LOCUS       sequence-213489-        6230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid expresses Venus fluorescent protein fused to endogenous
            Histone H2B (H2B) gene of Capsaspora (CAOG_01818). It can be used to
            transfect Capsaspora cells and visualize nucleus in vivo..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6230)
  AUTHORS   Parra-Acero H, Ros-Rocher N, Perez-Posada A, Kozyczkowska A, 
            Sanchez-Pons N, Nakata A, Suga H, Najle SR, Ruiz-Trillo I
  TITLE     Transfection of Capsaspora owczarzaki, a close unicellular relative 
            of animals.
  JOURNAL   Development. 2018 May 11. pii: dev.162107. doi: 10.1242/dev.162107.
  PUBMED    29752387
REFERENCE   2  (bases 1 to 6230)
  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: 
            "Development. 2018 May 11. pii: dev.162107. doi: 
            10.1242/dev.162107."
COMMENT     Sequence Label: pONSY-CoH2B:Venus
FEATURES             Location/Qualifiers
     source          1..6230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    81..102
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(117..122,123..140,141..147)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    155..171
                     /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     160..182
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     179..195
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     179..195
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(1529..1531,1532..1534,1535..2248)
                     /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
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1574..1595)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1835..1854)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2182..2203
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2637..2656)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2638..2656)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2663..2680)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2663..2679)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2672..2694)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2820..3248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2907..2926)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3117..3138
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3592..4386
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3646..3665)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4256..4275
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4404..4472,4473..5264)
                     /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(4622..4641)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5435..6023
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5924..5943
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6177..6194
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
       61 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
      121 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
      181 ggaaacagct atgaccatga ttacgccaag cttggtacca aatgcacagt tagcaacgac
      241 cgacaatcag caacctccaa actctcagtt tttttccccc aatcgttgtc gatttagctt
      301 ttgatgctgc attttgaaat agaacgacaa ccgaatttgg gagagagcac tcttttacag
      361 gtttgtgaaa agaaaaaaaa tcgaacgtga cttgcttcca tcttccttct cgaagccgat
      421 ctcggcccat ccccgcttgt cccctcctcc cctcctcgca atccgctaga cagtgtctct
      481 gcacactgga ttgcggtgcg cgggccatcc gatgcccctt tcctcggcct ggagatggac
      541 gtgaaagtgg aaaacgaaaa acgcgaaaca gaaacatgtg gttctttttt ttttcttttt
      601 ctttttcgcc ttcgggcttt tctctctcgc ttcctcttct ccgctctctt cttcgcgctc
      661 ttttgcctct tgcgtgagat gctcgtcttg caatcgtctg ccactgctgc gctgtcacga
      721 tcgatcgcat cggacaagcc gtttctgttt tgaaatttag aacgttctgg aaaacaacaa
      781 aaaattgcct ttttttcact tttttctttt ttcgtttttt tccctcttca acacttcctc
      841 caactgtcgt tctgcgtttg ttcttcgctc tcgtcgtgca cactcggatt aggatccatt
      901 catccccatc cccttgcgca ttgccaaaaa cgagcgtcga aacgcgaacg tcagcgtggg
      961 gtgttgacgc cgaaccccac gagtgttttt ttttaaaatg ctgtttcatc gttttcttcc
     1021 ttctccccac gtgcaatcct gatttaaata attgtcgctt tttgatttgg aaatgctcac
     1081 tcttccattt ttttattatt taacacacac agaacaacct tcacaggatc ccgggatgcc
     1141 gccgaaggtc gcaaagaagg gtgagaagcg tgccggcaag gccgctgcca aggctgcctc
     1201 tggcgccgac aaggacaaga agaagaagaa gcgccgcaag gaatcgtacg gcatctacat
     1261 ctacaaggtc ctccgccaag tccacgccga caccggcatc tccaaccgcg ccatgaacat
     1321 catgaactcg ttcgtcaacg acatcttcga gcgcattgct ggcgaggcgt cccgtctggc
     1381 ccacttcaac aaccgctcga ccatctcgtc tcgcgagatc cagactgccg tccgtctcct
     1441 cctccccggc gaacttgcca agcacgccgt ctctgagggc accaaggccg tcaccaagta
     1501 cacctccggc gccaagacta gtgatatcat ggtgagcaag ggcgaggagc tgttcaccgg
     1561 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     1621 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac
     1681 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg
     1741 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     1801 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     1861 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     1921 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     1981 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
     2041 catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2101 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga
     2161 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     2221 tctcggcatg gacgagctgt acaagtaaat tttgtgtttg ccaagacaca tttgtttatt
     2281 tcaatcattt ctaagaccaa aaagggattt catgattttt gttgcttgca ttgaagtttt
     2341 ttttggttgt catttttttt gcgttggtta tgagcgcaat atattccatt atggcaagaa
     2401 acacgacgag ttgtttttac agtggaggga tgagcaagga atggggaaat ggagaacaag
     2461 tggaatgaaa tggtttgact ttgtgcggcg aaagtgttgc cattttcaat catgctgcgc
     2521 ctggcaccta gatgcgtgtg attgctcatt gctagtgctg ttctcaccaa gccgaattct
     2581 gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc aattcgccct
     2641 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     2701 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     2761 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     2821 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2881 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2941 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3001 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3061 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3121 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3181 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3241 cgcgaatttt aacaaaattc agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa
     3301 gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca
     3361 agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag
     3421 ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct
     3481 ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga
     3541 tggcgcaggg gatcaagatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3601 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3661 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3721 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3781 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3841 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3901 tcatcccacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3961 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4021 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4081 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4141 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4201 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4261 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4321 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4381 ttctgaattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4441 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4501 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4561 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4621 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4681 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4741 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4801 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4861 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4921 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4981 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5041 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5101 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5161 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5221 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5281 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5341 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5401 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5461 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5521 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5581 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5641 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5701 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5761 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5821 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5881 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5941 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6001 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6061 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6121 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6181 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
//
