LOCUS       sequence-213485-        5849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid expresses Venus fluorescent protein under endogenous 
            EF1a promoter. It can be used to transfect ""Capsaspora owczarzaki""
            cells and visualize them by fluorescence microscopy..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5849)
  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 5849)
  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-Venus
FEATURES             Location/Qualifiers
     source          1..5849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(1173..1175,1176..1178,1179..1892)
                     /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(1218..1239)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1479..1498)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1826..1847
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2281..2300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2282..2300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2307..2324)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2307..2323)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2316..2338)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2464..2892
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2551..2570)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2761..2782
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3236..4030
                     /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(3290..3309)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3900..3919
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4048..4116,4117..4908)
                     /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(4266..4285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5079..5667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5568..5587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5821..5838
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgg
      241 taccaaatgc acagttagca acgaccgaca atcagcaacc tccaaactct cagttttttt
      301 cccccaatcg ttgtcgattt agcttttgat gctgcatttt gaaatagaac gacaaccgaa
      361 tttgggagag agcactcttt tacaggtttg tgaaaagaaa aaaaatcgaa cgtgacttgc
      421 ttccatcttc cttctcgaag ccgatctcgg cccatccccg cttgtcccct cctcccctcc
      481 tcgcaatccg ctagacagtg tctctgcaca ctggattgcg gtgcgcgggc catccgatgc
      541 ccctttcctc ggcctggaga tggacgtgaa agtggaaaac gaaaaacgcg aaacagaaac
      601 atgtggttct tttttttttc tttttctttt tcgccttcgg gcttttctct ctcgcttcct
      661 cttctccgct ctcttcttcg cgctcttttg cctcttgcgt gagatgctcg tcttgcaatc
      721 gtctgccact gctgcgctgt cacgatcgat cgcatcggac aagccgtttc tgttttgaaa
      781 tttagaacgt tctggaaaac aacaaaaaat tgcctttttt tcactttttt cttttttcgt
      841 ttttttccct cttcaacact tcctccaact gtcgttctgc gtttgttctt cgctctcgtc
      901 gtgcacactc ggattaggat ccattcatcc ccatcccctt gcgcattgcc aaaaacgagc
      961 gtcgaaacgc gaacgtcagc gtggggtgtt gacgccgaac cccacgagtg ttttttttta
     1021 aaatgctgtt tcatcgtttt cttccttctc cccacgtgca atcctgattt aaataattgt
     1081 cgctttttga tttggaaatg ctcactcttc cattttttta ttatttaaca cacacagaac
     1141 aaccttcaca ggatcccggg actagtgata tcatggtgag caagggcgag gagctgttca
     1201 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1261 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca
     1321 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcctgc
     1381 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1441 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1501 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1561 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1621 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     1681 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1741 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca
     1801 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1861 tcactctcgg catggacgag ctgtacaagt aaattttgtg tttgccaaga cacatttgtt
     1921 tatttcaatc atttctaaga ccaaaaaggg atttcatgat ttttgttgct tgcattgaag
     1981 ttttttttgg ttgtcatttt ttttgcgttg gttatgagcg caatatattc cattatggca
     2041 agaaacacga cgagttgttt ttacagtgga gggatgagca aggaatgggg aaatggagaa
     2101 caagtggaat gaaatggttt gactttgtgc ggcgaaagtg ttgccatttt caatcatgct
     2161 gcgcctggca cctagatgcg tgtgattgct cattgctagt gctgttctca ccaagccgaa
     2221 ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagagg gcccaattcg
     2281 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     2341 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2401 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2461 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2521 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2581 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2641 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2701 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2761 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2821 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2881 ttaacgcgaa ttttaacaaa attcagggcg caagggctgc taaaggaagc ggaacacgta
     2941 gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg
     3001 gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg
     3061 atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc
     3121 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
     3181 ctgatggcgc aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3241 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3301 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3361 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3421 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3481 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3541 cctgtcatcc caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3601 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3661 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3721 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3781 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3841 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3901 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3961 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4021 gttcttctga attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4081 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4141 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4201 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4261 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4321 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4381 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4441 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4501 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4561 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4621 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4681 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4741 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4801 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4861 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4921 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4981 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5041 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5101 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5161 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5221 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5281 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5341 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5401 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5461 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5521 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5581 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5641 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5701 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5761 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5821 agcgagtcag tgagcgagga agcggaaga
//
