LOCUS       sequence-379911-        6838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human wild-type short-form UBASH3A with GST 
            tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6838)
  TITLE     Concannon Lab UBASH3A Plasmids 2022
REFERENCE   2  (bases 1 to 6838)
  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: pGEX-6P-1-WT-short-UBASH3A
FEATURES             Location/Qualifiers
     source          1..6838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(540..545,546..563,564..570)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    578..594
                     /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)."
     primer_bind     602..618
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     602..618
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             614..1141
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(634..651)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(634..650)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(643..665)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1243..1248,1249..1264,1265..1271)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1279..1295
                     /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)."
     CDS             1318..1971
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1929..1951
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1978..2001
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3948..3970)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4115..4133)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4201..4305
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4306..4374,4375..5166)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4524..4543)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5337..5925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5826..5845
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6079..6096
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6169..6246
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(6247..6838,1..491)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(6266..6285)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
       61 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
      121 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
      181 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
      241 acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
      301 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
      361 tgttgcccgt ctcactggtg aaaagaaaaa ccacgctagc gcccaatacg caaaccgcct
      421 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
      481 gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc accccaggct
      541 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
      601 acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac aacgtcgtga
      661 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
      721 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
      781 tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga
      841 gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta
      901 cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc
      961 cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca
     1021 ggaaggccag acgcgaatta tttttgatgg cgttggaatt agcttatcga ctgcacggtg
     1081 caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta
     1141 aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg
     1201 ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatcgg
     1261 ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac agtattcatg
     1321 tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg
     1381 gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg
     1441 cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt
     1501 gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg
     1561 ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat
     1621 attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat
     1681 tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca
     1741 tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt
     1801 gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa
     1861 cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg
     1921 cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatctg
     1981 gaagttctgt tccaggggcc cctgggatcc atggcagcgg gggagacgca gctctacgcc
     2041 aaggtctcca acaagctcaa gagccgcagc agcccctcgc tcctggagcc cctcctggcc
     2101 atgggcttcc cggtgcacac cgcgctgaaa gcgttggcag ccacggggag gaagacggcg
     2161 gaggaggcct tggcctggct gcatgatcat tgcaatgacc cttccctaga cgaccccatc
     2221 ccccaggagt atgccctttt cctctgtcca acggggcccc tgctggaaaa acttcaagag
     2281 ttctggagag agagcaagcg ccagtgtgca aagaacagag ctcatgaggt cttcccacac
     2341 gtgacactct gtgacttctt cacgtgtgaa gaccagaagg tggaatgcct gtacgaggcg
     2401 ctgaagagag ctggagacag gctcctgggc tccttcccca cggccgtgcc tctggctctc
     2461 cactcctcca tcagctacct cggcttcttc gtcagtggca gccccgcaga cgtcatccgg
     2521 gaattcgcca tgaccttcgc cacggaagca tctctcttag cagactgctc cgtgaagcct
     2581 tgcaccaaac agctgcatct gaccttggcc cacaagttct acccccacca ccagaggacg
     2641 ctggagcagc tggccagagc catccccctg ggccacagct gccagtggac cgcagcactc
     2701 tactcccgag acatgcgctt tgtgcactac cagaccctga gagccctatt ccagtacaaa
     2761 ccccagaacg tggatgagct gacgctaagt cctggtgact acatctttgt ggaccccacg
     2821 cagcaggacg aagccagcga gggctgggtg attgggatct cacagcggac gggctgccgg
     2881 ggcttcctgc cggaaaacta cacggatcga gccagtgagt ctgacacgtg ggtgaagcac
     2941 aggatgtaca ccttcagtct agccacagac ctgaactcca gaaaggatgg tgaagccagc
     3001 agcagatgca gcggggaatt tcttccacaa acggcaagga gtcttagcag cttacaggcc
     3061 ttgcaggcta ccgttgcaag gaagagtgtg ctggtggttc gccacgggga gagagtggat
     3121 cagatcttcg ggaaggcatg gctgcagcaa tgctccactc ctgatgggaa atactacagg
     3181 ccagacctga atttcccctg cagtctgccc agacggagtc gtgggatcaa agactttgaa
     3241 aacgatcccc cattatcatc gtgtggcatt ttccagtcca gaattgcagg ggacgcgcta
     3301 ctggacagtg gtatcagaat cagctctgtg tttgcctccc cagccctccg ctgtgtgcag
     3361 acggccaaac tcatcctgga agaactcaaa ctggagaaaa aaatcaagat acgagtggaa
     3421 cctggaatct ttgaatggac aaaatgggaa gctggcaaga ccaccccaac cctcatgagc
     3481 ctggaagagc tgaaagaggc aaatttcaac attgacactg attacaggcc cgcgtttccc
     3541 ctgtccgccc tcatgccggc cgagagctac caggagtaca tggacaggtg cacggcgagc
     3601 atggtgcaaa tcgtcaacac ctgtccgcag gacacgggtg tcatcctaat tgtgagtcac
     3661 ggctccactc tggactcctg cacgcggcca ctgctcgggc tgccgccccg ggaatgtggg
     3721 gattttgccc aactcgtgag aaagatccct tccctgggca tgtgcttctg tgaagaaaat
     3781 aaagaggaag gaaaatggga gttggtgaac ccaccggtga agaccctgac ccacggggcg
     3841 aacgcagcat ttaactggag gaactggatc tcaggcaact gactcgagcg gccgcatcgt
     3901 gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     3961 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4021 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     4081 gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct atttttatag
     4141 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4201 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4261 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4321 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4381 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4441 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4501 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     4561 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4621 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4681 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4741 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4801 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     4861 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4921 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4981 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5041 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5101 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5161 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5221 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5281 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5341 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5401 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5461 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5521 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5581 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5641 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5701 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5761 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5821 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5881 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5941 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6001 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6061 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     6121 tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga caccatcgaa
     6181 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     6241 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     6301 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     6361 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     6421 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     6481 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     6541 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     6601 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     6661 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     6721 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     6781 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtc
//
