LOCUS       sequence-192716-        4835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden-Gate compatible fungal transformation vector carrying 
            Hygromycin Resistance 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 4835)
  AUTHORS   Pennington HG, Youles M, Kamoun S
  TITLE     Golden-Gate compatible Magnaporthe oryzae protoplast transformation 
            vectors.
  JOURNAL   Figshare (2017)
REFERENCE   2  (bases 1 to 4835)
  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: pCB1532H-RFP
FEATURES             Location/Qualifiers
     source          1..4835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     146..168
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     160..177
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     161..177
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     187..206
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        187..205
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(250..277)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(293..364)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(409..1086)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             1093..1104
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    1122..1138
                     /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(1146..1152,1153..1170,1171..1176))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1191..1212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1353..1373)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1353..1371)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1392..1408)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1392..1408)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1416..1432
                     /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(1440..1446,1447..1464,1465..1470))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1485..1506
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1623..1640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1794..2382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1874..1893)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2553..3344,3345..3413))
                     /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     3176..3195
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3806..4831
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEXSRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
ORIGIN
        1 cccattaacg cttacaattt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
       61 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      121 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
      181 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgagcgtgag
      241 acctataaac gcagaaaggc ccacccgaag gtgagccagt gtgactctag tagagagcgt
      301 tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta
      361 tttgatgcct ggctctagta gcgatctaca ctagcactat cagcgttatt aagcaccggt
      421 ggagtgacga ccttcagcac gttcgtactg ttcaacgatg gtgtagtcct cgttgtggga
      481 ggtgatgtcc agtttgatgt cggttttgta agcacccggc agctgaaccg gttttttagc
      541 catgtaggtg gttttaactt cagcgtcgta gtgaccaccg tctttcagtt tcagacgcat
      601 tttgatttca cctttcagag caccgtcctc cgggtacata cgttcggtgg aagcttccca
      661 acccatggtt tttttctgca taaccggacc gtcggacggg aagttggtac cacgcagttt
      721 aactttgtag atgaactcac cgtcttgcag ggaggagtcc tgggtaacgg taacaacacc
      781 accgtcctcg aagttcataa cacgttccca tttgaaacct tccgggaagg acagtttcag
      841 gtagtccggg atgtcagccg ggtgtttaac gtaagctttg gaaccgtact ggaactgcgg
      901 ggacaggatg tcccaagcga acggcagcgg accacctttg gtaactttca gtttagcggt
      961 ctgggtacct tcgtacggac gaccttcacc ttcaccttcg atttcgaact cgtgaccgtt
     1021 aacggaacct tccatacgaa ctttgaaacg catgaactct ttgataacgt cctcggagga
     1081 agccatctag tatttctcct ctttctctag tatgtgtgaa attgttatcc gctcacaatt
     1141 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1201 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1261 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgatatcggt
     1321 ctcactccgc ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     1381 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1441 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1501 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1561 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1621 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1681 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1741 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1801 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1861 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1921 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1981 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2041 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2101 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2161 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2221 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2281 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2341 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2401 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2461 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2521 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2581 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2641 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgtgaccca
     2701 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2761 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2821 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2881 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     2941 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3001 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3061 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3121 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3181 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3241 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3301 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3361 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     3421 ctttttcaat gggtgatcga cgttaactga tattgaagga gcattttttg ggcttggctg
     3481 gagctagtgg aggtcaacaa tgaatgccta ttttggttta gtcgtccagg cggtgagcac
     3541 aaaatttgtg tcgtttgaca agatggttca tttaggcaac tggtcagatc agccccactt
     3601 gtagcagtag cggcggcgct cgaagtgtga ctcttattag cagacaggaa cgaggacatt
     3661 attatcatct gctgcttggt gcacgataac ttggtgcgtt tgtcaagcaa ggtaagtgga
     3721 cgacccggtc ataccttctt aagttcgccc ttcctccctt tatttcagat tcaatctgac
     3781 ttacctattc tacccaagca tccaaatgaa aaagcctgaa ctcaccgcga cgtctgtcga
     3841 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     3901 anaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     3961 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct
     4021 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc
     4081 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     4141 ccagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg
     4201 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     4261 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     4321 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     4381 gcacctcgtg catgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     4441 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     4501 cctcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     4561 gcatccggag cttgcaggat cgccgcgcct ccgggcgtat atgctccgca ttggtcttga
     4621 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     4681 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     4741 aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     4801 ccccagcact cgtccgaggg caaaggaata gcgag
//
