LOCUS       sequence-313780-        8436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry tagged MKLP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8436)
  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 8436)
  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-mCherry-MKLP1
FEATURES             Location/Qualifiers
     source          1..8436
                     /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..1793
                     /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"
     regulatory      1794..1803
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1800..2507
                     /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"
     polyA_signal    5466..5600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5490..5509)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5544..5563
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5713..5733)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5713..5731)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5752..5768)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5752..5768)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5765..5787)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5776..5792
                     /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(5800..5806,5807..5824,5825..5830))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5845..5866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5983..6000)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6154..6742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6234..6253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6913..7704,7705..7773))
                     /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     7536..7555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7774..7878)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7904..8359)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8041..8062)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8253..8272
                     /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 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
     1141 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     1201 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     1261 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     1321 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     1381 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     1441 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     1501 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     1561 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     1621 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     1681 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     1741 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aaggccacca
     1801 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     1861 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1921 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     1981 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     2041 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     2101 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     2161 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     2221 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     2281 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     2341 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     2401 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     2461 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagctc aagcttcgaa
     2521 ttctgcagtc gacaatgaag tcagcgagag ctaggacacc ccggaaacct accttgaaaa
     2581 aagggtccca aacgaacctt aaagacccag ttggggtata ctgtagggtg cgcccactgg
     2641 gctttcctga tcaagagtgt tgcatagaag tgatcaataa tacaactgtt cagcttcata
     2701 ctcctgaggg ctacagactc aaccgaaatg gagactataa ggagactcag tattcattta
     2761 aacaagtatt tggcactcac accacccaga aggaactctt tgatgttgtg gctaatccct
     2821 tggtcaatga cctcattcat ggcaaaaatg gtcttctttt tacatatggt gtgacgggaa
     2881 gtggaaaaac tcacacaatg actggttctc cagaggaagg agggctgctt cctcgttgtt
     2941 tggacatgat ctttaacagt atagggtcat ttcaagctaa acgatatgtt ttcaaatcta
     3001 atgataggaa tagtatggat atacagtgtg aggttgatgc cttattagaa cgtcagaaaa
     3061 gagaagctat gcccaatcca aagacttctt ctagcaaacg acaagtagat ccagagtttg
     3121 cagatatgat aactgtacaa gaattctgca aagcagaaga ggttgatgaa gatagtgtct
     3181 atggtgtatt tgtctcttat attgaaatat ataataatta catatatgat ctattggaag
     3241 aggtgccgtt tgatcccata aaacccaaac ctccacaatc taaattgctt cgtgaagata
     3301 agaaccataa catgtatgtt gcaggatgta cagaagttga agtgaaatct actgaggagg
     3361 cttttgaagt tttctggaga ggccagaaaa agagacgtat tgctaatacc catttgaatc
     3421 gtgagtccag ccgttcccat agcgtgttca acattaaatt agttcaggct cccttggatg
     3481 cagatggaga caatgtctta caggaaaaag aacaaatcac tataagtcag ttgtccttgg
     3541 tagatcttgc tggaagtgaa agaactaacc ggaccagagc agaagggaac agattacgtg
     3601 aagctggtaa tattaatcag tcactaatga cgctaagaac atgtatggat gtcctaagag
     3661 aggaccaaat gtatggaact aacaagatgg ttccatatcg agattcaaag ttaacccatc
     3721 tgttcaagaa ctactttgat ggggaaggaa aagtgcggat gatcgtgtgt gtgaacccca
     3781 aggctgaaga ttatgaagaa aacttgcaag tcatgagatt tgcggaagtg actcaagaag
     3841 ttgaagtagc aagacctgta gacaaggcaa tatgtggttt aacgcctggg aggagataca
     3901 gaaaccagcc tcgaggtcca gttggaaatg aaccattggt tactgacgtg gttttgcaga
     3961 gttttccacc tttgccgtca tgcgaaattt tggatatcaa cgatgagcag acacttccaa
     4021 ggctgattga agccttagag aaacgacata acttacgaca aatgatgatt gatgagttta
     4081 acaaacaatc taatgctttt aaagctttgt tacaagaatt tgacaatgct gttttaagta
     4141 aagaaaacca catgcaaggg aaactaaatg aaaaggagaa gatgatctca ggacaggaat
     4201 tggaaataga acgactggaa aagaaaaaca aaactttaga atataagatt gagattttag
     4261 agaaaacaac tactatctat gaggaagata aacgcaattt gcaacaggaa cttgaaactc
     4321 agaaccagaa acttcagcga cagttttctg acaaacgcag attagaagcc aggttgcaag
     4381 gcatggtgac agaaacgaca atgaagtggg agaaagaatg tgagcgtaga gtggcagcca
     4441 aacagctgga gatgcaggat aaactctggg ttaaagatga aaagctgaaa caactgaagg
     4501 ctattgttac tgaacctaaa actgagaagc cagagagacc ctctcgggag cgagatcgag
     4561 aaaaagttac tcaaagatct gtttctccat cacctgtgcc tctttctagt aactatattg
     4621 ctcagatttc caacggccag caactcatga gccagccaca gctacatagg cgctctaact
     4681 cttgcagcag catttctgta gcttcctgta tttcggaatg ggagctgaaa attcctacgt
     4741 acaacacacc tctcaaagtc acatctattg caaggcgtag gcagcaggag ccaggacaaa
     4801 gcaaaacttg tatcgtgtca gacagaaggc gagggatgta ctggactgaa ggcagggagg
     4861 tggttcctac attcagaaat gagatagaaa tagaagagga tcattgcggc aggttactct
     4921 ttcaacctga tcagaacgca ccaccaattc gtctccgaca cagacgatca cgctctgcag
     4981 gagacagatg ggtagatcat aagcccgcct ctaacatgca aactgaaaca gtcatgcagc
     5041 cacatgtccc tcatgccatc acagtatctg ttgcaaatga aaaggcacta gctaagtgtg
     5101 agaagtacat gctgacccac caggaactag cctccgatgg ggagattgaa actaaactaa
     5161 ttaagggtga tatttataaa acaaggggtg gtggacaatc tgttcagttt actgatattg
     5221 agactttaaa gcaagaatca ccaaatggta gtcgaaaacg aagatcttcc acagtagcac
     5281 ctgcccaacc agatggtgca gagtctgaat ggaccgatgt agaaacaagg tgttctgtgg
     5341 ctgtggagat gagagcagga tcccagctgg gacctggata tcagcatcac gcacaaccca
     5401 agcgcaaaaa gccagtcgac ggtaccgcgg ctgcagtcta gaactatagt gagtcgtatt
     5461 acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     5521 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     5581 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     5641 gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc
     5701 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     5761 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat
     5821 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5881 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5941 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6001 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6061 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6661 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6841 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6901 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7021 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7081 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7441 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7501 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7561 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7621 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7681 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7741 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7801 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     7921 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     7981 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     8041 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     8101 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     8161 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     8221 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     8281 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     8341 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     8401 tcggtgcggg cctcttcgct attacgccag tcgacc
//
