LOCUS       sequence-355928-        5907 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 5907)
  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 5907)
  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: GST-FBL
FEATURES             Location/Qualifiers
     source          1..5907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(531..536,537..554,555..561)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    569..585
                     /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     593..609
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     593..609
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             605..1132
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(625..642)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(625..641)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(634..656)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1234..1239,1240..1255,1256..1262)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1270..1286
                     /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             1309..1962
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1920..1942
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1969..1986
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(2764..2775)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     primer_bind     complement(3008..3030)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3175..3193)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3261..3365
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3366..3434,3435..4226)
                     /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(3584..3603)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4397..4985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4886..4905
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5139..5156
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5229..5306
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5307..5907,1..482)
                     /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(5326..5345)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
       61 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
      121 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
      181 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat
      241 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
      301 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
      361 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
      421 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      481 gagcgcaacg caattaatgt aagttagctc actcattagg caccccaggc tttacacttt
      541 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
      601 agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg actgggaaaa
      661 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
      721 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
      781 gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg agtgcgatct
      841 tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt acgatgcgcc
      901 catctacacc aacgtgacct atcccattac ggtcaatccg ccgtttgttc ccacggagaa
      961 tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac aggaaggcca
     1021 gacgcgaatt atttttgatg gcgttggaat tagcttatcg actgcacggt gcaccaatgc
     1081 ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt aaatcactgc
     1141 ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc gccgacatca
     1201 taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg gctcgtataa
     1261 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagtattcat gtcccctata
     1321 ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt ggaatatctt
     1381 gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg gcgaaacaaa
     1441 aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg tgatgttaaa
     1501 ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat gttgggtggt
     1561 tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga tattagatac
     1621 ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga ttttcttagc
     1681 aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac atatttaaat
     1741 ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt tgttttatac
     1801 atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa acgtattgaa
     1861 gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg gcctttgcag
     1921 ggctggcaag ccacgtttgg tggtggcgac catcctccaa aatcggatct ggttccgcgt
     1981 ggatccccga tgaagccagg attcagtccc cgtgggggtg gctttggcgg ccgagggggc
     2041 tttggtgacc gtggtggtcg tggaggccga gggggctttg gcgggggccg aggtcgaggc
     2101 ggaggcttta gaggtcgtgg acgaggagga ggtggaggcg gcggcggcgg tggaggagga
     2161 ggaagaggtg gtggaggctt ccattctggt ggcaaccggg gtcgtggtcg gggaggaaaa
     2221 agaggaaacc agtcggggaa gaatgtgatg gtggagccgc atcggcatga gggtgtcttc
     2281 atttgtcgag gaaaggaaga tgcactggtc accaagaacc tggtccctgg ggaatcagtt
     2341 tatggagaga agagagtctc gatttcggaa ggagatgaca aaattgagta ccgagcctgg
     2401 aaccccttcc gctccaagct agcagcagca atcctgggtg gtgtggacca gatccacatc
     2461 aaaccggggg ctaaggttct ctacctcggg gctgcctcgg gcaccacggt ctcccatgtc
     2521 tctgacatcg ttggtccgga tggtctagtc tatgcagtcg agttctccca ccgctctggc
     2581 cgtgacctca ttaacttggc caagaagagg accaacatca ttcctgtgat cgaggatgct
     2641 cgacacccac acaaataccg catgctcatc gcaatggtgg atgtgatctt tgctgatgtg
     2701 gcccagccag accagacccg gattgtggcc ctgaatgccc acaccttcct gcgtaatgga
     2761 ggacactttg tgatttccat taaggccaac tgcattgact ccacagcctc agccgaggcc
     2821 gtgtttgcct ccgaagtgaa aaagatgcaa caggagaaca tgaagccgca ggagcagttg
     2881 acccttgagc catatgaaag agaccatgcc gtggtcgtgg gagtgtacag gccacccccc
     2941 aaggtgaaga actgaatcgt gactgactga cgatctgcct cgcgcgtttc ggtgatgacg
     3001 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     3061 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     3121 ccatgaccca gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg
     3181 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3241 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3301 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3361 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3421 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3481 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3541 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3601 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3661 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3721 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3781 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3841 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3901 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3961 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4021 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4081 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4141 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4201 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4261 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4321 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4381 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4441 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4501 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     4561 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4621 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4681 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4741 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4801 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4861 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4921 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4981 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5041 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5101 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5161 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5221 taaattccga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg
     5281 aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt
     5341 atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg
     5401 cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg
     5461 tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg
     5521 ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg
     5581 ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc
     5641 acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg
     5701 atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg
     5761 accagacacc catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg
     5821 agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg
     5881 tctcggcgcg tctgcgtctg gctggct
//
