LOCUS       sequence-200865-        5676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry under tubulin 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 5676)
  AUTHORS   Inaki Ruiz Trillo'  Elena Casacuberta
  TITLE     Ruiz-Trillo Corallochytrium plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5676)
  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            "unpublished"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 Tub MCH (CTM)
FEATURES             Location/Qualifiers
     source          1..5676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    515..536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(551..556,557..574,575..581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    589..605
                     /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     594..616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     613..629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     613..629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1430..2140
                     /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(1575..1593)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1843..1862
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2037..2060
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2517..2536)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2518..2536)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2543..2560)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2543..2559)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2552..2574)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2700..3128
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2787..2806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2997..3018
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3472..4266
                     /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(3526..3545)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4136..4155
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4284..4352,4353..5144)
                     /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(4502..4521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5315..5676,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttatc ggtgggactg
      661 tgagaactgc tacctgcctg tgcagaggag agcacactac attcaccatt gaaaatatgg
      721 cccactgaac tgatcaagcc aagtgaagac tgcgcgcctg tgttctgatt cttcccatgg
      781 tacaagtact gaaactttgg aagcgccaga gaattcacag acgcacagaa gacagcgcat
      841 ggtggaggct gtgtgtgttc accgaggtgt cctctgcgga aggtgccgtt cattatgtat
      901 tcggagtaaa gcacaacctc ttcaatgctg ctcccttcct tcgcttgttt cttggatagt
      961 tatcgtctgt cagaatcttt cggtacattc agtatagagt tcgcgatcgg tagcaacagt
     1021 cggacggacc ctgtcaggac ggtgtattcg gtggcctact tttcttttgt tgcatagctg
     1081 ttcctgacgc gatgcgccaa gcgccgtcat attgtgctgt gtgcccctat gtggagcagg
     1141 catagagctc ttgccaaatg ttgggaagag ctgttggaga agcgacagag cagtgccaag
     1201 acgacgaacg aggtgtgcga gctgtagtgc tagtgcttcc taggttctgg ttttactgac
     1261 acctatttga aggaataatt tctgctcaca cacagttttg taaccgctca ctattgtgtt
     1321 acagctcatc ataagctact ataacacatc tttgcggctt taagctagtg tattcatcta
     1381 atttacaatc atgcgcgagt gtatctctat tcccgggact agtgatatca tggtgagcaa
     1441 gggcgaggag gacaacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     1501 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     1561 gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga
     1621 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     1681 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     1741 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     1801 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     1861 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     1921 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgccg aggtcaagac
     1981 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagct
     2041 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc gcgccgaggg
     2101 ccgccactcc accggcggca tggacgagct gtacaagtaa attttgtgtt tgccaagaca
     2161 catttgttta tttcaatcat ttctaagacc aaaaagggat ttcatgattt ttgttgcttg
     2221 cattgaagtt ttttttggtt gtcatttttt ttgcgttggt tatgagcgca atatattcca
     2281 ttatggcaag aaacacgacg agttgttttt acagtggagg gatgagcaag gaatggggaa
     2341 atggagaaca agtggaatga aatggtttga ctttgtgcgg cgaaagtgtt gccattttca
     2401 atcatgctgc gcctggcacc tagatgcgtg tgattgctca ttgctagtgc tgttctcacc
     2461 gcggtcatcc atcacactgg cggccgctcg agcatgcatc tagagggccc aattcgccct
     2521 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     2581 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     2641 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     2701 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2761 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2821 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2881 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2941 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3001 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3061 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3121 cgcgaatttt aacaaaattc agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa
     3181 gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca
     3241 agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag
     3301 ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct
     3361 ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga
     3421 tggcgcaggg gatcaagatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3481 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3541 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3601 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3661 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3721 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3781 tcatcccacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3841 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3901 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3961 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4021 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4081 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4141 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4201 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4261 ttctgaattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4321 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4381 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4441 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4501 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4561 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4621 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4681 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4741 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4801 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4861 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4921 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4981 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5041 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5101 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5161 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5221 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5281 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5341 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5401 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5461 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5521 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5581 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5641 gttcgtgcac acagcccagc ttggagcgaa cgacct
//
