LOCUS       sequence-311789-        5899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of  6xHis mCherry Fusion Protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5899)
  AUTHORS   Basu S, Mackowiak SD, Niskanen H, Knezevic D, Asimi V, Grosswendt S,
            Geertsema H, Ali S, Jerkovic I, Ewers H, Mundlos S, Meissner A, 
            Ibrahim DM, Hnisz D
  TITLE     Unblending of Transcriptional Condensates in Human Repeat Expansion 
            Disease.
  JOURNAL   Cell. 2020 May 28;181(5):1062-1079.e30. doi: 
            10.1016/j.cell.2020.04.018. Epub 2020 May 7.
  PUBMED    32386547
REFERENCE   2  (bases 1 to 5899)
  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.cell.2020.04.018"; journalName: "Cell"; date: 
            "2020-05-28- 28"; volume: "181"; issue: "5"; pages: "1062-1079.e30"
COMMENT     Sequence Label: pET-45b-mCherry
FEATURES             Location/Qualifiers
     source          1..5899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             36..53
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             807..851
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(889..907)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      903..950
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      987..1442
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1074..1093)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1284..1305
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1557..2348,2349..2417))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2180..2199
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2418..2510)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2591..3179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3080..3099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3333..3350
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3365..3504
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3430..3449)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3590..3612
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3606..3797)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4369..5451)
                     /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     5413..5432
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5452..5529)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5735..5754
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5842..5861
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5842..5860
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5861..5885
                     /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)."
ORIGIN
        1 tttgtttaac tttaagaagg agatatacca tggcacatca ccaccaccat cacgtgggta
       61 ccggaggcgg gatggtttcc aaaggagagg aggataatat ggctataatt aaagagttta
      121 tgcggttcaa ggtgcatatg gagggatctg tgaacggtca cgagttcgaa atcgagggcg
      181 aaggcgaggg gcgaccctat gaagggacac agacggccaa actgaaggtg accaagggtg
      241 gccccctgcc tttcgcctgg gacatcctgt caccccagtt tatgtacggc tctaaggctt
      301 acgttaagca ccctgccgat attccggact acttgaaact gagcttccct gaagggttca
      361 agtgggaacg ggtaatgaat tttgaggatg gcggtgtagt cacagttacc caggacagct
      421 ctcttcagga cggagaattt atctataagg ttaaacttcg gggcactaac ttcccatcag
      481 acggccccgt gatgcagaag aaaactatgg gctgggaagc cagctcagaa cgcatgtatc
      541 ccgaggacgg agcactgaag ggagaaatca agcagcgact caagctgaag gatgggggac
      601 attatgatgc tgaggtcaaa accacctata aggccaagaa acccgttcag cttcctgggg
      661 catacaacgt gaatatcaaa ctggatatta catcccataa cgaagactat accatcgtgg
      721 aacagtacga gcgggccgag ggcagacata gcacaggggg catggatgaa ttgtacaaga
      781 agcttgcggc cgcactcgag tctggtaaag aaaccgctgc tgcgaaattt gaacgccagc
      841 acatggactc gtctactagc gcagcttaat taacctaggc tgctgccacc gctgagcaat
      901 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      961 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     1021 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1081 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1141 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1201 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1261 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1321 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1381 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     1441 ttctggcggc acgatggcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1501 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1561 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1621 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1681 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     1741 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1801 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1861 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     1921 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     1981 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2041 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     2101 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     2161 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     2221 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     2281 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     2341 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     2401 aaatgttgaa tactcatact cttccttttt caatcatgat tgaagcattt atcagggtta
     2461 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggtcatga
     2521 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2581 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2641 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2701 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     2761 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2821 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2881 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2941 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3001 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3061 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3121 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3181 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3241 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3301 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3361 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     3421 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     3481 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     3541 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3601 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     3661 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     3721 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     3781 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     3841 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     3901 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     3961 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     4021 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     4081 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     4141 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     4201 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgctagtc
     4261 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     4321 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     4381 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4441 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     4501 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     4561 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     4621 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
     4681 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     4741 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     4801 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     4861 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     4921 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     4981 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     5041 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     5101 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     5161 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     5221 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     5281 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     5341 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     5401 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     5461 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     5521 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     5581 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     5641 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     5701 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     5761 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     5821 tcgatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     5881 attcccctct agaaataat
//
