LOCUS       sequence-311793-        6322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of 6xHis HOXD13 (+7A) IDR mCherry Fusion 
            Protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6322)
  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 6322)
  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(+7A)-IDR-mCherry
FEATURES             Location/Qualifiers
     source          1..6322
                     /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             1230..1274
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1312..1330)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1326..1373
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1410..1865
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1497..1516)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1707..1728
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1980..2771,2772..2840))
                     /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     2603..2622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2841..2933)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3014..3602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3503..3522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3756..3773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3788..3927
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3853..3872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4013..4035
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4029..4220)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4792..5874)
                     /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     5836..5855
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5875..5952)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6158..6177
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6265..6284
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6265..6283
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6284..6308
                     /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 ccggtgacag tggatccgga gctaccatgt cccgagccgg tagctgggac atggacggcc
      121 tgcgagcgga cggaggcgga gctggaggag ctcccgcatc aagtagtagc tcaagtgtgg
      181 ccgctgccgc ggctagcgga caatgtaggg ggtttttgtc agcgcctgtc tttgcgggca
      241 cacattccgg gagggccgct gctgcggctg ctgctgctgc tgcagccgcg gcggcagctg
      301 cagctgcggc cgctgccgct agtggatttg cctatcctgg gacgagtgag cgcactggtt
      361 catcatcctc atcttcatcc agtgcggtag tcgccgctcg gccagaagca ccacctgcaa
      421 aagagtgtcc ggccccgacg ccagctgcag cagcagcggc accgccttca gctcctgcgg
      481 gtaccggagg cgggatggtt tccaaaggag aggaggataa tatggctata attaaagagt
      541 ttatgcggtt caaggtgcat atggagggat ctgtgaacgg tcacgagttc gaaatcgagg
      601 gcgaaggcga ggggcgaccc tatgaaggga cacagacggc caaactgaag gtgaccaagg
      661 gtggccccct gcctttcgcc tgggacatcc tgtcacccca gtttatgtac ggctctaagg
      721 cttacgttaa gcaccctgcc gatattccgg actacttgaa actgagcttc cctgaagggt
      781 tcaagtggga acgggtaatg aattttgagg atggcggtgt agtcacagtt acccaggaca
      841 gctctcttca ggacggagaa tttatctata aggttaaact tcggggcact aacttcccat
      901 cagacggccc cgtgatgcag aagaaaacta tgggctggga agccagctca gaacgcatgt
      961 atcccgagga cggagcactg aagggagaaa tcaagcagcg actcaagctg aaggatgggg
     1021 gacattatga tgctgaggtc aaaaccacct ataaggccaa gaaacccgtt cagcttcctg
     1081 gggcatacaa cgtgaatatc aaactggata ttacatccca taacgaagac tataccatcg
     1141 tggaacagta cgagcgggcc gagggcagac atagcacagg gggcatggat gaattgtaca
     1201 agaagcttgc ggccgcactc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc
     1261 agcacatgga ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc
     1321 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     1381 gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     1441 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1501 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1561 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1621 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1681 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1741 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     1801 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     1861 aatttctggc ggcacgatgg catgagatta tcaaaaagga tcttcaccta gatcctttta
     1921 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1981 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2041 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2101 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2161 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2221 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2281 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2341 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2401 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2461 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2521 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2581 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2641 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2701 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2761 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2821 cggaaatgtt gaatactcat actcttcctt tttcaatcat gattgaagca tttatcaggg
     2881 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggtca
     2941 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3001 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3061 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3121 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3721 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3781 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     3841 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     3901 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     3961 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4021 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     4081 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     4141 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     4201 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     4261 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     4321 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     4381 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     4441 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     4501 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     4561 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     4621 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcta
     4681 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     4741 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     4801 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4861 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     4921 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     4981 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     5041 gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc
     5101 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     5161 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     5221 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     5281 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     5341 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     5401 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     5461 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     5521 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     5581 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     5641 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     5701 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     5761 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     5821 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     5881 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     5941 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     6001 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     6061 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     6121 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     6181 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     6241 agatcgatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
     6301 acaattcccc tctagaaata at
//
