LOCUS       sequence-213490-        5904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid expresses mCherry fluorescent protein fused to an 
            N-Myristoylation motif (NMM) from the endogenous Src2 gene 
            (CAOG_06360). It can be used to visualize the membrane and 
            filopodia..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5904)
  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 5904)
  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-CoNMM:mCherry
FEATURES             Location/Qualifiers
     source          1..5904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     509..528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     762..779
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    896..917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(932..937,938..955,956..962)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    970..986
                     /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     975..997
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     994..1010
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     994..1010
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2026..2736
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2171..2189)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2439..2458
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2633..2656
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3126..3145)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3127..3145)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3152..3169)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3152..3168)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3161..3183)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3309..3737
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3396..3415)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3606..3627
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             4081..4875
                     /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(4135..4154)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4745..4764
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4893..4961,4962..5753)
                     /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(5111..5130)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
       61 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      661 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      721 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      781 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      841 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      901 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      961 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     1021 ccaagcttgg taccaaatgc acagttagca acgaccgaca atcagcaacc tccaaactct
     1081 cagttttttt cccccaatcg ttgtcgattt agcttttgat gctgcatttt gaaatagaac
     1141 gacaaccgaa tttgggagag agcactcttt tacaggtttg tgaaaagaaa aaaaatcgaa
     1201 cgtgacttgc ttccatcttc cttctcgaag ccgatctcgg cccatccccg cttgtcccct
     1261 cctcccctcc tcgcaatccg ctagacagtg tctctgcaca ctggattgcg gtgcgcgggc
     1321 catccgatgc ccctttcctc ggcctggaga tggacgtgaa agtggaaaac gaaaaacgcg
     1381 aaacagaaac atgtggttct tttttttttc tttttctttt tcgccttcgg gcttttctct
     1441 ctcgcttcct cttctccgct ctcttcttcg cgctcttttg cctcttgcgt gagatgctcg
     1501 tcttgcaatc gtctgccact gctgcgctgt cacgatcgat cgcatcggac aagccgtttc
     1561 tgttttgaaa tttagaacgt tctggaaaac aacaaaaaat tgcctttttt tcactttttt
     1621 cttttttcgt ttttttccct cttcaacact tcctccaact gtcgttctgc gtttgttctt
     1681 cgctctcgtc gtgcacactc ggattaggat ccattcatcc ccatcccctt gcgcattgcc
     1741 aaaaacgagc gtcgaaacgc gaacgtcagc gtggggtgtt gacgccgaac cccacgagtg
     1801 ttttttttta aaatgctgtt tcatcgtttt cttccttctc cccacgtgca atcctgattt
     1861 aaataattgt cgctttttga tttggaaatg ctcactcttc cattttttta ttatttaaca
     1921 cacacagaac aaccttcaca ggatcccggg atgggctgct ccaactctaa accgcacgac
     1981 ccgtccgatt tcaaggtttc cccttctggc gttgcgtcca acagcatggt gagcaagggc
     2041 gaggaggaca acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     2101 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2161 acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
     2221 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     2281 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     2341 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     2401 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     2461 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     2521 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgccgaggt caagaccacc
     2581 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagctggac
     2641 atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgcgc cgagggccgc
     2701 cactccaccg gcggcatgga cgagctgtac aagtaaatct gaattttgtg tttgccaaga
     2761 cacatttgtt tatttcaatc atttctaaga ccaaaaaggg atttcatgat ttttgttgct
     2821 tgcattgaag ttttttttgg ttgtcatttt ttttgcgttg gttatgagcg caatatattc
     2881 cattatggca agaaacacga cgagttgttt ttacagtgga gggatgagca aggaatgggg
     2941 aaatggagaa caagtggaat gaaatggttt gactttgtgc ggcgaaagtg ttgccatttt
     3001 caatcatgct gcgcctggca cctagatgcg tgtgattgct cattgctagt gctgttctca
     3061 ccgcggaagc tttcaatcca tcacactggc ggccgctcga gcatgcatct agagggccca
     3121 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     3181 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     3241 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     3301 gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3361 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3421 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3481 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3541 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3601 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3661 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3721 aaaatttaac gcgaatttta acaaaattca gggcgcaagg gctgctaaag gaagcggaac
     3781 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct
     3841 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca
     3901 tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
     3961 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
     4021 aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatgagg atcgtttcgc
     4081 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4141 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4201 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4261 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4321 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4381 gatctcctgt catcccacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4441 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4501 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4561 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4621 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4681 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4741 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4801 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4861 gacgagttct tctgaattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4921 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4981 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5041 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5101 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5161 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5221 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5281 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5341 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5401 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5461 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5521 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5581 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5641 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5701 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5761 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5821 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5881 tccactgagc gtcagacccc gtag
//
