LOCUS       sequence-200864-        4307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry under elongation factor 1 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 4307)
  AUTHORS   Inaki Ruiz Trillo'  Elena Casacuberta
  TITLE     Ruiz-Trillo Corallochytrium plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 4307)
  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            "unpublished"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 EF1 MCH (CEM)
FEATURES             Location/Qualifiers
     source          1..4307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(13..18,19..36,37..43)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    51..67
                     /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     56..78
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     75..91
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     75..91
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1054..1764
                     /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(1199..1217)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1467..1486
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1661..1684
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     2229..2248
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2377..2445,2446..3237)
                     /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(2595..2614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3408..3996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3897..3916
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4150..4167
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4284..4305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
       61 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg gtaccgagct
      121 cggatccact agtaacggcc gccagtgtgc tggaattcgg cttctgggct tggaccatgt
      181 aatcgcatac gatgttactt gcctctccaa ctaatgctcc ccttgcaatg ttgttccttt
      241 tatgttggtt tctgtcgcat gtgcattgcc acatgcgacc ggtttcagat tctgtccaaa
      301 caagaaacca gcgaagcaac cccatcgctg ttctcccttc aagcaagagt gtgttgtttg
      361 gtggacggac atggaggtct gtttgtgaca agtccgcttt gctgcatcat tacccctgcc
      421 tgatctgtgt ctgtgcgttc gtatgtgtca gagctgcgac tgactcgttc ttccttcaga
      481 cccgggcttg gctatcctca tccacaacca tggcgagaga tcgagtactg acagacggag
      541 tatggatatg ggccgaatct ctccagggga tcttcaagaa gagtccgctg aaggcaatgt
      601 ggctgcccgg tgtgcccgtg tggtcgaccc cgcctcaata gccaatgtgg gtgatttgga
      661 gcaaccgtgt ggtcgcccct cttgcgccgt tgcattgttt tccctttgga acggctccgg
      721 aacaatggaa cattgggcac cactcatagt gtttctcaca cacacacaca cgctagcaaa
      781 cacacacaca tacacagaga ccaacacata tcgcagaagt tttcttttgc atacgatgtt
      841 cttggcatac aaacttcatt tcagtccctt tgaattttgg gcgcagtgat ggtatttgag
      901 accaccgaca tgttttgtaa acattcagta acacactgga aagtactgct gatctctagt
      961 tagttcaaca aacaaacagc aaacatgggt aaggaaaagc tccatcttaa cattgtcgtc
     1021 attggtcatg tcaagccgaa ttctgcagat atcatggtga gcaagggcga ggaggacaac
     1081 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     1141 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1201 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtcccctcag
     1261 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     1321 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     1381 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     1441 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     1501 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     1561 ctgaagctga aggacggcgg ccactacgac gccgaggtca agaccaccta caaggccaag
     1621 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agctggacat cacctcccac
     1681 aacgaggact acaccatcgt ggaacagtac gagcgcgccg agggccgcca ctccaccggc
     1741 ggcatggacg agctgtacaa gtaaattttg tgtttgccaa gacacatttg tttatttcaa
     1801 tcatttctaa gaccaaaaag ggatttcatg atttttgttg cttgcattga agtttttttt
     1861 ggttgtcatt ttttttgcgt tggttatgag cgcaatatat tccattatgg caagaaacac
     1921 gacgagttgt ttttacagtg gagggatgag caaggaatgg ggaaatggag aacaagtgga
     1981 atgaaatggt ttgactttgt gcggcgaaag tgttgccatt ttcaatcatg ctgcgcctgg
     2041 cacctagatg cgtgtgattg ctcattgcta gtgctgttct caccgcggtc atccatcaca
     2101 ctggcggccg ctcgagcatg catctagagg cgatgcctgc ttgccgaata tcatggtgga
     2161 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     2221 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     2281 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     2341 tcttgacgag ttcttctgaa ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2401 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2461 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2521 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2581 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2641 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2701 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2761 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2821 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2881 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2941 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3001 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3061 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3121 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3181 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3241 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3301 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3361 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3421 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3481 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3541 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3601 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3661 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3721 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3781 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3841 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3901 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3961 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4021 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4081 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4141 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     4201 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     4261 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcatta
//
