LOCUS       sequence-367327-        5329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of GST-tagged PML fragment 49-168 (RING + B-box1) in 
            bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5329)
  AUTHORS   Bregnard T, Ahmed A, Semenova IV, Weller SK, Bezsonova I
  TITLE     The B-box1 domain of PML mediates SUMO E2-E3 complex formation 
            through an atypical interaction with UBC9.
  JOURNAL   Biophys Chem. 2022 Aug;287:106827. doi: 10.1016/j.bpc.2022.106827. 
            Epub 2022 May 18.
  PUBMED    35667129
REFERENCE   2  (bases 1 to 5329)
  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"; journalName: "Biophys 
            Chem."; date: "2022-08"; volume: "287:106827. doi"; pages: " 
            10.1016/j.bpc.2022.106827. Epub 2022 May 18"
COMMENT     Sequence Label: pGEX-PML RB1
FEATURES             Location/Qualifiers
     source          1..5329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..78
                     /gene="lacI"
                     /label=lacI promoter
     CDS             79..1161
                     /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(98..117)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1210..1215,1216..1233,1234..1240)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1248..1264
                     /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     1272..1288
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1272..1288
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1284..1811
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1304..1321)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1304..1320)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1313..1335)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1913..1918,1919..1934,1935..1941)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1949..1965
                     /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             1988..2641
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2599..2621
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2648..2671
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3109..3131)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3276..3294)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3362..3466
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3467..3535,3536..4327)
                     /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(3685..3704)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4498..5086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4987..5006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5240..5257
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
       61 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
      121 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
      181 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
      241 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
      301 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
      361 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
      421 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
      481 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
      541 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
      601 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
      661 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
      721 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
      781 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      841 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      901 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      961 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     1021 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccacgctagc gcccaatacg
     1081 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1141 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
     1201 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     1261 acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac
     1321 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     1381 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     1441 gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa
     1501 gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga
     1561 tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc
     1621 cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa
     1681 gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttggaatt agcttatcga
     1741 ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt
     1801 gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat
     1861 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat
     1921 taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     1981 agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg
     2041 acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg
     2101 tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta
     2161 tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa
     2221 gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc
     2281 ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct
     2341 caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg
     2401 tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc
     2461 tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg
     2521 ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta
     2581 tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa
     2641 atcggatctg gaagttctgt tccaggggcc cctgggatcc gaggaagagt tccagtttct
     2701 gcgttgccag caatgccaag cggaagcgaa gtgcccgaaa ctgctgccgt gcctgcacac
     2761 cctgtgcagc ggttgcctgg aggcgagcgg tatgcagtgc ccgatctgcc aagcgccgtg
     2821 gccgctgggt gcggacaccc cggcgctgga taacgttttc tttgaaagcc tgcagcgtcg
     2881 tctgagcgtg taccgtcaga ttgttgacgc gcaagcggtg tgcacccgtt gcaaggaaag
     2941 cgcggatttc tggtgctttg aatgcgagca gctgctgtgc gcgaagtgct tcgaggcgca
     3001 ccaatggttt ctgaagcatg aagcgcgtcc gctggcggag taactcgagc ggccgcatcg
     3061 tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     3121 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     3181 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     3241 cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     3301 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3361 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3421 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3481 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3541 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3601 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3661 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     3721 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3781 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3841 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3901 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3961 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     4021 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4081 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4141 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4201 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4261 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4321 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4381 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4441 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4501 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4561 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4621 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     4681 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4741 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4801 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4861 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4921 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4981 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5041 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5101 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5161 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5221 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     5281 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattcc
//
