LOCUS       sequence-355927-        6129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein purification in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6129)
  AUTHORS   Wu M, Xu G, Han C, Luan PF, Xing YH, Nan F, Yang LZ, Huang Y, Yang 
            ZH, Shan L, Yang L, Liu J, Chen LL
  TITLE     lncRNA SLERT controls phase separation of FC/DFCs to facilitate Pol 
            I transcription.
  JOURNAL   Science. 2021 Jul 30;373(6554):547-555. doi: 
            10.1126/science.abf6582.
  PUBMED    34326237
REFERENCE   2  (bases 1 to 6129)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2021-07-30- 30"; volume: "373"; 
            issue: "6554"; pages: "547-555"
COMMENT     Sequence Label: PGEX-DDX21-D2+C
FEATURES             Location/Qualifiers
     source          1..6129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             942..965
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2179..2201)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2346..2364)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2432..2536
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2537..2605,2606..3397)
                     /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(2755..2774)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3568..4156
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4057..4076
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4310..4327
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4400..4477
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4478..5560
                     /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(4497..4516)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5609..5614,5615..5632,5633..5639)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5647..5663
                     /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     5671..5687
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5671..5687
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5683..6129,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5703..5720)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5703..5719)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5712..5734)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gttccgcgtg gatccccgat ggactacaaa gacgatgacg
      961 ataaggtgga cctgattggt aaaaagactc agaaaacggc aataactgtg gagcatctgg
     1021 ctattaagtg ccactggact cagagggcag cagttattgg ggatgtcatc cgagtatata
     1081 gtggtcatca aggacgcact atcatctttt gtgaaaccaa gaaagaagcc caggagctgt
     1141 cccagaattc agctataaag caggatgctc agtccttgca tggagacatt ccacagaagc
     1201 aaagggaaat caccctgaaa ggttttagaa atggtagttt tggagttttg gtggcaacca
     1261 atgttgctgc acgtgggtta gacatccctg aggttgattt ggttatacaa agctctccac
     1321 caaaggatgt agagtcctac attcatcgat ccgggcggac aggcagagct ggaaggacgg
     1381 gggtgtgcat ctgcttttat cagcacaagg aagaatatca gttagtacaa gtggagcaaa
     1441 aagcgggaat taagttcaaa cgaataggtg ttccttctgc aacagaaata ataaaagctt
     1501 ccagcaaaga tgccatcagg cttttggatt ccgtgcctcc cactgccatt agtcacttca
     1561 aacaatcagc tgagaagctg atagaggaga agggagctgt ggaagctctg gcagcagcac
     1621 tggcccatat ttcaggtgcc acgtccgtag accagcgctc cttgatcaac tcaaatgtgg
     1681 gttttgtgac catgatcttg cagtgctcaa ttgaaatgcc aaatattagt tatgcttgga
     1741 aagaacttaa agagcagctg ggcgaggaga ttgattccaa agtgaaggga atggtttttc
     1801 tcaaaggaaa gctgggtgtt tgctttgatg tacctaccgc atcagtaaca gaaatacagg
     1861 agaaatggca tgattcacga cgctggcagc tctctgtggc cacagagcaa ccagaactgg
     1921 aaggaccacg ggaaggatat ggaggcttca ggggacagcg ggaaggcagt cgaggcttca
     1981 ggggacagcg ggacggaaac agaagattca gaggacagcg ggaaggcagt agaggcccga
     2041 gaggacagcg atcaggaggt ggcaacaaaa gtaacagatc ccaaaacaaa ggccagaagc
     2101 ggagtttcag taaagcattt ggtcaaatcg tgactgactg acgatctgcc tcgcgcgttt
     2161 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     2221 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     2281 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg
     2341 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2401 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2461 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2521 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2581 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2641 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2701 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2761 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2821 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2881 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2941 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3001 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3061 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     3121 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3181 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3241 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3301 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3361 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3421 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3481 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3541 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3601 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3661 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     3721 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3781 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3841 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3901 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3961 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4021 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4081 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4141 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4201 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4261 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4321 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4381 ttcacaccgc ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
     4441 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     4501 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     4561 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     4621 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     4681 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     4741 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     4801 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     4861 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     4921 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     4981 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     5041 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     5101 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     5161 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     5221 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     5281 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     5341 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     5401 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     5461 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5521 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
     5581 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     5641 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat
     5701 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     5761 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     5821 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca
     5881 ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc
     5941 gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat
     6001 cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc
     6061 acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc
     6121 gttggaatt
//
