LOCUS       sequence-367254-        5137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of GST-tagged PML RING domain in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5137)
  AUTHORS   Bregnard TA, Fairchild D, Erlandsen H, Semenova IV, Szczepaniak R, 
            Ahmed A, Weller SK, Korzhnev DM, Bezsonova I
  TITLE     Conformational exchange at a C(2)H(2) zinc-binding site facilitates 
            redox sensing by the PML protein.
  JOURNAL   Structure. 2023 Sep 7;31(9):1086-1099.e6. doi: 
            10.1016/j.str.2023.06.014. Epub 2023 Jul 19.
  PUBMED    37473756
REFERENCE   2  (bases 1 to 5137)
  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"; doi: 
            "10.1016/j.str.2023.06.014"; journalName: "Structure"; date: 
            "2023-09-7- 7"; volume: "31"; issue: "9"; pages: "1086-1099.e6"
COMMENT     Sequence Label: pGEX-PML RING
FEATURES             Location/Qualifiers
     source          1..5137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..1083
                     /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(20..39)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1132..1137,1138..1155,1156..1162)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1170..1186
                     /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     1194..1210
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1194..1210
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1206..1733
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1226..1243)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1226..1242)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1235..1257)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1835..1840,1841..1856,1857..1863)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1871..1887
                     /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             1910..2563
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2521..2543
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2570..2593
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2839..2861)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3006..3024)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3092..3196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3197..3265,3266..4057)
                     /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(3415..3434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4228..4816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4717..4736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4970..4987
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5060..5137
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
       61 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
      121 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
      181 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
      241 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
      301 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
      361 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
      421 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
      481 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
      541 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
      601 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
      661 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
      721 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
      781 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
      841 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
      901 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
      961 gtctcactgg tgaaaagaaa aaccacgcta gcgcccaata cgcaaaccgc ctctccccgc
     1021 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     1081 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg ctttacactt
     1141 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     1201 cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     1261 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     1321 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     1381 ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg gagtgcgatc
     1441 ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt tacgatgcgc
     1501 ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt cccacggaga
     1561 atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta caggaaggcc
     1621 agacgcgaat tatttttgat ggcgttggaa ttagcttatc gactgcacgg tgcaccaatg
     1681 cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg taaatcactg
     1741 cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg cgccgacatc
     1801 ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc ggctcgtata
     1861 atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagtattca tgtcccctat
     1921 actaggttat tggaaaatta agggccttgt gcaacccact cgacttcttt tggaatatct
     1981 tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa
     2041 aaagtttgaa ttgggtttgg agtttcccaa tcttccttat tatattgatg gtgatgttaa
     2101 attaacacag tctatggcca tcatacgtta tatagctgac aagcacaaca tgttgggtgg
     2161 ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga gcggttttgg atattagata
     2221 cggtgtttcg agaattgcat atagtaaaga ctttgaaact ctcaaagttg attttcttag
     2281 caagctacct gaaatgctga aaatgttcga agatcgttta tgtcataaaa catatttaaa
     2341 tggtgatcat gtaacccatc ctgacttcat gttgtatgac gctcttgatg ttgttttata
     2401 catggaccca atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa aacgtattga
     2461 agctatccca caaattgata agtacttgaa atccagcaag tatatagcat ggcctttgca
     2521 gggctggcaa gccacgtttg gtggtggcga ccatcctcca aaatcggatc tggaagttct
     2581 gttccagggg cccctgggat ccgaggaaga gttccagttt ctgcgttgcc agcaatgcca
     2641 agcggaagcg aagtgcccga aactgctgcc gtgcctgcac accctgtgca gcggttgcct
     2701 ggaggcgagc ggtatgcagt gcccgatctg ccaagcgccg tggccgctgg gtgcggacac
     2761 cccggcgctg taactcgagc ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt
     2821 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     2881 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     2941 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg
     3001 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3061 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3121 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3181 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3241 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3301 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3361 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3421 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3481 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3541 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3601 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3661 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3721 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     3781 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3841 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3901 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3961 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4021 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4081 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4141 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4201 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4261 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4321 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4381 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4441 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4501 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4561 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4621 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4681 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4741 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4801 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4861 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4921 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4981 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     5041 ttcacaccgc ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
     5101 atagcgcccg gaagagagtc aattcagggt ggtgaat
//
