LOCUS       sequence-192713-        6271 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 6271)
  AUTHORS   Pennington HG, Youles M, Kamoun S
  TITLE     Golden-Gate compatible Magnaporthe oryzae protoplast transformation 
            vectors.
  JOURNAL   Figshare (2017)
REFERENCE   2  (bases 1 to 6271)
  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..6271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(423..434)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2517..2539
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2531..2548
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2532..2548
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2558..2577
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2558..2576
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2621..2648)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2664..2735)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2780..3457)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             3464..3475
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    3493..3509
                     /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(3517..3523,3524..3541,3542..3547))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3562..3583
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3724..3744)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3724..3742)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3763..3779)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3763..3779)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3787..3803
                     /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(3811..3817,3818..3835,3836..3841))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3856..3877
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3994..4011)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4165..4753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4245..4264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4924..5715,5716..5784))
                     /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     5547..5566
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acaaacaggc agagttacaa acgccatcac tacgccttgt cttttcgtct ctgttacgtg
       61 tcgccacccc catgcgagct acacgtctgc gcctgaagtc acaaggcttg caatttcatg
      121 acgatgaaca acactgacac ttctgttaca gagcacctcg aactaagccc agcgccagct
      181 tcaatgctcg ccgcgatccc cagcctctgg tcaaccctcg ctcaggtgag gcagacgaat
      241 cgtaagttgc gccaacaccg ctcctacacc gccaagaccc ataccgccgt cgcatttgga
      301 caagcacaag actgatttga ccgtggcata gattcattgg caagacggga ggagagattt
      361 tccacgagat gatgctgagg caaaacgtca agcatatctg taagcgtcta attctacaaa
      421 ttcctaccct cgatttcaac cacatattca accacatatt ctgacacatg agtcacagtc
      481 ggttaccctg gcggtgctat ccttcccgtg ttcgacgcga tctacaactc gaagcacatc
      541 gactttgttc tgcccaagca tgagcaaggc gccggccaca tggcagaggg ctatgctcgc
      601 gcttcaggca aacccggcgt tgttctcgtc acctccggcc ccggtgccac aaatgtcatc
      661 actcccatgg ccgacgctct tgccgacggt acacctctgg ttgtattctc aggacaggtt
      721 gttacctctg atattggaag cgacgccttc caggaggccg acgtcatagg catctcccgg
      781 tcttgcacca agtggaacgt catggttaag agcgctgacg agctcccgag gagaattaac
      841 gaggcctttg agattgccac cagtgggcga cctgggcctg tcttggtcga tctgcccaag
      901 gacgtcacgg ctagtgtgct gaggagggct atccccaccg aaacctcgat tccctctatt
      961 agcgcagcag cacgggctgt ccaagaggca ggccgaaagc agcttgagca ctccatcaaa
     1021 cgcgtagccg atctcgtcaa cattgccaag aagcccgtca tatatgccgg ccaaggtgtc
     1081 attttgtcgg aaggcggcgt tgaacttctc aaggcgcttg ccgacaaggc ctcgattcct
     1141 gtcaccacca ctctgcatgg tctgggagcc tttgacgagc tcgacgagaa ggcactgcac
     1201 atgcttggta tgcacggttc ggcttatgcc aacatgtcca tgcaagaggc cgatttgatc
     1261 attgcccttg gtggccgctt cgatgaccgt gtcactggca gcatccccaa atttgctcct
     1321 gccgccaagc tagctgctgc tgaaggacgc ggaggtattg tccacttcga gattatgccc
     1381 aagaacatca acaaggtcgt ccaagcaaca gaggccattg agggcgacgt tgcttcgaac
     1441 ttgaagctgt tgctccccaa gattgaacaa cgatccatga ccgatcgcaa ggagtggttc
     1501 gaccagatca aggagtggaa ggagaagtgg cctctgtcac attatgagag ggccgagcgt
     1561 agtggactca tcaagcctca gactctgatc gaggagctga gcaacctgac tgctgaccgc
     1621 aaggacatga cctacatcac aaccggtgtt ggccagcacc aaatgtggac agcacaacat
     1681 ttcaggtgga ggcacccacg gtccatgatc acctctggcg gtttgggaac catgggatat
     1741 ggtctgccgg cagcgattgg cgccaaggtt gctaggccag atgctttggt cattgacatc
     1801 gacggcgacg catcgttcaa catgactctg acagagcttt cgacggcggc acagttcaac
     1861 attggcgtca aggtcattgt cttgaacaac gaggagcagg gaatggtgac ccaatggcag
     1921 aacttgttct acgaggaccg ctactcacat acacaccagc gcaacccaga cttcatgaag
     1981 ctcgccgatg caatggacgt tcaacatcgc cgtgtttcga agcctgacga tgtcgttgat
     2041 gctctgacgt ggctgatcaa caccgacggc cccgctctgc ttgaggtgat gactgataag
     2101 aaggttcctg ttctgcccat ggtgcccgga ggtaacggcc tgcacgagtt catcacgttt
     2161 gatgccagta agtaatatat ccaatgcact cctgtggccc attttgagcc ttgggtatga
     2221 ttctagttca acttgctaac cagtgcatca actcgtatag gcaaggataa gcaacggaga
     2281 gagctgatgc gcgcgaggac gaatggcctg cacggttaag ttgataatgg gaattgatta
     2341 ttgcacggga attgcatgct ctcacgtcga gcccattaac gcttacaatt tccattcgcc
     2401 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     2461 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     2521 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
     2581 attgggtacc gggccccccc tcgagcgtga gacctataaa cgcagaaagg cccacccgaa
     2641 ggtgagccag tgtgactcta gtagagagcg ttcaccgaca aacaacagat aaaacgaaag
     2701 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggctctagt agcgatctac
     2761 actagcacta tcagcgttat taagcaccgg tggagtgacg accttcagca cgttcgtact
     2821 gttcaacgat ggtgtagtcc tcgttgtggg aggtgatgtc cagtttgatg tcggttttgt
     2881 aagcacccgg cagctgaacc ggttttttag ccatgtaggt ggttttaact tcagcgtcgt
     2941 agtgaccacc gtctttcagt ttcagacgca ttttgatttc acctttcaga gcaccgtcct
     3001 ccgggtacat acgttcggtg gaagcttccc aacccatggt ttttttctgc ataaccggac
     3061 cgtcggacgg gaagttggta ccacgcagtt taactttgta gatgaactca ccgtcttgca
     3121 gggaggagtc ctgggtaacg gtaacaacac caccgtcctc gaagttcata acacgttccc
     3181 atttgaaacc ttccgggaag gacagtttca ggtagtccgg gatgtcagcc gggtgtttaa
     3241 cgtaagcttt ggaaccgtac tggaactgcg gggacaggat gtcccaagcg aacggcagcg
     3301 gaccaccttt ggtaactttc agtttagcgg tctgggtacc ttcgtacgga cgaccttcac
     3361 cttcaccttc gatttcgaac tcgtgaccgt taacggaacc ttccatacga actttgaaac
     3421 gcatgaactc tttgataacg tcctcggagg aagccatcta gtatttctcc tctttctcta
     3481 gtatgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3541 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3601 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3661 ggggagaggc ggtttgcgta ttgatatcgg tctcactccg cggtggagct ccagcttttg
     3721 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     3781 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3841 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3901 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3961 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4021 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4081 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4141 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4201 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4261 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4321 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4381 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4441 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4501 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4561 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4621 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4681 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4741 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4801 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4861 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4921 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4981 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5041 ccccagtgct gcaatgatac cgcgtgaccc acgctcaccg gctccagatt tatcagcaat
     5101 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5161 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5221 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5281 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5341 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5401 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5461 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5521 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5581 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5641 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5701 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5761 gacacggaaa tgttgaatac tcatactctt cctttttcaa tgggctgcac ctcgacgtgc
     5821 caacgccaca gtgccccaca tctcccggct ggattcaccg ttgagcgggg attgtgtgga
     5881 gggacttggt cgaaaatttg tgaacctccg atggcggcaa acatgactcc acgattcaca
     5941 gcccaanggc ctggccctgc ttcaattgtc atcgtctgac aggtccagga aaaatagtta
     6001 gctggaggct cgttactttt ttttcccgct tcaacggcca agcaagcaag ctcatcgtct
     6061 ctcgttcttt tctccattat tgacgactga naacagattc gaaatgcttc gtactgttgg
     6121 ccgcaaagcc ctgaggggct catccaaggg atgttcacga accatctcga ctctcaagcc
     6181 cgccacggca actattgcca agcccggcag caggaccctt tcgacgccag cgacggcaac
     6241 agcaacgtaa gtcagaatca cgagcgagcg a
//
