LOCUS       sequence-197099-        6274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden-Gate compatible fungal transformation vector carrying 
            Sulfonylurea Resistance (Chlorimuron Ethyl) with RFP screening 
            cassette (RFP knocked out by GG reaction, allowing for red-white 
            screening).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6274)
  AUTHORS   Pennington HG, Youles M, Kamoun S
  TITLE     Golden-Gate compatible Magnaporthe oryzae protoplast transformation 
            vectors.
  JOURNAL   Figshare (2017)
REFERENCE   2  (bases 1 to 6274)
  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: "Figshare 
            (2017)"
COMMENT     Sequence Label: pCB1532S-RFP
FEATURES             Location/Qualifiers
     source          1..6274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2021..2043
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2035..2052
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2036..2052
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2062..2081
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2062..2080
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2125..2152)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2168..2239)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2284..2961)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2968..2979
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2997..3013
                     /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)."
     promoter        complement(join(3021..3027,3028..3045,3046..3051))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3066..3087
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3231..3251)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3231..3249)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3270..3286)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3270..3286)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3294..3310
                     /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)."
     promoter        complement(join(3318..3324,3325..3342,3343..3348))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3363..3384
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3501..3518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3672..4260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3752..3771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4431..5222,5223..5291))
                     /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     5054..5073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(6201..6212)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 ctatccttcc cgtgttcgac gcgatctaca actcgaagca catcgacttt gttctgccca
       61 agcatgagca aggcgccggc cacatggcag agggctatgc tcgcgcttca ggcaaacccg
      121 gcgttgttct cgtcacctcc ggccccggtg ccacaaatgt catcactccc atggccgacg
      181 ctcttgccga cggtacacct ctggttgtat tctcaggaca ggttgttacc tctgatattg
      241 gaagcgacgc cttccaggag gccgacgtca taggcatctc ccggtcttgc accaagtgga
      301 acgtcatggt taagagcgct gacgagctcc cgaggagaat taacgaggcc tttgagattg
      361 ccaccagtgg gcgacctggg cctgtcttgg tcgatctgcc caaggacgtc acggctagtg
      421 tgctgaggag ggctatcccc accgaaacct cgattccctc tattagcgca gcagcacggg
      481 ctgtccaaga ggcaggccga aagcagcttg agcactccat caaacgcgta gccgatctcg
      541 tcaacattgc caagaagccc gtcatatatg ccggccaagg tgtcattttg tcggaaggcg
      601 gcgttgaact tctcaaggcg cttgccgaca aggcctcgat tcctgtcacc accactctgc
      661 atggtctggg agcctttgac gagctcgacg agaaggcact gcacatgctt ggtatgcacg
      721 gttcggctta tgccaacatg tccatgcaag aggccgattt gatcattgcc cttggtggcc
      781 gcttcgatga ccgtgtcact ggcagcatcc ccaaatttgc tcctgccgcc aagctagctg
      841 ctgctgaagg acgcggaggt attgtccact tcgagattat gcccaagaac atcaacaagg
      901 tcgtccaagc aacagaggcc attgagggcg acgttgcttc gaacttgaag ctgttgctcc
      961 ccaagattga acaacgatcc atgaccgatc gcaaggagtg gttcgaccag atcaaggagt
     1021 ggaaggagaa gtggcctctg tcacattatg agagggccga gcgtagtgga ctcatcaagc
     1081 ctcagactct gatcgaggag ctgagcaacc tgactgctga ccgcaaggac atgacctaca
     1141 tcacaaccgg tgttggccag caccaaatgt ggacagcaca acatttcagg tggaggcacc
     1201 cacggtccat gatcacctct ggcggtttgg gaaccatggg atatggtctg ccggcagcga
     1261 ttggcgccaa ggttgctagg ccagatgctt tggtcattga catcgacggc gacgcatcgt
     1321 tcaacatgac tctgacagag ctttcgacgg cggcacagtt cwacattggc gtcaaggtca
     1381 ttgtcttgaa caacgaggag cagggaatgg tgacccaatg gcagaacttg ttctacgagg
     1441 accgctactc acatacacac cagcgcaacc cagacttcat gaagctcgcc gatgcaatgg
     1501 acgttcaaca tcgccgtgtt tcgaagcctg acgatgtcgt tgatgctctg acgtggctga
     1561 tcaacaccga cggccccgct ctgcttgagg tgatgactga taagaaggtt cctgttctgc
     1621 ccatggtgcc cggaggtaac ggcctgcacg agttcatcac gtttgatgcc agtaagtaat
     1681 atatccaatg cactcctgtg gcccattttg agccttgggt atgattctag ttcaacttgc
     1741 taaccagtgc atcaactcgt ataggcaagg ataagcaacg gagagagctg atgcgcgcga
     1801 ggacgaatgg cctgcacggt taagttgata atgggaattg attattgcac gggaattgca
     1861 tgctctcacg tcgagcccat taacgcttac aatttccatt cgccattcag gctgcgcaac
     1921 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     1981 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     2041 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc
     2101 cccctcgagc gtgagaccta taaacgcaga aaggcccacc cgaaggtgag ccagtgtgac
     2161 tctagtagag agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact
     2221 gagcctttcg ttttatttga tgcctggctc tagtagcgat ctacactagc actatcagcg
     2281 ttattaagca ccggtggagt gacgaccttc agcacgttcg tactgttcaa cgatggtgta
     2341 gtcctcgttg tgggaggtga tgtccagttt gatgtcggtt ttgtaagcac ccggcagctg
     2401 aaccggtttt ttagccatgt aggtggtttt aacttcagcg tcgtagtgac caccgtcttt
     2461 cagtttcaga cgcattttga tttcaccttt cagagcaccg tcctccgggt acatacgttc
     2521 ggtggaagct tcccaaccca tggttttttt ctgcataacc ggaccgtcgg acgggaagtt
     2581 ggtaccacgc agtttaactt tgtagatgaa ctcaccgtct tgcagggagg agtcctgggt
     2641 aacggtaaca acaccaccgt cctcgaagtt cataacacgt tcccatttga aaccttccgg
     2701 gaaggacagt ttcaggtagt ccgggatgtc agccgggtgt ttaacgtaag ctttggaacc
     2761 gtactggaac tgcggggaca ggatgtccca agcgaacggc agcggaccac ctttggtaac
     2821 tttcagttta gcggtctggg taccttcgta cggacgacct tcaccttcac cttcgatttc
     2881 gaactcgtga ccgttaacgg aaccttccat acgaactttg aaacgcatga actctttgat
     2941 aacgtcctcg gaggaagcca tctagtattt ctcctctttc tctagtatgt gtgaaattgt
     3001 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3061 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3121 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3181 cgtattgata tcggtctcac tccaccgcgg tggagctcca gcttttgttc cctttagtga
     3241 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3301 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3361 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3481 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3541 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3601 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3661 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3721 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3781 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3841 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3901 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3961 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4021 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4081 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4141 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4201 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4261 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4321 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4381 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4441 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4501 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4561 atgataccgc gtgacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4621 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4681 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4741 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4801 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4861 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4921 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4981 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5041 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5101 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5161 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5221 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5281 tgaatactca tactcttcct ttttcaatgg gctgcacctc gacgtgccaa cgccacagtg
     5341 ccccacatct cccggctgga ttcaccgttg agcggggatt gtgtggaggg acttggtcga
     5401 aaatttgtga acctccgatg gcggcaaaca tgactccacg attcacagcc caaaggcctg
     5461 gccctgcttc aattgtcatc gtctgacagg tccaggaaaa atagttagct ggaggctcgt
     5521 tacttttttt tcccgcttca acggccaagc aagcaagctc atcgtctctc gttcttttct
     5581 ccattattga cgactgagaa cagattcgaa atgcttcgta ctgttggccg caaagccctg
     5641 aggggctcat ccaagggatg ttcacgaacc atctcgactc tcaagcccgc cacggcaact
     5701 attgccaagc ccggcagcag gaccctttcg acgccagcga cggcaacagc aacgtaagtc
     5761 agaatcacga gcgagcgaac aaacaggcag agttacaaac gccatcacta cgccttgtct
     5821 tttcgtctct gttacgtgtc gccaccccca tgcgagctac acgtctgcgc ctgaagtcac
     5881 aaggcttgca atttcatgac gatgaacaac actgacactt ctgttacaga gcacctcgaa
     5941 ctaagcccag cgccagcttc aatgctcgcc gcgatcccca gcctctggtc aaccctcgct
     6001 caggtgaggc agacgaatcg taagttgcgc caacaccgct cctacaccgc caagacccat
     6061 accgccgtcg catttggaca agcacaagac tgatttgacc gtggcataga ttcattggca
     6121 agacgggagg agagattttc cacgagatga tgctgaggca aaacgtcaag catatctgta
     6181 agcgtctaat tctacaaatt cctaccctcg atttcaacca catattcaac cacatattct
     6241 gacacatgag tcacagtcgg ttaccctggc ggtg
//
