LOCUS       sequence-386312-        7359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human K40R short-form UBASH3A with V5 tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7359)
  TITLE     Concannon Lab Plasmids
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"
COMMENT     Sequence Label: V5-K40R-short-UBASH3A
FEATURES             Location/Qualifiers
     source          1..7359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             914..955
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             956..976
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    1007..1031
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2907..2931)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2950..2967)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2956..3180
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3226..3654
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3313..3332)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3523..3544
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3663..3683)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3668..3997
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3848..3983
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3910..3929
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4064..4858
                     /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(4118..4137)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4728..4747
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5032..5153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5069..5088)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5123..5142
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5202..5218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5202..5218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5215..5237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5226..5242
                     /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(5250..5256,5257..5274,5275..5280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5295..5316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5433..5450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5604..6192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5684..5703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6363..7154,7155..7223))
                     /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     6986..7005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7224..7328)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttacc atgggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacggaaaa
      961 cctgtatttt cagggcccgg atccgtcgac gaattctgca gatatcacaa gtttgtacaa
     1021 aaaagcaggc ttcatggcag cgggggagac gcagctctac gccaaggtct ccaacaagct
     1081 caagagccgc agcagcccct cgctcctgga gcccctcctg gccatgggct tcccggtgca
     1141 caccgcgctg cgagcgttgg cagccacggg gaggaagacg gcggaggagg ccttggcctg
     1201 gctgcatgat cattgcaatg acccttccct agacgacccc atcccccagg agtatgccct
     1261 tttcctctgt ccaacggggc ccctgctgga aaaacttcaa gagttctgga gagagagcaa
     1321 gcgccagtgt gcaaagaaca gagctcatga ggtcttccca cacgtgacac tctgtgactt
     1381 cttcacgtgt gaagaccaga aggtggaatg cctgtacgag gcgctgaaga gagctggaga
     1441 caggctcctg ggctccttcc ccacggccgt gcctctggct ctccactcct ccatcagcta
     1501 cctcggcttc ttcgtcagtg gcagccccgc agacgtcatc cgggaattcg ccatgacctt
     1561 cgccacggaa gcatctctct tagcagactg ctccgtgaag ccttgcacca aacagctgca
     1621 tctgaccttg gcccacaagt tctaccccca ccaccagagg acgctggagc agctggccag
     1681 agccatcccc ctgggccaca gctgccagtg gaccgcagca ctctactccc gagacatgcg
     1741 ctttgtgcac taccagaccc tgagagccct attccagtac aaaccccaga acgtggatga
     1801 gctgacgcta agtcctggtg actacatctt tgtggacccc acgcagcagg acgaagccag
     1861 cgagggctgg gtgattggga tctcacagcg gacgggctgc cggggcttcc tgccggaaaa
     1921 ctacacggat cgagccagtg agtctgacac gtgggtgaag cacaggatgt acaccttcag
     1981 tctagccaca gacctgaact ccagaaagga tggtgaagcc agcagcagat gcagcgggga
     2041 atttcttcca caaacggcaa ggagtcttag cagcttacag gccttgcagg ctaccgttgc
     2101 aaggaagagt gtgctggtgg ttcgccacgg ggagagagtg gatcagatct tcgggaaggc
     2161 atggctgcag caatgctcca ctcctgatgg gaaatactac aggccagacc tgaatttccc
     2221 ctgcagtctg cccagacgga gtcgtgggat caaagacttt gaaaacgatc ccccattatc
     2281 atcgtgtggc attttccagt ccagaattgc aggggacgcg ctactggaca gtggtatcag
     2341 aatcagctct gtgtttgcct ccccagccct ccgctgtgtg cagacggcca aactcatcct
     2401 ggaagaactc aaactggaga aaaaaatcaa gatacgagtg gaacctggaa tctttgaatg
     2461 gacaaaatgg gaagctggca agaccacccc aaccctcatg agcctggaag agctgaaaga
     2521 ggcaaatttc aacattgaca ctgattacag gcccgcgttt cccctgtccg ccctcatgcc
     2581 ggccgagagc taccaggagt acatggacag gtgcacggcg agcatggtgc aaatcgtcaa
     2641 cacctgtccg caggacacgg gtgtcatcct aattgtgagt cacggctcca ctctggactc
     2701 ctgcacgcgg ccactgctcg ggctgccgcc ccgggaatgt ggggattttg cccaactcgt
     2761 gagaaagatc ccttccctgg gcatgtgctt ctgtgaagaa aataaagagg aaggaaaatg
     2821 ggagttggtg aacccaccgg tgaagaccct gacccacggg gcgaacgcag catttaactg
     2881 gaggaactgg atctcaggca actgaaaccc agctttcttg tacaaagtgg tgataaaccc
     2941 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3001 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3061 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3121 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     3181 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3241 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3301 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3361 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3421 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3481 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3541 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3601 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     3661 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3721 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3781 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3841 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3901 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     3961 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4021 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4081 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4141 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4201 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4261 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4321 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4381 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4441 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4501 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4561 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4621 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4681 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4741 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4801 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4861 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4921 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4981 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5041 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5101 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5161 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5221 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5281 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5341 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5401 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5461 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5521 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5581 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5641 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5701 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5761 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5821 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5881 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5941 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6001 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6061 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6121 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6181 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6241 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6301 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6361 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6421 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6481 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6541 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6601 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6661 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6721 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6781 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6841 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6901 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6961 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7021 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7081 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7141 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7201 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7261 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7321 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
