LOCUS       sequence-416602-        7052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of N-terminally mCherry-tagged NIP45 wt 
            resistant to siRNA. Use with Flp-In T-Rex system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7052)
  AUTHORS   Hertz EPT, Vega IA, Kruse T, Wang Y, Hendriks IA, Bizard AH, 
            Eugui-Anta A, Hay RT, Nielsen ML, Nilsson J, Hickson ID, Mailand N
  TITLE     The SUMO-NIP45 pathway processes toxic DNA catenanes to prevent 
            mitotic failure.
  JOURNAL   Nat Struct Mol Biol. 2023 Jul 20. doi: 10.1038/s41594-023-01045-0.
  PUBMED    37474739
REFERENCE   2  (bases 1 to 7052)
  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: "Nat Struct 
            Mol Biol. 2023 Jul 20. doi: 10.1038/s41594-023-01045-0."
COMMENT     Sequence Label: mCherry-NIP45 wt
FEATURES             Location/Qualifiers
     source          1..7052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(649..696)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     828..847
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    980..1204
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1193..1210
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2521..3228)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2796..2815)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3065..3083
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3225..3234
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3321..3345)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3355..3373
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3376..3394
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(3396..3599)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3425..3445)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3600..3979
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4151..4170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4245..4349
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4350..4418,4419..5210)
                     /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(4568..4587)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5381..5969
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5870..5889
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6123..6140
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6257..6278
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6293..6298,6299..6316,6317..6323)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6331..6347
                     /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     6336..6358
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6355..6371
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6355..6371
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    6420..6541
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6431..6450)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6485..6504
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6671..7052,1..641))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(join(6671..7052,1..641))
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc
       61 acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc
      121 gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca
      181 gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat
      241 gggccgaacc cgctcgtctg gctaagatcg gccgcagcga tcgcatccat ggcctccgcg
      301 accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt
      361 gcacggcggg agatgcaata ggtcaggctc tcgctgaatt ccccaatgtc aagcacttcc
      421 ggaatcggga gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca
      481 tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca
      541 cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc
      601 agaaacttct cgacagacgt cgcggtgagt tcaggctttt tggccaagga agttcctata
      661 ctttctagag aataggaact tcggaatagg aacttctagg tacgtgaacc atcaccctaa
      721 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
      781 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
      841 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
      901 cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat accccctaga gccccagctg
      961 gttctttccg cctcagaagc catagagccc accgcatccc cagcatgcct gctattgtct
     1021 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
     1081 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     1141 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     1201 acagtcgagg ctgatcagcg ggtttaaacg ggccctctag actcgagcgg ccgctcagcc
     1261 ccagacctca atgaggtccc cagattccat gcccaggtca gctggcagct ccctgcctga
     1321 aagctttgtc ccatcaaaga agaaggagag cttccgtccc gacagtccca tggcctcctc
     1381 atagtgggac atgagggtct ttagagggga atctcgagac agtgagactt ccagtgtctg
     1441 gtgtttctcc tttccctgca cccggagctg gagctgttgg gacgtctctg tggcctctgg
     1501 agaacttgtt agtaccacac agtcaatgat gtcagccact ccgagcttta gggtcctggg
     1561 agtggcagta ggtgatagct ctgtctctcc aaaaagcaaa aggatcctgc ttggggacac
     1621 cccaaggtgg gtggccatgt ggtccaccac actctgcagg ggctccgaca tcctgagggg
     1681 caatctgacc aggtcagccc ggcaacggat tttgagtggg aagagtcggg gggtctctgg
     1741 gagggtgggc ccttccacca agaccacttc atcctcttgg ccctgttgct cttgaccctg
     1801 aggtggcttg gggctcagac aggaacggag atcctggagg cgcttgttca cctcacttaa
     1861 cttcttgagt gcccgagtat gggttcgaga cttcgttctt ggtgaaggtg gggacagagg
     1921 agagttgtcc agatccagaa actctgtctt tttcttctcc tccttgtctt ttgttcgaag
     1981 ctttgtcttc cagggagagc ctggtgatgg ggagccctca tggtagagac cactcgaatc
     2041 tgccagctct gcctcttcct catcccctgg agggtagagt ttcaggaggg atagatcatc
     2101 aggaatgaga cggagactgc ttttaacctt ccccgagtac accggaacca gcggcgcctc
     2161 ccccggatcc agcaccagcc gccgccgccg cctgaccggc tcccgcgggg gtcctgcggg
     2221 ccgcctgtcc tccccttcgc tgtcactgtt gctgttatcc cgggacgcga ccggccccgg
     2281 gggctccggg ggctccacct caacctcgtc cgcggcaccg cgagcggtgg cgacctccag
     2341 aatttcctca tcgctgtcgg tgaccaagtc cacagacact acgtccagcg tgccccggga
     2401 tggagaccgc tgggcccgag gacgccggcc ccgaccgccc cagccgcccc gaccccctcg
     2461 gccggcaccg ctacctccgg accagcggcc ccgcttcccc acaggctccg ccatggtacc
     2521 cttgtacagc tcgtccatgc cgccggtgga gtggcggccc tcggcgcgtt cgtactgttc
     2581 cacgatggtg tagtcctcgt tgtgggaggt gatgtccaac ttgatgttga cgttgtaggc
     2641 gccgggcagc tgcacgggct tcttggcctt gtaggtggtc ttgacctcag cgtcgtagtg
     2701 gccgccgtcc ttcagcttca gcctctgctt gatctcgccc ttcagggcgc cgtcctcggg
     2761 gtacatccgc tcggaggagg cctcccagcc catggtcttc ttctgcatta cggggccgtc
     2821 ggaggggaag ttggtgccgc gcagcttcac cttgtagatg aactcgccgt cctgcaggga
     2881 ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt
     2941 gaagccctcg gggaaggaca gcttcaagta gtcggggatg tcggcggggt gcttcacgta
     3001 ggccttggag ccgtacatga actgagggga caggatgtcc caggcgaagg gcagggggcc
     3061 acccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc cctcgccctc
     3121 gccctcgatc tcgaactcgt ggccgttcac ggagccctcc atgtgcacct tgaagcgcat
     3181 gaactccttg atgatggcca tgttatcctc ctcgcccttg ctcaccatgg tggcttaagt
     3241 ttaaacgcta gagtccggag gctggatcgg tcccggtgtc ttctatggag gtcaaaacag
     3301 cgtggatggc gtctccaggc gatctgacgg ttcactaaac gagctcgtcg acgatctcta
     3361 tcactgatag ggagatctct atcactgata gggagagctc tgcttatata gacctcccac
     3421 cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag
     3481 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     3541 aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc
     3601 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     3661 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     3721 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     3781 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     3841 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     3901 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     3961 ctagtcaata atcaatgtca acgcgtatat ctggcccgta catcgcgaag cagcgcaaaa
     4021 cgcctaaccc taagcagatt cttcatgcaa ttgtcggtca agccttgcct tgttgtagct
     4081 taaattttgc tcgcgcacta ctcagcgacc tccaacacac aagcagggag cagatactgg
     4141 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatagggg atcgggagat
     4201 ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4261 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4321 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4381 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4441 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4501 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4561 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4621 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4681 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4741 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4801 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4861 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4921 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4981 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5041 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5101 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5161 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5221 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5281 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5341 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5401 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5461 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5521 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5581 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5641 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5701 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5761 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5821 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5881 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5941 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6001 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6061 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6121 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     6181 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     6241 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     6301 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     6361 acagctatga ccatgattac gccaagctct agctagaggt cgacggtata cagacatgat
     6421 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     6481 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     6541 tggggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt
     6601 cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc
     6661 gtagtacgtg ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg
     6721 cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac
     6781 gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg
     6841 catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca
     6901 tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg
     6961 tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt
     7021 gggaatcccc gaacatcgcc tcgctccagt ca
//
