LOCUS       sequence-391819-        5844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  visualization of cleavage furrow in living mammalian mitotic cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5844)
  AUTHORS   Tanaka S, Senda-Murata K, Kawakita-Yamaguchi A, Fukada T, Sugimoto K
  TITLE     Live cell imaging of anaphase bridge formation and the subsequent 
            cleavage furrow regression induced by the Aurora B kinase inhibitor 
            AZD1152-HQPA
  JOURNAL   Bioimages 25: 7-19, 2017
REFERENCE   2  (bases 1 to 5844)
  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: "Bioimages 
            25: 7-19, 2017"
COMMENT     Sequence Label: pmOrange 2-Actin
FEATURES             Location/Qualifiers
     source          1..5844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      598..607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             604..1311
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    2456..2508
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2632..2753
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2669..2688)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2723..2742
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2760..3215)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2897..2918)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3109..3128
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3242..3346
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3348..3705
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3348..3539
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3371..3391)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3556..3691
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3740..4534
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3740..4534
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3794..3813)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4404..4423
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4722..4741)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4766..4813
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5142..5730
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5631..5650
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccgcc
      601 accatggtga gcaagggcga ggagaataac atggccatca tcaaggagtt catgcgcttc
      661 aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
      721 ggccgcccct acgagggctt tcagaccgct aagctgaagg tgaccaaggg tggccccctg
      781 cccttcgcct gggacatcct gtcccctcat ttcacctacg gctccaaggc ctacgtgaag
      841 caccccgccg acatccccga ctacttcaag ctgtccttcc ccgagggctt caagtgggag
      901 cgcgtgatga actacgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
      961 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1021 gtgatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     1081 ggtgccctga agggcaagat caagatgagg ctgaagctga aggacggcgg ccactacacc
     1141 tccgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacatc
     1201 gtcgacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1261 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gagatctcga
     1321 gctatggatg atgatatcgc cgcgctcgtc gtcgacaacg gctccggcat gtgcaaggcc
     1381 ggcttcgcgg gcgacgatgc cccccgggcc gtcttcccct ccatcgtggg gcgccccagg
     1441 caccagggcg tgatggtggg catgggtcag aaggattcct atgtgggcga cgaggcccag
     1501 agcaagagag gcatcctcac cctgaagtac cccatcgagc acggcatcgt caccaactgg
     1561 gacgacatgg agaaaatctg gcaccacacc ttctacaatg agctgcgtgt ggctcccgag
     1621 gagcaccccg tgctgctgac cgaggccccc ctgaacccca aggccaaccg cgagaagatg
     1681 acccagatca tgtttgagac cttcaacacc ccagccatgt acgttgctat ccaggctgtg
     1741 ctatccctgt acgcctctgg ccgtaccact ggcatcgtga tggactccgg tgacggggtc
     1801 acccacactg tgcccatcta cgaggggtat gccctccccc atgccatcct gcgtctggac
     1861 ctggctggcc gggacctgac tgactacctc atgaagatcc tcaccgagcg cggctacagc
     1921 ttcaccacca cggccgagcg ggaaatcgtg cgtgacatta aggagaagct gtgctacgtc
     1981 gccctggact tcgagcaaga gatggccacg gctgcttcca gctcctccct ggagaagagc
     2041 tacgagctgc ctgacggcca ggtcatcacc attggcaatg agcggttccg ctgccctgag
     2101 gcactcttcc agccttcctt cctgggcatg gagtcctgtg gcatccacga aactaccttc
     2161 aactccatca tgaagtgtga cgtggacatc cgcaaagacc tgtacgccaa cacagtgctg
     2221 tctggcggca ccaccatgta ccctggcatt gccgacagga tgcagaagga gatcactgcc
     2281 ctggcaccca gcacaatgaa gatcaagatc attgctcctc ctgagcgcaa gtactccgtg
     2341 tggatcggcg gctccatcct ggcctcgctg tccaccttcc agcagatgtg gatcagcaag
     2401 caggagtatg acgagtccgg cccctccatc gtccaccgca aatgcttcta gggatctcga
     2461 gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga
     2521 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2581 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2641 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2701 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
     2761 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2821 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2881 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2941 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3001 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3061 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3121 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3181 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3241 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3301 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3361 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3421 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3481 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3541 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3601 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3661 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3721 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3781 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3841 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3901 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3961 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4021 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4081 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4141 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4201 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4261 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4321 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4381 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4441 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4501 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4561 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4621 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4681 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4741 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4801 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4861 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4921 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     4981 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5041 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5101 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5161 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5221 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5281 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5341 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5401 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5461 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5521 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5581 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5641 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5701 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5761 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5821 gataaccgta ttaccgccat gcat
//
