LOCUS       sequence-258972-        5605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pClimDC-H2B:mV nuclear marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5605)
  AUTHORS   Kozyczkowska A, Najle SR, Ocana-Pallares E, Areste C, Shabardina V, 
            Ara PS, Ruiz-Trillo I, Casacuberta E
  TITLE     Stable transfection in protist Corallochytrium limacisporum 
            identifies novel cellular features among unicellular animals 
            relatives.
  JOURNAL   Curr Biol. 2021 Sep 27;31(18):4104-4110.e5. doi: 
            10.1016/j.cub.2021.06.061. Epub 2021 Jul 21.
  PUBMED    34293333
REFERENCE   2  (bases 1 to 5605)
  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.cub.2021.06.061"; journalName: "Curr Biol"; date: 
            "2021-09-27- 27"; volume: "31"; issue: "18"; pages: "4104-4110.e5"
COMMENT     Sequence Label: Clim-DC2-H2B2-Venus
FEATURES             Location/Qualifiers
     source          1..5605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      934..943
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(940..942,943..945,946..1656)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDRMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(985..1006)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1246..1265)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1593..1614
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1685..1806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1843..1862
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(1991..2059,2060..2851)
                     /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     complement(2209..2228)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3022..3610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3511..3530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3764..3781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3898..3919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3934..3939,3940..3957,3958..3964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3972..3988
                     /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)."
     primer_bind     3977..3999
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3996..4012
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3996..4012
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4801..5400
                     /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(4801..4820)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5297..5317
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5423..5544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggattcatc gactgtggcc ggcgaaatgc tgggcgcctt gaaactgaac ctgttcggat
       61 ccgggttgcg cgcttccatt gttttggttt ccgtgtctgc aaggcgggtc tggacgtgac
      121 gctttggggg agagggagag cagagataca attctagcca cgaatttcag ccaatagtca
      181 cagaggggag ttgcgatcct gcgaattggc ttaggtctgt gaggtctttc acggtttcga
      241 aagcagcgca gaggggagac cgactaccaa aagccataac actccaccga gatatccgca
      301 ctcttcgcta gatgagcttg cttctcctct tgtcacaaat cagcccaatg atagtcactg
      361 ttcaacaacg gtgcagtgta aatattcgtt gtgcgtcctt gatgagcaga aggtacgaga
      421 agcaggaaat catttcataa tggaaaacca ttcactgtct ttctgtattg cgaaaagagt
      481 gatcataggt aatcgcaacg gcgggtcttt agaacagtca cataccagat caagcttgtc
      541 gacatgccac ctgttgcatc caagaagggc gagaagaagg ccgccggcaa gaagggcgga
      601 gacaagggcg gtaagcgcaa gaaggcccgc aaggaatctt actccatcta catctacaag
      661 gttctcaagc aggtccaccc tgacactggt gtctccaaca aagccatgtt gatcatgaac
      721 tccttcgtgt ctgacatttt cgagcgtgtc gccaacgaag cttcccgtct tgccaagtac
      781 aacaagaagt ccaccatcac ttcccgggag attcagactg ctgtccgtct gctcctgcct
      841 ggtgagcttg ctaagcacgc cgtctcagaa ggtaccaagg ccgtgaccaa gtacacttcc
      901 tctaagaacg cggccgcatc catcacactg gcggccgcca tggtgagcaa gggcgaggag
      961 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1021 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
     1081 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
     1141 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1201 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1261 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1321 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1381 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     1441 atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     1501 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     1561 ctgagcaaag accccaacga gaagcgcgat cgcatggtcc tgctggagtt cgtgaccgcc
     1621 gccgggatca ctctcggcat ggacgagctg tacaagctcg agtaagctag cagatctagg
     1681 gccctaagat acattgatga gtttggacaa accacaacaa gaatgcagtg aaaaaaatgc
     1741 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1801 caagttgccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     1861 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     1921 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaattgaaa
     1981 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2041 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2101 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2161 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2221 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2281 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2341 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2401 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2461 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2521 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2581 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2641 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2701 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2761 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2821 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2881 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2941 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3001 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3061 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3121 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3181 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3241 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3301 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3361 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3421 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3481 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3541 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3601 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3661 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3721 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3781 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     3841 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     3901 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     3961 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     4021 cgccaagctt atcggtggga ctgtgagaac tgctacctgc ctgtgcagag gagagcacac
     4081 tacattcacc attgaaaata tggcccactg aactgatcaa gccaagtgaa gactgcgcgc
     4141 ctgtgttctg attcttccca tggtacaagt actgaaactt tggaagcgcc agagaattca
     4201 cagacgcaca gaagacagcg catggtggag gctgtgtgtg ttcaccgagg tgtcctctgc
     4261 ggaaggtgcc gttcattatg tattcggagt aaagcacaac ctcttcaatg ctgctccctt
     4321 ccttcgcttg tttcttggat agttatcgtc tgtcagaatc tttcggtaca ttcagtatag
     4381 agttcgcgat cggtagcaac agtcggacgg accctgtcag gacggtgtat tcggtggcct
     4441 acttttcttt tgttgcatag ctgttcctga cgcgatgcgc caagcgccgt catattgtgc
     4501 tgtgtgcccc tatgtggagc aggcatagag ctcttgccaa atgttgggaa gagctgttgg
     4561 agaagcgaca gagcagtgcc aagacgacga acgaggtgtg cgagctgtag tgctagtgct
     4621 tcctaggttc tggttttact gacacctatt tgaaggaata atttctgctc acacacagtt
     4681 ttgtaaccgc tcactattgt gttacagctc atcataagct actataacac atctttgcgg
     4741 ctttaagcta gtgtattcat ctaatttaca atcatgcgcg agtgtatctc tattcccggg
     4801 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     4861 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     4921 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     4981 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     5041 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     5101 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     5161 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     5221 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     5281 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     5341 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     5401 cctcgagcat cgatagacat gataagatac attgatgagt ttggacaaac cacaacaaga
     5461 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     5521 attataagct gcaataaaca agttaacaac aacaattcta gagggcctgc ttgccgaata
     5581 tcatggtgga aaatggccgc ttttc
//
