LOCUS       sequence-471707-        7118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of mEGFP-NUP98-HOXA9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7118)
  AUTHORS   Zhang Y, Stoppelkamp I, Fernandez-Pernas P, Allram M, Charman M, 
            Magalhaes AP, Piedavent-Salomon M, Sommer G, Sung YC, Meyer K, Grams
            N, Halko E, Dongre S, Meierhofer D, Malszycki M, Ilik IA, Aktas T, 
            Kraushar ML, Vastenhouw N, Weitzman MD, Grebien F, Niskanen H, Hnisz
            D
  TITLE     Probing condensate microenvironments with a micropeptide killswitch.
  JOURNAL   Nature. 2025 Jun 4. doi: 10.1038/s41586-025-09141-5.
  PUBMED    40468084
REFERENCE   2  (bases 1 to 7118)
  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"; journalName: "Nature. 
            2025 Jun 4. doi: 10.1038/s41586-025-09141-5."
COMMENT     Sequence Label: pRK5_mEGFP-NUP98-HOXA9
FEATURES             Location/Qualifiers
     source          1..7118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      934..943
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(940..942,943..945,946..1656)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(985..1006)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1246..1265)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1593..1614
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      3418..3427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3431..3565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3468..3487)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3522..3541
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3634..3991
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3634..3825
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3657..3677)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3842..3977
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4011..4028)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4011..4027)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4020..4042)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4240..4695
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4327..4346)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4537..4558
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4712..4731)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4831..4853
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4891..4909)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4977..5081
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5082..5150,5151..5942)
                     /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     complement(5300..5319)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6113..6701
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6602..6621
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6855..6872
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6989..7010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7025..7030,7031..7048,7049..7055)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7063..7079
                     /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     7068..7090
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7087..7103
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7087..7103
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcgtaccg gtcgccacca tggtgagcaa gggcgaggag
      961 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1021 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1081 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1141 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1201 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1261 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1321 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1381 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1441 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1501 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
     1561 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1621 gccgggatca ctctcggcat ggacgagctg tacaagaccg gcggcatgtt taacaaatca
     1681 tttggaacac cctttggggg tggcacaggt ggctttggca caacttcaac atttggacag
     1741 aatactggct ttggcactac tagtggaggg gcatttggaa catctgcatt tggttctagc
     1801 aacaatactg gaggcctctt tggaaattca cagactaaac caggaggatt gtttggaacc
     1861 agttcattta gccagccagc tacctccaca agcactggct ttgggtttgg tacgtcaaca
     1921 ggaacagcaa ataccttgtt tggaactgca agcacaggga ccagtctctt ctcatcccaa
     1981 aacaatgcct ttgcacaaaa taaaccaact ggctttggca attttggaac cagtactagc
     2041 agtggaggac tctttggaac cacaaatacc acctctaatc cttttggcag cacatctggc
     2101 tccctctttg ggccaagtag ttttacagct gctcctactg ggactactat taaatttaac
     2161 cctccaactg gtacagatac tatggtcaaa gctggagtta gcactaacat aagtaccaag
     2221 caccagtgta ttactgctat gaaagaatat gaaagcaagt cactagagga acttcgttta
     2281 gaggattatc aggctaacag gaagggccca cagaaccagg tgggagcagg taccacaact
     2341 ggcttgtttg ggtcttctcc agccacttcc agcgcaacag gactcttcag ctcctccacc
     2401 actaattcag gctttgcata tggtcagaac aaaactgcct ttggaactag tacaactgga
     2461 tttggaacaa atccaggtgg tctctttggc caacagaatc agcagactac cagcctcttc
     2521 agcaaaccat ttggccaggc tacaaccacc cagaacactg gcttttcctt tggtaatacc
     2581 agcaccatag gacagccaag caccaacacc atgggattat ttggagtaac ccaagcctca
     2641 cagcctggag gtctttttgg gacagctaca aacaccagca ctgggacagc atttggaaca
     2701 ggaacaggtc tctttgggca gaccaatact ggatttggtg ctgttggttc gaccctgttt
     2761 ggcaataaca agcttactac atttggaagc agcacaacca gtgcaccttc atttggtaca
     2821 accagtggcg ggctctttgg ttttggcaca aataccagtg ggaatagtat ttttggaagt
     2881 aaaccagcac ctgggactct tggaactggg cttggtgcag gatttggaac agctcttggt
     2941 gctggacagg catctttgtt tgggaacaac caacctaaga ttggagggcc tcttggtaca
     3001 ggagcctttg gggcccctgg atttaatact acgacagcca ctttgggctt tggagccccc
     3061 caggccccag tagttgatag agaaaaacaa cccagcgaag gcgccttctc tgaaaacaat
     3121 gctgagaatg agagcggcgg agacaagccc cccatcgatc ccaataaccc agcagccaac
     3181 tggcttcatg cgcgctccac tcggaaaaag cggtgcccct atacaaaaca ccagaccctg
     3241 gaactggaga aagagtttct gttcaacatg tacctcacca gggaccgcag gtacgaggtg
     3301 gctcgactgc tcaacctcac cgagaggcag gtcaagatct ggttccagaa ccgcaggatg
     3361 aaaatgaaga aaatcaacaa agaccgagca aaagacgagt gaacctgcag aagcttggcc
     3421 gccatggccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3481 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3541 caatgtatct tatcatgtct ggatcgggaa ttaattcggc gcagcaccat ggcctgaaat
     3601 aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa
     3661 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3721 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3781 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3841 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3901 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3961 aggctttttt ggaggcctag gcttttgcaa aaagctgtta acagcttggc actggccgtc
     4021 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4081 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4141 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     4201 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4261 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4321 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4381 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4441 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4501 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4561 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4621 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4681 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4741 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4801 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4861 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4921 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4981 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5041 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5101 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5161 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5221 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5281 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5341 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5401 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5461 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5521 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5581 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5641 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5701 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5761 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5821 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5881 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5941 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6001 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6061 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6121 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6181 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6241 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6301 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6361 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6421 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6481 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6541 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6601 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6661 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6721 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6781 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6841 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6901 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     6961 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     7021 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     7081 ttcacacagg aaacagctat gacatgatta cgaattaa
//
