LOCUS       sequence-393557-        5366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 551, Emission:
            565.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5366)
  AUTHORS   Sakaue-Sawano A, Kurokawa H, Morimura T, Hanyu A, Hama H, Osawa H, 
            Kashiwagi S, Fukami K, Miyata T, Miyoshi H, Imamura T, Ogawa M, 
            Masai H, Miyawaki A
  TITLE     Visualizing spatiotemporal dynamics of multicellular cell-cycle 
            progression.
  JOURNAL   Cell. 2008 Feb 8;132(3):487-98. doi: 10.1016/j.cell.2007.12.033.
  PUBMED    18267078
REFERENCE   2  (bases 1 to 5366)
  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: 
            "10.1016/j.cell.2007.12"; journalName: "Cell"; date: "2008-02-8- 8";
            volume: "132"; issue: "3"; pages: "487-98"
COMMENT     Sequence Label: mKO2-pBAD
FEATURES             Location/Qualifiers
     source          1..5366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     219..238
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     472..489
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    504..644
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(570..589)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     730..752
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     890..909
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(1089..1967)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1994..2278
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2201..2220
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             2305..2327
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2347..2364
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2368..2400
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             2413..2433
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2440..3096
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     primer_bind     complement(3142..3159)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3312..3398
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3490..3517
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3536..3627
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3628..3696,3697..4488)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3846..3865)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4530..4985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4617..4636)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4827..4848
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      join(5096..5366,1..318)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
       61 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      121 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      181 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      241 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      301 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      361 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
      421 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
      481 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
      541 atttcacacc gcatctggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      601 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
      661 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
      721 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
      781 ggcagaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
      841 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
      901 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
      961 cgtagaggat ctgctcatgt ttgacagctt atcatcgatg cataatgtgc ctgtcaaatg
     1021 gacgaagcag ggattctgca aaccctatgc tactccgtca agccgtcaat tgtctgattc
     1081 gttaccaatt atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg
     1141 aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg
     1201 tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc
     1261 ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg
     1321 cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata
     1381 tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta
     1441 tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca
     1501 agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt
     1561 tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta
     1621 ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa
     1681 acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct
     1741 ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc
     1801 accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg
     1861 ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca
     1921 ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc
     1981 ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc
     2041 ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa
     2101 caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga
     2161 aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc
     2221 ataagattag cggatactac ctgacgcttt ttatcgcaac tctctactgt ttctccatac
     2281 ccgttttttt gggctagaaa taattttgtt taactttaag aaggagatat acatatgcgg
     2341 ggttctcatc atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt
     2401 cgggatctgt acgagaacct gtacttccag ggctcgagca tggtgagtgt gattaaacca
     2461 gagatgaaga tgaggtacta catggacggc tccgtcaatg ggcatgagtt cacaattgaa
     2521 ggtgaaggca caggcagacc ttacgaggga catcaagaga tgacactacg cgtcacaatg
     2581 gccgagggcg ggccaatgcc tttcgcgttt gacttagtgt cacacgtgtt ctgttacggc
     2641 cacagagtat ttactaaata tccagaagag ataccagact atttcaaaca agcatttcct
     2701 gaaggcctgt catgggaaag gtcgttggag ttcgaagatg gtgggtccgc ttcagtcagt
     2761 gcgcatataa gccttagagg aaacaccttc taccacaaat ccaaatttac tggggttaac
     2821 tttcctgccg atggtcctat catgcaaaac caaagtgttg attgggagcc atcaaccgag
     2881 aaaattactg ccagcgacgg agttctgaag ggtgatgtta cgatgtacct aaaacttgaa
     2941 ggaggcggca atcacaaatg ccaaatgaag actacttaca aggcggcaaa agagattctt
     3001 gaaatgccag gagaccatta catcggccat cgcctcgtca ggaaaaccga aggcaacatt
     3061 actgagcagg tagaagatgc agtagctcat agctaagaat tcgaagcttg gctgttttgg
     3121 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     3181 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     3241 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     3301 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     3361 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     3421 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     3481 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     3541 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3601 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3661 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3721 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3781 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3841 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     3901 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3961 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4021 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4081 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4141 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     4201 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4261 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4321 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4381 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4441 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4501 gtttactcat atatacttta gattgattta cgcgccctgt agcggcgcat taagcgcggc
     4561 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4621 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4681 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4741 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4801 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4861 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4921 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4981 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5041 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5101 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5161 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5221 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     5281 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5341 gtggcgataa gtcgtgtctt accggg
//
