LOCUS       sequence-326697-        8141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of a GST fusion of Human Uba1 with a 6alanine 
            linker between the GST encoded by the plasmid and the inserted Uba1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8141)
  TITLE     Haas Lab plasmids
REFERENCE   2  (bases 1 to 8141)
  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: pGEX4T-1-6AlanineHE1
FEATURES             Location/Qualifiers
     source          1..8141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     109..126
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        199..276
                     /gene="lacI"
                     /label=lacI promoter
     CDS             277..1359
                     /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(296..315)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1408..1413,1414..1431,1432..1438)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1446..1462
                     /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     1470..1486
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1470..1486
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1482..2009
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1502..1519)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1502..1518)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1511..1533)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2111..2116,2117..2132,2133..2139)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2147..2163
                     /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             2186..2839
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2797..2819
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2846..2863
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(6119..6141)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6286..6304)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6372..6476
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6477..6545,6546..7337)
                     /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(6695..6714)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7508..8096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7997..8016
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
       61 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
      121 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
      181 tcacaccgca taaattccga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga
      241 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat
      301 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
      361 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
      421 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
      481 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
      541 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
      601 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
      661 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
      721 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga cggtacgcga
      781 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
      841 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
      901 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
      961 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
     1021 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
     1081 tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc
     1141 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
     1201 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc
     1261 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     1321 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag
     1381 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     1441 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacggatt
     1501 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     1561 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     1621 gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg tttccggcac
     1681 cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat actgtcgtcg
     1741 tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac gtgacctatc
     1801 ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt tactcgctca
     1861 catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt tttgatggcg
     1921 ttggaattag cttatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg
     1981 aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc
     2041 actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg
     2101 aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa
     2161 caatttcaca caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg
     2221 ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt
     2281 gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt
     2341 tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat
     2401 acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat
     2461 ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag
     2521 taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat
     2581 gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga
     2641 cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc
     2701 gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta
     2761 cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg
     2821 tggcgaccat cctccaaaat cggatctggt tccgcgtgga tccgctgcag ctgcagcagc
     2881 tatgtccagc tcgccgctgt ccaagaaacg tcgcgtgtcc gggcctgatc caaagccggg
     2941 ttctaactgc tcccctgccc agtccgtgtt gtccgaagtg ccctcggtgc caaccaacgg
     3001 aatggccaag aacggcagtg aagcagacat agacgagggc ctttactccc ggcagctgta
     3061 tgtgttgggc catgaggcaa tgaagcggct ccagacatcc agtgtcctgg tatcaggcct
     3121 gcggggcctg ggcgtggaga tcgctaagaa catcatcctt ggtggggtca aggctgttac
     3181 cctacatgac cagggcactg cccagtgggc tgatctttcc tcccagttct acctgcggga
     3241 ggaggacatc ggtaaaaacc gggccgaggt atcacagccc cgcctcgctg agctcaacag
     3301 ctatgtgcct gtcactgcct acactggacc cctcgttgag gacttcctta gtggtttcca
     3361 ggtggtggtg ctcaccaaca cccccctgga ggaccagctg cgagtgggtg agttctgtca
     3421 caaccgtggc atcaagctgg tggtggcaga cacgcggggc ctgtttgggc agctcttctg
     3481 tgactttgga gaggaaatga tcctcacaga ttccaatggg gagcagccac tcagtgctat
     3541 ggtttctatg gttaccaagg acaaccccgg tgtggttacc tgcctggatg aggcccgaca
     3601 cgggtttgag agcggggact ttgtctcctt ttcagaagta cagggcatgg ttgaactcaa
     3661 cggaaatcag cccatggaga tcaaagtcct gggtccttat acctttagca tctgtgacac
     3721 ctccaacttc tccgactaca tccgtggagg catcgtcagt caggtcaaag tacctaagaa
     3781 gattagcttt aaatccttgg tggcctcact ggcagaacct gactttgtgg tgacggactt
     3841 cgccaagttt tctcgccctg cccagctgca cattggcttc caggccctgc accagttctg
     3901 tgctcagcat ggccggccac ctcggccccg caatgaggag gatgcagcag aactggtagc
     3961 cttagcacag gctgtgaatg ctcgagccct gccagcagtg cagcaaaata acctggacga
     4021 ggacctcatc cggaagctgg catatgtggc tgctggggat ctggcaccca taaacgcctt
     4081 cattgggggc ctggctgccc aggaagtcat gaaggcctgc tccgggaagt tcatgcccat
     4141 catgcagtgg ctatactttg atgcccttga gtgtctccct gaggacaaag aggtcctcac
     4201 agaggacaag tgcctccagc gccagaaccg ttatgacggg caagtggctg tgtttggctc
     4261 agacctgcaa gagaagctgg gcaagcagaa gtatttcctg gtgggtgcgg gggccattgg
     4321 ctgtgagctg ctcaagaact ttgccatgat tgggctgggc tgcggggagg gtggagaaat
     4381 catcgttaca gacatggaca ccattgagaa gtcaaatctg aatcgacagt ttcttttccg
     4441 gccctgggat gtcacgaagt taaagtctga cacggctgct gcagctgtgc gccaaatgaa
     4501 tccacatatc cgggtgacaa gccaccagaa ccgtgtgggt cctgacacgg agcgcatcta
     4561 tgatgacgat tttttccaaa acctagatgg cgtggccaat gccctggaca acgtggatgc
     4621 ccgcatgtac atggaccgcc gctgtgtcta ctaccggaag ccactgctgg agtcaggcac
     4681 actgggcacc aaaggcaatg tgcaggtggt gatccccttc ctgacagagt cgtacagttc
     4741 cagccaggac ccacctgaga agtccatccc catctgtacc ctgaagaact tccctaatgc
     4801 catcgagcac accctgcagt gggctcggga tgagtttgaa ggcctcttca agcagccagc
     4861 agaaaatgtc aaccagtacc tcacagaccc caagtttgtg gagcgaacac tgcggctggc
     4921 aggcactcag cccttggagg tgctggaggc tgtgcagcgc agcctggtgc tgcagcgacc
     4981 acagacctgg gctgactgcg tgacctgggc ctgccaccac tggcacaccc agtactcgaa
     5041 caacatccgg cagctgctgc acaacttccc tcctgaccag ctcacaagct caggagcgcc
     5101 gttctggtct gggcccaaac gctgtccaca cccgctcacc tttgatgtca acaatcccct
     5161 gcatctggac tatgtgatgg ctgctgccaa cctgtttgcc cagacctacg ggctgacagg
     5221 ctctcaggac cgagctgctg tggccacatt cctgcagtct gtgcaggtcc ccgaattcac
     5281 ccccaagtct ggcgtcaaga tccatgtttc tgaccaggag ctgcagagcg ccaatgcctc
     5341 tgttgatgac agtcgtctag aggagctcaa agccactctg cccagcccag acaagctccc
     5401 tggattcaag atgtacccca ttgactttga gaaggatgat gacagcaact ttcatatgga
     5461 tttcatcgtg gctgcatcca acctccgggc agaaaactat gacattcctt ctgcagaccg
     5521 gcacaagagc aagctgattg cagggaagat catcccagcc attgccacga ccacagcagc
     5581 cgtggttggc cttgtgtgtc tggaactgta caaggttgtg caggggcacc gacagcttga
     5641 ctcctacaag aatggtttcc tcaacttggc cctgcctttc tttggtttct ctgaacccct
     5701 tgccgcacca cgtcaccagt actataacca agagtggaca ttgtgggatc gctttgaggt
     5761 acaagggctg cagcctaatg gtgaggagat gaccctcaaa cagttcctcg actattttaa
     5821 gacagagcac aaattagaga tcaccatgct gtcccagggc gtgtccatgc tctattcctt
     5881 cttcatgcca gctgccaagc tcaaggaacg gttggatcag ccgatgacag agattgtgag
     5941 ccgtgtgtcg aagcgaaagc tgggccgcca cgtgcgggcg ctggtgcttg agctgtgctg
     6001 taacgacgag agcggcgagg atgtcgaggt tccctatgtc cgatacacca tccgctgagc
     6061 ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     6121 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     6181 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     6241 agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc
     6301 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6361 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6421 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6481 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6541 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6601 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6661 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     6721 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6781 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6841 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6901 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6961 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     7021 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7081 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7141 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     7201 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7261 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7321 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7381 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7441 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7501 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7561 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7621 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     7681 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7741 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7801 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7861 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7921 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7981 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8041 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8101 ccagcaacgc ggccttttta cggttcctgg ccttttgctg g
//
