LOCUS       sequence-379883-        7473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human K202R 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-K202R-long-UBASH3A
FEATURES             Location/Qualifiers
     source          1..7473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        84..463
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        464..667
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     618..638
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     712..731
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        712..730
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             763..804
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             805..825
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    856..880
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2870..2894)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2913..2930)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2919..3143
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3189..3617
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3276..3295)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3486..3507
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3626..3646)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3631..3960
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3811..3946
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3873..3892
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4027..4821
                     /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(4081..4100)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4691..4710
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4995..5116
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5032..5051)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5086..5105
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5165..5181)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5165..5181)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5178..5200)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5189..5205
                     /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(5213..5219,5220..5237,5238..5243))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5258..5279
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5396..5413)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5567..6155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5647..5666)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6326..7117,7118..7186))
                     /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     6949..6968
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7187..7291)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7366..7385)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
       61 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
      121 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      181 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      241 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      301 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      361 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      421 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      481 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      541 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      601 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      661 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
      721 tcactatagg gagacccaag ctggctagtt aagcttacca tgggtaagcc tatccctaac
      781 cctctcctcg gtctcgattc tacggaaaac ctgtattttc agggcccgga tccgtcgacg
      841 aattctgcag atatcacaag tttgtacaaa aaagcaggct tcatggcagc gggggagacg
      901 cagctctacg ccaaggtctc caacaagctc aagagccgca gcagcccctc gctcctggag
      961 cccctcctgg ccatgggctt cccggtgcac accgcgctga aagcgttggc agccacgggg
     1021 aggaagacgg cggaggaggc cttggcctgg ctgcatgatc attgcaatga cccttcccta
     1081 gacgacccca tcccccagga gtatgccctt ttcctctgtc caacggggcc cctgctggaa
     1141 aaacttcaag agttctggag agagagcaag cgccagtgtg caaagaacag agctcatgag
     1201 gtcttcccac acgtgacact ctgtgacttc ttcacgtgtg aagaccagaa ggtggaatgc
     1261 ctgtacgagg cgctgaagag agctggagac aggctcctgg gctccttccc cacggccgtg
     1321 cctctggctc tccactcctc catcagctac ctcggcttct tcgtcagtgg cagccccgca
     1381 gacgtcatcc gggaattcgc catgaccttc gccacggaag catctctctt agcaggcact
     1441 tccgtttccc gcttctggat tttcagccag gtgcctggac atggccctaa cctgaggctg
     1501 agcaatttaa ctagagcctc cttcgtgagc cactacatcc ttcaaaaata ctgctccgtg
     1561 aagccttgca ccaaacagct gcatctgacc ttggcccaca ggttctaccc ccaccaccag
     1621 aggacgctgg agcagctggc cagagccatc cccctgggcc acagctgcca gtggaccgca
     1681 gcactctact cccgagacat gcgctttgtg cactaccaga ccctgagagc cctattccag
     1741 tacaaacccc agaacgtgga tgagctgacg ctaagtcctg gtgactacat ctttgtggac
     1801 cccacgcagc aggacgaagc cagcgagggc tgggtgattg ggatctcaca gcggacgggc
     1861 tgccggggct tcctgccgga aaactacacg gatcgagcca gtgagtctga cacgtgggtg
     1921 aagcacagga tgtacacctt cagtctagcc acagacctga actccagaaa ggatggtgaa
     1981 gccagcagca gatgcagcgg ggaatttctt ccacaaacgg caaggagtct tagcagctta
     2041 caggccttgc aggctaccgt tgcaaggaag agtgtgctgg tggttcgcca cggggagaga
     2101 gtggatcaga tcttcgggaa ggcatggctg cagcaatgct ccactcctga tgggaaatac
     2161 tacaggccag acctgaattt cccctgcagt ctgcccaggc ggagtcgtgg gatcaaagac
     2221 tttgaaaacg atcccccatt atcatcgtgt ggcattttcc agtccagaat tgcaggggac
     2281 gcgctactgg acagtggtat cagaatcagc tctgtgtttg cctccccagc cctccgctgt
     2341 gtgcagacgg ccaaactcat cctggaagaa ctcaaactgg agaaaaaaat caagatacga
     2401 gtggaacctg gaatctttga atggacaaaa tgggaagctg gcaaaaccac cccaaccctc
     2461 atgagcctgg aagagctgaa agaggcaaat ttcaacattg acactgatta caggcccgcg
     2521 tttcccctgt ccgccctcat gccggccgag agctaccagg agtacatgga caggtgcacg
     2581 gcgagcatgg tgcaaatcgt caacacctgc ccgcaggaca cgggtgtcat cctaattgtg
     2641 agtcacggct ccactctgga ctcctgcacg cggccactgc tcgggctgcc gccccgggaa
     2701 tgtggggatt ttgcccaact cgtgagaaag atcccttccc tgggcatgtg cttctgtgaa
     2761 gaaaataaag aggaaggaaa atgggagttg gtgaacccac cggtgaagac cctgacccac
     2821 ggggcgaacg cagcatttaa ctggaggaac tggatctcag gcaactgaaa cccagctttc
     2881 ttgtacaaag tggtgataaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     2941 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3001 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3061 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3121 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3181 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3241 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3301 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3361 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3421 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3481 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3541 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3601 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3661 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3721 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3781 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3841 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3901 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3961 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4021 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4081 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4141 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4201 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4261 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4321 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4381 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4441 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4501 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4561 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4621 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4681 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4741 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4801 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4861 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4921 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4981 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5041 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5101 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5161 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5221 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5281 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5341 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5401 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5461 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5521 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5581 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5641 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5701 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5761 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5821 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5881 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5941 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6001 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6061 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6121 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6181 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6241 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6301 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6361 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6421 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6481 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6541 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6601 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6661 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6721 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6781 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6841 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6901 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6961 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7021 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7081 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7141 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7201 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7261 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7321 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     7381 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     7441 cgcgagcaaa atttaagcta caacaaggca agg
//
