga tcattgcaat gacccttccc
     1141 tagacgaccc catcccccag gagtatgccc ttttcctctg tccaacgggg cccctgctgg
     1201 aaaaacttca agagttctgg agagagagca agcgccagtg tgcaaagaac agagctcatg
     1261 aggtcttccc acacgtgaca ctctgtgact tcttcacgtg tgaagaccag aaggtggaat
     1321 gcctgtacga ggcgctgaag agagctggag acaggctcct gggctccttc cccacggccg
     1381 tgcctctggc tctccactcc tccatcagct acctcggctt cttcgtcagt ggcagccccg
     1441 cagacgtcat ccgggaattc gccatgacct tcgccacgga agcatctctc ttagcaggca
     1501 cttccgtttc ccgcttctgg attttcagcc aggtgcctgg acatggccct aacctgaggc
     1561 tgagcaattt aactagagcc tccttcgtga gccactacat ccttcaaaaa tactgctccg
     1621 tgaagccttg caccaaacag ctgcatctga ccttggccca caagttctac ccccaccacc
     1681 agaggacgct ggagcagctg gccagagcca tccccctggg ccacagctgc cagtggaccg
     1741 cagcactcta ctcccgagac atgcgctttg tgcactacca gaccctgaga gccctattcc
     1801 agtacaaacc ccagaacgtg gatgagctga cgctaagtcc tggtgactac atctttgtgg
     1861 accccacgca gcaggacgaa gccagcgagg gcgcggtgat tgggatctca cagcggacgg
     1921 gctgccgggg cttcctgccg gaaaactaca cggatcgagc cagtgagtct gacacgtggg
     1981 tgaagcacag gatgtacacc ttcagtctag ccacagacct gaactccaga aaggatggtg
     2041 aagccagcag cagatgcagc ggggaatttc ttccacaaac ggcaaggagt cttagcagct
     2101 tacaggcctt gcaggctacc gttgcaagga agagtgtgct ggtggttcgc cacggggaga
     2161 gagtggatca gatcttcggg aaggcatggc tgcagcaatg ctccactcct gatgggaaat
     2221 actacaggcc agacctgaat ttcccctgca gtctgcccag gcggagtcgt gggatcaaag
     2281 actttgaaaa cgatccccca ttatcatcgt gtggcatttt ccagtccaga attgcagggg
     2341 acgcgctact ggacagtggt atcagaatca gctctgtgtt tgcctcccca gccctccgct
     2401 gtgtgcagac ggccaaactc atcctggaag aactcaaact ggagaaaaaa atcaagatac
     2461 gagtggaacc tggaatcttt gaatggacaa aatgggaagc tggcaaaacc accccaaccc
     2521 tcatgagcct ggaagagctg aaagaggcaa atttcaacat tgacactgat tacaggcccg
     2581 cgtttcccct gtccgccctc atgccggccg agagctacca ggagtacatg gacaggtgca
     2641 cggcgagcat ggtgcaaatc gtcaacacct gcccgcagga cacgggtgtc atcctaattg
     2701 tgagtcacgg ctccactctg gactcctgca cgcggccact gctcgggctg ccgccccggg
     2761 aatgtgggga ttttgcccaa ctcgtgagaa agatcccttc cctgggcatg tgcttctgtg
     2821 aagaaaataa agaggaagga aaatgggagt tggtgaaccc accggtgaag accctgaccc
     2881 acggggcgaa cgcagcattt aactggagga actggatctc aggcaactga aacccagctt
     2941 tcttgtacaa agtggtgata aacccgctga tcagcctcga ctgtgccttc tagttgccag
     3001 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3061 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3121 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3181 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3241 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3301 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3361 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3421 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3481 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3541 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3601 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3661 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3721 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3781 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3841 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3901 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3961 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4021 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4081 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4141 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4201 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4261 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4321 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4381 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4441 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4501 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4561 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4621 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4681 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4741 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4801 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4861 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4921 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4981 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5041 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5101 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5161 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5221 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5281 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5341 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5401 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5461 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6061 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7381 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     7441 gatgccgcat agttaagcca gtatctgctc cct
//
