LOCUS       sequence-383840-        7782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mNeonGreen fused to p80-Katanin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7782)
  AUTHORS   Schneider MWG, Gibson BA, Otsuka S, Spicer MFD, Petrovic M, Blaukopf
            C, Langer CCH, Batty P, Nagaraju T, Doolittle LK, Rosen MK, Gerlich 
            DW
  TITLE     A mitotic chromatin phase transition prevents perforation by 
            microtubules.
  JOURNAL   Nature. 2022 Sep;609(7925):183-190. doi: 10.1038/s41586-022-05027-y.
            Epub 2022 Aug 3.
  PUBMED    35922507
REFERENCE   2  (bases 1 to 7782)
  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: 
            "10.1038/s41586-022-05027-y"; journalName: "Nature"; date: 
            "2022-09"; volume: "609"; issue: "7925"; pages: "183-190"
COMMENT     Sequence Label: p80-Katanin-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..7782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(102..893,894..962))
                     /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     725..744
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(963..1067)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1142..1161)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1333..1712
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1713..1916
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1867..1887
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1945..1954
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3872..3881
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3946..4656
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      4201..4210
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          4725..4954
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     4937..4956
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(5026..5600,5601..5603,5604..5612)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5193..5210)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5420..5439
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5632..6231
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5632..5651)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6128..6148
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6367..6384)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6373..6597
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6644..6663
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6723..6739)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6723..6739)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6736..6758)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6747..6763
                     /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)."
     promoter        complement(join(6771..6777,6778..6795,6796..6801))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6816..6837
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6954..6971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7125..7713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7205..7224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
       61 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      121 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
      181 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
      241 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
      301 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
      361 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
      421 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
      481 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
      541 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
      601 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
      661 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
      721 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
      781 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
      841 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
      901 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
      961 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1021 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1081 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     1141 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     1201 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     1261 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
     1321 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     1381 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     1441 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     1501 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     1561 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     1621 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     1681 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     1741 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     1801 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     1861 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt
     1921 tagtgaaccg tcagatccgc tagcgccacc atggccaccc ctgtggtcac caagacagcc
     1981 tggaagttgc aagagatcgt cgcccatgcc agcaatgtgt cctccttggt gctgggcaag
     2041 gcctcagggc ggctgctggc cactggtggg gatgactgcc gagtcaacct gtggtccata
     2101 aacaagccta actgcatcat gagcctgaca ggtcacacgt cccctgtgga gagtgtccgt
     2161 ctcaacactc ctgaggagct catcgtagct ggttcccagt cgggctccat ccgcgtctgg
     2221 gacctggaag ctgctaaaat tcttcgaaca ctcatgggcc acaaagccaa catctgcagc
     2281 ctggatttcc acccctatgg tgaatttgtg gcctcaggct cccaggacac aaacattaag
     2341 ctctgggaca tcaggaggaa aggctgtgtc ttccgatata gggggcacag ccaggctgtg
     2401 cggtgtctcc ggttcagccc tgatgggaag tggctggcgt cggcagcaga tgaccacaca
     2461 gtgaagcttt gggatctgac ggctggcaag atgatgtccg agttcccggg tcatacaggg
     2521 cctgtcaatg tggtggagtt tcaccccaat gaatatctcc tagcttctgg cagctccgac
     2581 aggacaatcc gcttctggga cctggagaag ttccaggtgg tgagctgcat tgaaggggag
     2641 cccgggcccg tcaggagtgt cctcttcaac cccgacggct gctgcttgta cagcggctgc
     2701 caggactcgc tgcgtgtcta cggctgggaa cctgagcgct gctttgatgt ggtccttgtc
     2761 aactggggca aggtggctga cctggccatc tgcaatgacc agctgatagg tgtggctttc
     2821 tcccagagta atgtctcctc ttatgtggtg gacctgacga gagtcaccag gacgggaact
     2881 gtgacccagg accctgtgca ggctaaccag cctctgacac aacagacccc taaccctggt
     2941 gtctccctcc gccgcatcta tgagcggccc agcacgacat gcagcaagcc tcagagggta
     3001 aagcacaact cagagagcga gcggcgcagc cccagcagcg aagatgacag ggacgagcga
     3061 gagtcccggg cggagatcca gaacgcggag gactacaatg agattttcca gcccaagaac
     3121 agcatcagtc ggacaccacc ccggagaagt gagcccttcc cagcaccccc agaagatgat
     3181 gcagccacag tgaaggaggt ctcaaaaccc agcccagcca tggatgtgca gttgccgcag
     3241 ttgccggtgc caaatctcga ggtcccagca cgaccgtcag tcatgacttc cactcctgca
     3301 cccaagggtg aacctgacat catccctgcc acacggaatg agcccattgg cttaaaggcc
     3361 tcggacttct tgcccgccgt gaaggtcccc cagcaggcag agctggtgga cgaggatgcc
     3421 atgtcgcaga tacgcaaagg ccacgacacc atgtttgtgg tgctcaccag ccgccacaag
     3481 aacctggaca ctgtccgagc tgtatggact acaggagata tcaagacatc agtggactct
     3541 gcagtggcca tcaatgacct gtctgtggta gtggacctcc tgaacattgt caatcagaaa
     3601 gcctccctgt ggaagctgga cctgtgcacc acagtgctgc cccagatcga gaaactgctg
     3661 cagagcaagt atgagagcta tgtccagacc gggtgcacct ccctgaagct gatcctgcag
     3721 cggttcctac ccctcatcac tgacattttg gcagccccgc cctctgtggg tgtggacatc
     3781 agccgagagg agaggttgca caagtgtcga ctctgcttca aacaactcaa gagcatcagt
     3841 ggcctcgtca agagcaagtc gggtttgagt ggccgccatg gcagtgcctt ccgtgagctg
     3901 cacctactca tggccagttt ggaccccgtc gccaccggat cctctatggt gagcaagggc
     3961 gaggaggata acatggcctc tctcccagcg acacatgagt tacacatctt tggctccatc
     4021 aacggtgtgg actttgacat ggtgggtcag ggcaccggca atccaaatga tggttatgag
     4081 gagttaaacc tgaagtccac caagggtgac ctccagttct ccccctggat tctggtccct
     4141 catatcgggt atggcttcca tcagtacctg ccctaccctg acgggatgtc gcctttccag
     4201 gccgccatgg tagatggctc cggataccaa gtccatcgca caatgcagtt tgaagatggt
     4261 gcctccctta ctgttaacta ccgctacacc tacgagggaa gccacatcaa aggagaggcc
     4321 caggtgaagg ggactggttt ccctgctgac ggtcctgtga tgaccaactc gctgaccgct
     4381 gcggactggt gcaggtcgaa gaagacttac cccaacgaca aaaccatcat cagtaccttt
     4441 aagtggagtt acaccactgg aaatggcaag cgctaccgga gcactgcgcg gaccacctac
     4501 acctttgcca agccaatggc ggctaactat ctgaagaacc agccgatgta cgtgttccgt
     4561 aagacggagc tcaagcactc caagaccgag ctcaacttca aggagtggca aaaggccttt
     4621 accgatgtga tgggcatgga cgagctgtac aagtaagcgg ccgcatagat aactgatcca
     4681 gtgtgctgga attaattcgc tgtctgcgag ggccagctgt tggggtgagt actccctctc
     4741 aaaagcgggc atgacttctg cgctaagatt gtcagtttcc aaaaacgagg aggatttgat
     4801 attcacctgg cccgcggtga tgcctttgag ggtggccgcg tccatctggt cagaaaagac
     4861 aatctttttg ttgtcaagct tgaggtgtgg caggcttgag atctggccat acacttgagt
     4921 gacaatgaca tccactttgc ctttctctcc acaggtgtcc actcccaggt ccaactgcag
     4981 gtcgagcatg catctagggc ggccaattcc gcccctctcc ccccccccct ctccctcccc
     5041 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     5101 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     5161 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     5221 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     5281 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     5341 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     5401 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     5461 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     5521 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     5581 ttgaaaaaca cgatgataag cttgccacaa cccacaagga gacgaccttc catgaccgag
     5641 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     5701 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     5761 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     5821 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     5881 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     5941 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     6001 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     6061 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     6121 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     6181 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc
     6241 cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atggctccga ccgaagccac
     6301 ccggggcggc cccgccgacc ccgcacccgc ccccgaggcc caccgactct agagctcgct
     6361 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     6421 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     6481 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     6541 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     6601 ctgaggcgga aagaaccagc tggggctcga gtgcattcta gttgtggttt gtccaaactc
     6661 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     6721 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6781 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6841 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6901 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6961 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7021 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7081 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7141 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7201 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7261 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7321 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7381 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7441 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7501 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7561 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7621 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7681 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7741 tctgacgctc agtggaacga aaactcacgt taagggattt tg
//
