LOCUS       sequence-313783-        7713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKate tagged MKLP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7713)
  AUTHORS   Rathbun LI, Colicino EG, Manikas J, O'Connell J, Krishnan N, Reilly 
            NS, Coyne S, Erdemci-Tandogan G, Garrastegui A, Freshour J, Santra 
            P, Manning ML, Amack JD, Hehnly H
  TITLE     Cytokinetic bridge triggers de novo lumen formation in vivo.
  JOURNAL   Nat Commun. 2020 Mar 9;11(1):1269. doi: 10.1038/s41467-020-15002-8.
  PUBMED    32152267
REFERENCE   2  (bases 1 to 7713)
  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 
            Commun."; date: "2020-03-9"; volume: "11(1):1269. doi"; pages: " 
            10.1038/s41467-020-15002-8"
COMMENT     Sequence Label: pCS2-mKate-MKLP1
FEATURES             Location/Qualifiers
     source          1..7713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1080..1089
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1086..1781
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     polyA_signal    4743..4877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4767..4786)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4821..4840
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4990..5010)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4990..5008)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5029..5045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5029..5045)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5042..5064)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5053..5069
                     /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(5077..5083,5084..5101,5102..5107))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5122..5143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5260..5277)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5431..6019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5511..5530)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6190..6981,6982..7050))
                     /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     6813..6832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7051..7155)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7181..7636)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7318..7339)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7530..7549
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcagccg gtcgccaccg gtcgccatcg
     1081 ccaccatggt gagcgagctg attaaggaga acatgcacat gaagctgtac atggagggca
     1141 ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca
     1201 cccagaccat gagaatcaag gcggtcgagg gcggccctct ccccttcgcc ttcgacatcc
     1261 tggctaccag cttcatgtac ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg
     1321 acttctttaa gcagtccttc cccgagggct tcacatggga gagagtcacc acatacgaag
     1381 acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca
     1441 acgtcaagat cagaggggtg aacttcccat ccaacggccc tgtgatgcag aagaaaacac
     1501 tcggctggga ggcctccacc gagaccctgt accccgctga cggcggcctg gaaggcagag
     1561 ccgacatggc cctgaagctc gtgggcgggg gccacctgat ctgcaacttg aagaccacat
     1621 acagatccaa gaaacccgct aagaacctca agatgcccgg cgtctactat gtggacagaa
     1681 gactggaaag aatcaaggag gccgacaaag agacctacgt cgagcagcac gaggtggctg
     1741 tggccagata ctgcgacctc cctagcaaac tggggcacag agagctcaag cttcgaattc
     1801 tgcagtcgac aatgaagtca gcgagagcta ggacaccccg gaaacctacc ttgaaaaaag
     1861 ggtcccaaac gaaccttaaa gacccagttg gggtatactg tagggtgcgc ccactgggct
     1921 ttcctgatca agagtgttgc atagaagtga tcaataatac aactgttcag cttcatactc
     1981 ctgagggcta cagactcaac cgaaatggag actataagga gactcagtat tcatttaaac
     2041 aagtatttgg cactcacacc acccagaagg aactctttga tgttgtggct aatcccttgg
     2101 tcaatgacct cattcatggc aaaaatggtc ttctttttac atatggtgtg acgggaagtg
     2161 gaaaaactca cacaatgact ggttctccag aggaaggagg gctgcttcct cgttgtttgg
     2221 acatgatctt taacagtata gggtcatttc aagctaaacg atatgttttc aaatctaatg
     2281 ataggaatag tatggatata cagtgtgagg ttgatgcctt attagaacgt cagaaaagag
     2341 aagctatgcc caatccaaag acttcttcta gcaaacgaca agtagatcca gagtttgcag
     2401 atatgataac tgtacaagaa ttctgcaaag cagaagaggt tgatgaagat agtgtctatg
     2461 gtgtatttgt ctcttatatt gaaatatata ataattacat atatgatcta ttggaagagg
     2521 tgccgtttga tcccataaaa cccaaacctc cacaatctaa attgcttcgt gaagataaga
     2581 accataacat gtatgttgca ggatgtacag aagttgaagt gaaatctact gaggaggctt
     2641 ttgaagtttt ctggagaggc cagaaaaaga gacgtattgc taatacccat ttgaatcgtg
     2701 agtccagccg ttcccatagc gtgttcaaca ttaaattagt tcaggctccc ttggatgcag
     2761 atggagacaa tgtcttacag gaaaaagaac aaatcactat aagtcagttg tccttggtag
     2821 atcttgctgg aagtgaaaga actaaccgga ccagagcaga agggaacaga ttacgtgaag
     2881 ctggtaatat taatcagtca ctaatgacgc taagaacatg tatggatgtc ctaagagagg
     2941 accaaatgta tggaactaac aagatggttc catatcgaga ttcaaagtta acccatctgt
     3001 tcaagaacta ctttgatggg gaaggaaaag tgcggatgat cgtgtgtgtg aaccccaagg
     3061 ctgaagatta tgaagaaaac ttgcaagtca tgagatttgc ggaagtgact caagaagttg
     3121 aagtagcaag acctgtagac aaggcaatat gtggtttaac gcctgggagg agatacagaa
     3181 accagcctcg aggtccagtt ggaaatgaac cattggttac tgacgtggtt ttgcagagtt
     3241 ttccaccttt gccgtcatgc gaaattttgg atatcaacga tgagcagaca cttccaaggc
     3301 tgattgaagc cttagagaaa cgacataact tacgacaaat gatgattgat gagtttaaca
     3361 aacaatctaa tgcttttaaa gctttgttac aagaatttga caatgctgtt ttaagtaaag
     3421 aaaaccacat gcaagggaaa ctaaatgaaa aggagaagat gatctcagga caggaattgg
     3481 aaatagaacg actggaaaag aaaaacaaaa ctttagaata taagattgag attttagaga
     3541 aaacaactac tatctatgag gaagataaac gcaatttgca acaggaactt gaaactcaga
     3601 accagaaact tcagcgacag ttttctgaca aacgcagatt agaagccagg ttgcaaggca
     3661 tggtgacaga aacgacaatg aagtgggaga aagaatgtga gcgtagagtg gcagccaaac
     3721 agctggagat gcaggataaa ctctgggtta aagatgaaaa gctgaaacaa ctgaaggcta
     3781 ttgttactga acctaaaact gagaagccag agagaccctc tcgggagcga gatcgagaaa
     3841 aagttactca aagatctgtt tctccatcac ctgtgcctct ttctagtaac tatattgctc
     3901 agatttccaa cggccagcaa ctcatgagcc agccacagct acataggcgc tctaactctt
     3961 gcagcagcat ttctgtagct tcctgtattt cggaatggga gctgaaaatt cctacgtaca
     4021 acacacctct caaagtcaca tctattgcaa ggcgtaggca gcaggagcca ggacaaagca
     4081 aaacttgtat cgtgtcagac agaaggcgag ggatgtactg gactgaaggc agggaggtgg
     4141 ttcctacatt cagaaatgag atagaaatag aagaggatca ttgcggcagg ttactctttc
     4201 aacctgatca gaacgcacca ccaattcgtc tccgacacag acgatcacgc tctgcaggag
     4261 acagatgggt agatcataag cccgcctcta acatgcaaac tgaaacagtc atgcagccac
     4321 atgtccctca tgccatcaca gtatctgttg caaatgaaaa ggcactagct aagtgtgaga
     4381 agtacatgct gacccaccag gaactagcct ccgatgggga gattgaaact aaactaatta
     4441 agggtgatat ttataaaaca aggggtggtg gacaatctgt tcagtttact gatattgaga
     4501 ctttaaagca agaatcacca aatggtagtc gaaaacgaag atcttccaca gtagcacctg
     4561 cccaaccaga tggtgcagag tctgaatgga ccgatgtaga aacaaggtgt tctgtggctg
     4621 tggagatgag agcaggatcc cagctgggac ctggatatca gcatcacgca caacccaagc
     4681 gcaaaaagcc agtcgacggt accgcggctg cagtctagaa ctatagtgag tcgtattacg
     4741 tagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     4801 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     4861 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     4921 aggtgtggga ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag
     4981 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     5041 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa
     5101 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5161 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5221 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5281 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5341 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5401 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5461 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5521 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5581 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5641 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5701 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5761 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5821 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5881 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5941 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6001 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6061 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6121 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6181 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6241 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6301 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6361 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6421 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6481 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6541 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6601 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6661 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6721 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6781 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6841 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6901 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6961 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7021 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7081 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7141 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     7201 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     7261 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     7321 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     7381 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     7441 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     7501 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     7561 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     7621 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
     7681 gtgcgggcct cttcgctatt acgccagtcg acc
//
