LOCUS       sequence-474381-        6046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For RNA injection of Nanog-mNG into Zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6046)
  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 6046)
  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: Nanog-mNG
FEATURES             Location/Qualifiers
     source          1..6046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1143..1277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1180..1199)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1234..1253
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1310..1336)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1343..1369)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1388..1411)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3322..3340)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3323..3340)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(3375..4359)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      4430..4885
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4517..4536)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4727..4748
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4911..5015
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5016..5084,5085..5876)
                     /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(5234..5253)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tgggtaccgg gcccaatgca ttggcgccgc ggccgcgaat
     1081 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1141 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1201 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1261 cttatcatgt ctggatctac gtaatacgac tcactatagt tctagattaa gcgtagtctg
     1321 gcacgtcgta ggggtaagct ccagcgtagt ctggcacgtc gtaggggtaa ctcgagaggc
     1381 cttgttactt atcgtcatcg tctttataat ccttgtacag ctcgtccatt cccatcacgt
     1441 ctgtgaaagc cttctgccac tccttgaagt tcagctctgt cttgctgtgc ttcagctctg
     1501 tctttctgaa cacgtacata ggctggttct tcaggtagtt agcagccata ggcttagcga
     1561 atgtgtatgt tgttctagct gtgcttctgt atctctttcc gtttcctgtt gtgtagctcc
     1621 acttgaatgt gctgatgatt gtcttgtcgt tagggtatgt cttcttgctt ctacaccagt
     1681 cagcagctgt caggctgttt gtcatcacag gtccgtcagc agggaatcct gttcccttca
     1741 cctgagcctc tcccttgatg tggcttccct cgtatgtgta tctgtagttc actgtcaggc
     1801 tagctccgtc ctcgaactgc attgttctgt gcacctggta tccgcttccg tccaccatag
     1861 cagcctggaa agggctcatt ccgtcagggt aaggcaggta ctggtggaat ccgtatccga
     1921 tgtgaggcac caggatccaa gggctgaact gcaggtctcc ctttgtgctc ttcaggttca
     1981 gctcctcgta tccgtcgtta gggtttcctg ttccctgtcc caccatgtcg aagtccactc
     2041 cgttgatgct tccgaagatg tgcagctcgt gtgtagcagg caggctagcc atgttgtcct
     2101 cctctccctt gctcaccatt cctcccccgc tcagcaaagt tattccttta gttgcccaca
     2161 ggccagcagt aggcgaagtt gccagcggag aagggctgct ttctttgttc actgcattgc
     2221 tgccatcacg tgctggcatg tagctgatgg ggttggggaa ctcatagtgc gtcacagctg
     2281 ggggcagagg ccaggttcct gggggtctcg aggtggcctg tgttggggag cgtggctgtg
     2341 ggtaactggg gtagaagggt gtttttggag gactcctctt gaaaacaggt tctctcacct
     2401 gagggttaat gtagtggtct tggtttgctc cagggggttc actctgaaac tgagattgag
     2461 aagctggtgt gttgggtaca gcattgacaa catatctctc agtcatccaa ctgctgtctc
     2521 tctgatgcct cttgagtttc atcctgcggt tctgaaacca tgtttttacc tgcttgtatg
     2581 ttaatcctgt tgctccagca agcgttttca tctctgcagg cgtgaggtat ctctggacat
     2641 taaaccgatt gaccaaagca ttcatctgtt cttcagaaaa agcagcccgg gtcttgcggg
     2701 gcagggtcgg aggccggaca ggagatgcag tcaatgcagg cagaggaggc tgggtatcga
     2761 cccccagatt cagattgact ggctcctctc ctgctgtgga ggttaagttc cgttctccac
     2821 tgtcaggact accactgtct gtttcatagt ctcgatctct ccaggatggc agattgaggt
     2881 tggtcagagg aaccccttct cgactgctcg ctgagctcca ggaatctggc gtgtgggcct
     2941 cggaatcact gggtgtatca ctgctggtct gctctgcatc tttggccttt tggtcgggct
     3001 ctgtcttgtt gtaggacaga aagagccgtc cgttctgcga gtgtcccggg tactggctgt
     3061 aggtgctgct ggcgccgccg ttctccgggc tccagggtgg cgactgtgca gtttgagcgg
     3121 taaagtagcc tgctgtgacc ccgccggagt gtgtatacgc ggcatcgggc cagctcagat
     3181 tcgggacgcc gtgctctgac acttgcgggt acatgagccc gtatgcatat gcatgataag
     3241 atgggttaaa attgtaactc actggcatct tccagtccgc catatcgatg ggatcctgca
     3301 aaaagaacaa gtagcttgta ttctatagtg tcacctaaat caagcttgct ccgacgtccc
     3361 caggcagaat ggcggttccc taaacgagca ttgcttatat agacctccca ttaggcacgc
     3421 ctaccgccca tttacgtcaa tggaacgccc atttgcgtca ttgcccctcc ccattgacgt
     3481 caatggggat gtacttggca gccatcgcgg gccatttacc gccattgacg tcaatgggag
     3541 tactgccaat gtaccctggc gtacttccaa tagtaatgta cttgccaagt tactattaat
     3601 agatattgat gtactgccaa gtgggccatt taccgtcatt gacgtcaata gggggcgtga
     3661 gaacggatat gaatgggcaa tgagccatcc cattgacgtc aatggtgggt ggtcctattg
     3721 acgtcaatgg gcattgagcc aggcgggcca tttaccgtaa ttgacgtcaa tgggggaggc
     3781 gccatatacg tcaataggac cgcccatatg acgtcaatag gaaagaccat atatagagac
     3841 cattgacgtc aatgggggag tggctatggg cggtattagg aagccccata tatggtatat
     3901 gggaccgccc attgggaggg gctatctacg tcaataggaa aacccatata tggaatacta
     3961 tatggcatag ggccaataca tagtattgaa cctggccaat agccatattg gcatagggcc
     4021 atattggata ttgcctatat attgatcctg gcatatagcc aatatggccg ccattattgg
     4081 caccatgcca attcaatatg gcggacttgg cacagggcca agacccctcc tcaaacttgg
     4141 caccgtgcca agtagacccc tccccagttg gcacagtgcc aggtccgcca tattgaattg
     4201 gcatggtgcc aagtccgcca tattgaattg gctggggtcc agatccacca tattgaattg
     4261 gcacgagtcc atatacgcca tattgaattg gcacaggtcc agatccacca tattgaattg
     4321 gcatgagtcc atatacgcca tattgaattg gcggtcgact ggcgtaatag cgaagaggcc
     4381 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     4441 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4501 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4561 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4621 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4681 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4741 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4801 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4861 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4921 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4981 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     5041 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5101 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5161 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5221 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5281 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5341 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5401 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5461 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5521 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5581 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5641 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5701 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5761 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5821 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5881 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5941 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6001 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
