LOCUS       sequence-200869-        5127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry under endogenous actin promoter. It can be used to
            transfect Corallochytrium limacisporum (Corallochytrea) cells and 
            visualize them by fluorescent microscope. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5127)
  AUTHORS   Inaki Ruiz Trillo'  Elena Casacuberta
  TITLE     Ruiz-Trillo Corallochytrium plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5127)
  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: 
            "unpublished"
COMMENT     Sequence Label: Clim Act MCH (CAM)
FEATURES             Location/Qualifiers
     source          1..5127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     146..165
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     340..363
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     2101..2120
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2249..2317,2318..3109)
                     /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(2467..2486)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3280..3868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3769..3788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4022..4039
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4156..4177
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4192..4197,4198..4215,4216..4222)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4230..4246
                     /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     4235..4257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4254..4270
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4254..4270
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(4860..5127,1..440)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGAHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5005..5023)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
ORIGIN
        1 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
       61 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
      121 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
      181 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
      241 tgaagctgaa ggacggcgcc cactacgacg ccgaggtcaa gaccacctac aaggccaaga
      301 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gctggacatc acctcccaca
      361 acgaggacta caccatcgtg gaacagtacg agcgcgccga gggccgccac tccaccggcg
      421 gcatggacga gctgtacaag gcggccgcca gtgtgatgga tatctgcaga attcggcttg
      481 tcgacatggg catggagaca gtagagcaac tacgcaagac aactattgtc tgcgacaatg
      541 gtaccggttt cgttaagtgc ggctacgccg acgacaagta ccctcgggac atgttcccga
      601 gtttggtcgg acggccattc cttcgcgtag actccactgt gggaagtaca aaactggatg
      661 atatcatggt cggacaggag gctagtgaca atcgggcaaa ccttgacctc cgctacccca
      721 tggagaatgg tatcattaaa cactgggatg acatggagcg gatctgggac cacactttct
      781 acgatcgtat gaagagcaac ccgaaggaat gcaaggttct tctcaccgag gccccacgga
      841 atccaatcag caaccgtaag cagatgattg acattatgtt tgagaagtac ggattccaaa
      901 gcgtgtacgt gggtacccag gccgtgctag tgctgtacgc ccagggtctg acaaccggtg
      961 tggtcgtgga ctctggcgat ggtgttactc atattattcc tgtgtatgag ggtcatgtcc
     1021 tgaaggatag catgcgccgt cttgatgtgg caggccgcga cattactcgt tacttgatta
     1081 agctactgct cctgcgggga tacgctttca accagagcgc tgactacgag acggtgcgtc
     1141 agatcaagga gaaactctgc tacgtagcct atgacactaa gcaggaacaa aaactcgcat
     1201 tagaaaccac cactctaatg gagaagtacg agctaccaga cggtcgcgtg atttcgttgg
     1261 gcagtgagcg cttcgaggct gccgaggccc tcttccaacc ccacctgatc aacgtcgaac
     1321 aggccggcat ggccgagcag ttgttcaact ccattcagag tacagatatt gacacacggt
     1381 cgacctttta caagaatgtt gtgctcagtg gtggatccac aatgtatccg ggcaccccaa
     1441 gtcgcattga gcgtgagatg aagcagctat atgccgagcg taccctgaag ggtgaccttt
     1501 cccggttgtc caaatttaag attaaggtca tggacccccc gaaccgtaag cacatggtgt
     1561 ttaatggcgg tgctgtcctt gctgagatta tgagggaccg gcctgacttt tggattagca
     1621 aagcggagtg ggatgaagat cctacacgtg ctatgcagaa gttgcagctc gagtaaacaa
     1681 atgttttgtt ggaagatttg atttgtcttt tttggagtgc ttagttagtg cactcctaga
     1741 tgagcaaaca ttaccacagt gtatcgcagc ataaaataaa attataaaat gtctttgtag
     1801 atcaaaagca ttacaaacaa atatttaaac tttgatcact tttttcaaat tcgtcaatat
     1861 gactaattaa catctgggtt atagatgaca agcatgaatg tttcatttat tttaaaaaaa
     1921 aaataaataa gtattttttc atttgcatca ccatcaaaaa tatttccatt gaaaagtcat
     1981 cgactggcta tttaaatttc tagagggcct gcttgccgaa tatcatggtg gaaaatggcc
     2041 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     2101 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     2161 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     2221 agttcttctg aattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2281 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2341 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2401 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2461 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     2521 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2581 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2641 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2701 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2761 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2821 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2881 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2941 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3001 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3061 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3121 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3181 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3241 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3301 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3361 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3421 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3481 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3541 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3601 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3661 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3721 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3781 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3841 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3901 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3961 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4021 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     4081 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     4141 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     4201 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     4261 cagctatgac catgattacg ccaagcttga aatgctgggc gccttgaaac tgaacctgtt
     4321 cggatccggg ttgcgcgctt ccattgtttt ggtttccgtg tctgcaaggc gggtctggac
     4381 gtgacgcttt gggggagagg gagagcagag atacaattct agccacgaat ttcagccaat
     4441 agtcacagag gggagttgcg atcctgcgaa ttggcttagg tctgtgaggt ctttcacggt
     4501 ttcgaaagca gcgcagaggg gagaccgact accaaaagcc ataacactcc accgagatat
     4561 ccgcactctt cgctagatga gcttgcttct cctcttgtca caaatcagcc caatgatagt
     4621 cactgttcaa caacggtgca gtgtaaatat tcgttgtgcg tccttgatga gcagaaggta
     4681 cgagaagcag gaaatcattt cataatggaa aaccattcac tgtctttctg tattgcgaaa
     4741 agagtgatca taggtaatcg caacggcggg tctttagaac agtcacatac cagatcatgg
     4801 gcatggagac agtagagcaa ctacgcaaga caactattgt ccccgggact agtgatatca
     4861 tggtgagcaa gggcgaggag gacaacatgg ccatcatcaa ggagttcatg cgcttcaagg
     4921 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     4981 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct
     5041 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     5101 ccgccgacat ccccgactac ttgaagc
//
