LOCUS       sequence-311802-        6526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of 6xHis HOXA13(WT) IDR mCherry Fusion Protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6526)
  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 6526)
  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-HOXA13(WT)-IDR-mCherry
FEATURES             Location/Qualifiers
     source          1..6526
                     /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             1434..1478
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1516..1534)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1530..1577
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1614..2069
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1701..1720)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1911..1932
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2184..2975,2976..3044))
                     /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     2807..2826
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3045..3137)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3218..3806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3707..3726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3960..3977
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3992..4131
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4057..4076)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4217..4239
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4233..4424)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4996..6078)
                     /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     6040..6059
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6079..6156)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6362..6381
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6469..6488
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6469..6487
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6488..6512
                     /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 gctaccatgg agttgaacaa aaacatggag ggggcggctg
      121 cggctgcagc cgcggcagcg gctgcagcgg ctgcaggagc cggtggagga ggttttcccc
      181 atccggcggc ggctgccgcg ggggggaatt tcagtgttgc ggcagctgca gcagctgcag
      241 ctgctgccgc agctaaccag tgccgcaacc tgatggcgca tccagcgcct cttgcgccgg
      301 gggccgcatc agcgtacagt tctgctcctg gggaagcacc cccgtccgcg gctgctgccg
      361 cggcagcggc agcagcagcc gcggctgcgg ccgcggcagc tagctccagt ggaggtcccg
      421 gaccagcggg acctgctggt gcggaagccg cgaagcagtg tagcccgtgc agcgcggcag
      481 ctcagagctc atccggtccc gccgccctcc catacggata ctttggctca gggtactacc
      541 cgtgcgcgag aatgggccct caccccaacg ccataaagtc atgtgcccaa cccgcgtcag
      601 ccgcagcggc agcagctttt gccgataagt acatggacac tgctggcccc gcggcggagg
      661 agttcagtag cagggcgaag gagggtaccg gaggcgggat ggtttccaaa ggagaggagg
      721 ataatatggc tataattaaa gagtttatgc ggttcaaggt gcatatggag ggatctgtga
      781 acggtcacga gttcgaaatc gagggcgaag gcgaggggcg accctatgaa gggacacaga
      841 cggccaaact gaaggtgacc aagggtggcc ccctgccttt cgcctgggac atcctgtcac
      901 cccagtttat gtacggctct aaggcttacg ttaagcaccc tgccgatatt ccggactact
      961 tgaaactgag cttccctgaa gggttcaagt gggaacgggt aatgaatttt gaggatggcg
     1021 gtgtagtcac agttacccag gacagctctc ttcaggacgg agaatttatc tataaggtta
     1081 aacttcgggg cactaacttc ccatcagacg gccccgtgat gcagaagaaa actatgggct
     1141 gggaagccag ctcagaacgc atgtatcccg aggacggagc actgaaggga gaaatcaagc
     1201 agcgactcaa gctgaaggat gggggacatt atgatgctga ggtcaaaacc acctataagg
     1261 ccaagaaacc cgttcagctt cctggggcat acaacgtgaa tatcaaactg gatattacat
     1321 cccataacga agactatacc atcgtggaac agtacgagcg ggccgagggc agacatagca
     1381 cagggggcat ggatgaattg tacaagaagc ttgcggccgc actcgagtct ggtaaagaaa
     1441 ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc tactagcgca gcttaattaa
     1501 cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1561 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc
     1621 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1681 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1741 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1801 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1861 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1921 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1981 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2041 ttttaacaaa atattaacgt ttacaatttc tggcggcacg atggcatgag attatcaaaa
     2101 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2161 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2221 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2281 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2341 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2401 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2461 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2521 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2581 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2641 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2701 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2761 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     2821 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2881 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     2941 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3001 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3061 tcatgattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3121 ttagaaaaat aaacaaatag gtcatgacca aaatccctta acgtgagttt tcgttccact
     3181 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3241 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3301 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3361 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3421 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3481 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3541 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3601 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3661 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3721 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3781 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3841 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3901 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3961 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     4021 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     4081 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4141 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4201 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4261 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     4321 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     4381 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     4441 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     4501 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     4561 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     4621 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     4681 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     4741 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     4801 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     4861 aacgacagga gcacgatcat gctagtcatg ccccgcgccc accggaagga gctgactggg
     4921 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     4981 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5041 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     5101 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     5161 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     5221 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatgtc
     5281 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     5341 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     5401 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     5461 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     5521 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     5581 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     5641 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     5701 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     5761 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     5821 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     5881 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     5941 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     6001 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     6061 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     6121 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     6181 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     6241 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     6301 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     6361 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     6421 cgatgcgtcc ggcgtagagg atcgagatcg atctcgatcc cgcgaaatta atacgactca
     6481 ctatagggga attgtgagcg gataacaatt cccctctaga aataat
//
