LOCUS       sequence-318747-        6301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of  6xHis HOXD13 IDR mCherry Fusion Protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6301)
  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 6301)
  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-HOXD13-IDR-mCherry
FEATURES             Location/Qualifiers
     source          1..6301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             94..111
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1267..1311
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1349..1367)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1363..1410
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1447..1902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1534..1553)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1744..1765
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2017..2808,2809..2877))
                     /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     2640..2659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2878..2970)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3051..3639
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3540..3559
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3793..3810
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3825..3964
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3890..3909)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4050..4072
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4066..4257)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4829..5911)
                     /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     5873..5892
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5912..5989)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6195..6214
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg gcacatcacc accaccatca cgtgggtacc
      121 ggtgacagtg gatccggagc taccatgtcc cgagccggta gctgggacat ggacggcctg
      181 cgagcggacg gaggcggagc tggaggagct cccgcatcaa gtagtagctc aagtgtggcc
      241 gctgccgcgg ctagcggaca atgtaggggg tttttgtcag cgcctgtctt tgcgggcaca
      301 cattccggga gggccgctgc tgcggctgct gctgctgctg cagccgcggc ggccgctagt
      361 ggatttgcct atcctgggac gagtgagcgc actggttcat catcctcatc ttcatccagt
      421 gcggtagtcg ccgctcggcc agaagcacca cctgcaaaag agtgtccggc cccgacgcca
      481 gctgcagcag cagcggcacc gccttcagct cctgcgggta ccggaggcgg gatggtttcc
      541 aaaggagagg aggataatat ggctataatt aaagagttta tgcggttcaa ggtgcatatg
      601 gagggatctg tgaacggtca cgagttcgaa atcgagggcg aaggcgaggg gcgaccctat
      661 gaagggacac agacggccaa actgaaggtg accaagggtg gccccctgcc tttcgcctgg
      721 gacatcctgt caccccagtt tatgtacggc tctaaggctt acgttaagca ccctgccgat
      781 attccggact acttgaaact gagcttccct gaagggttca agtgggaacg ggtaatgaat
      841 tttgaggatg gcggtgtagt cacagttacc caggacagct ctcttcagga cggagaattt
      901 atctataagg ttaaacttcg gggcactaac ttcccatcag acggccccgt gatgcagaag
      961 aaaactatgg gctgggaagc cagctcagaa cgcatgtatc ccgaggacgg agcactgaag
     1021 ggagaaatca agcagcgact caagctgaag gatgggggac attatgatgc tgaggtcaaa
     1081 accacctata aggccaagaa acccgttcag cttcctgggg catacaacgt gaatatcaaa
     1141 ctggatatta catcccataa cgaagactat accatcgtgg aacagtacga gcgggccgag
     1201 ggcagacata gcacaggggg catggatgaa ttgtacaaga agcttgcggc cgcactcgag
     1261 tctggtaaag aaaccgctgc tgcgaaattt gaacgccagc acatggactc gtctactagc
     1321 gcagcttaat taacctaggc tgctgccacc gctgagcaat aactagcata accccttggg
     1381 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg
     1441 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     1501 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     1561 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     1621 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     1681 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     1741 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     1801 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     1861 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttctggcggc acgatggcat
     1921 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1981 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2041 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2101 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2161 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2221 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2281 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2341 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2401 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2461 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2521 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2581 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2641 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2701 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2761 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2821 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2881 cttccttttt caatcatgat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2941 tatttgaatg tatttagaaa aataaacaaa taggtcatga ccaaaatccc ttaacgtgag
     3001 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3061 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3121 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3181 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3241 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3301 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3361 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3421 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3481 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3541 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3601 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3661 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3721 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3781 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3841 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
     3901 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     3961 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4021 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4081 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     4141 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     4201 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     4261 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     4321 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     4381 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     4441 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     4501 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     4561 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     4621 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     4681 tagccgggtc ctcaacgaca ggagcacgat catgctagtc atgccccgcg cccaccggaa
     4741 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     4801 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4861 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4921 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     4981 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     5041 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     5101 ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     5161 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     5221 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     5281 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     5341 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     5401 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     5461 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     5521 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     5581 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     5641 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     5701 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     5761 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     5821 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     5881 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     5941 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     6001 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     6061 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     6121 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     6181 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     6241 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tcgatctcga tcccgcgaaa
     6301 t
//
