LOCUS       sequence-204750-        4381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden-Gate compatible fungal transformation vector carrying BASTA 
            (Bialaphos/ Phosphinothricin) Resistance w 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 4381)
  AUTHORS   Pennington HG, Youles M, Kamoun S
  TITLE     Golden-Gate compatible Magnaporthe oryzae protoplast transformation 
            vectors.
  JOURNAL   Figshare (2017)
REFERENCE   2  (bases 1 to 4381)
  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: pCB1532B-RFP
FEATURES             Location/Qualifiers
     source          1..4381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     153..170
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    287..308
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(323..328,329..346,347..353)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    361..377
                     /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     385..401
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     385..401
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     420..440
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        422..440
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    584..605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(620..625,626..643,644..650)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    658..674
                     /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)."
     RBS             692..703
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             710..1387
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1432..1503
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1519..1546
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1590..1609)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1591..1609)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1619..1636)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1619..1635)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1628..1650)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        1801..2160
                     /label=trpC promoter
                     /note="promoter for Aspergillus nidulans trpC"
     CDS             2183..2734
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     CDS             join(2761..2829,2830..3621)
                     /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(2979..2998)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3792..4380
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4281..4300
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
       61 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
      121 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
      181 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
      241 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      301 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      361 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      421 caattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtgga gtgagaccga
      481 tatcaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
      541 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
      601 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
      661 tgagcggata acaatttcac acatactaga gaaagaggag aaatactaga tggcttcctc
      721 cgaggacgtt atcaaagagt tcatgcgttt caaagttcgt atggaaggtt ccgttaacgg
      781 tcacgagttc gaaatcgaag gtgaaggtga aggtcgtccg tacgaaggta cccagaccgc
      841 taaactgaaa gttaccaaag gtggtccgct gccgttcgct tgggacatcc tgtccccgca
      901 gttccagtac ggttccaaag cttacgttaa acacccggct gacatcccgg actacctgaa
      961 actgtccttc ccggaaggtt tcaaatggga acgtgttatg aacttcgagg acggtggtgt
     1021 tgttaccgtt acccaggact cctccctgca agacggtgag ttcatctaca aagttaaact
     1081 gcgtggtacc aacttcccgt ccgacggtcc ggttatgcag aaaaaaacca tgggttggga
     1141 agcttccacc gaacgtatgt acccggagga cggtgctctg aaaggtgaaa tcaaaatgcg
     1201 tctgaaactg aaagacggtg gtcactacga cgctgaagtt aaaaccacct acatggctaa
     1261 aaaaccggtt cagctgccgg gtgcttacaa aaccgacatc aaactggaca tcacctccca
     1321 caacgaggac tacaccatcg ttgaacagta cgaacgtgct gaaggtcgtc actccaccgg
     1381 tgcttaataa cgctgatagt gctagtgtag atcgctacta gagccaggca tcaaataaaa
     1441 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1501 ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta taggtctcac
     1561 gctcgagggg gggcccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     1621 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     1681 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     1741 cttcccaaca gttgcgcagc ctgaatggcg aatggaaatt gtaagcgtta atgggctcga
     1801 cagaagatga tattgaagga gcactttttg ggcttggctg gagctagtgg aggtcaacaa
     1861 tgaatgccta ttttggttta gtcgtccagg cggtgagcac aaaatttgtg tcgtttgaca
     1921 agatggttca tttaggcaac tggtcagatc agccccactt gtagcagtag cggcggcgct
     1981 cgaagtgtga ctcttattag cagacaggaa cgaggacatt attatcatct gctgcttggt
     2041 gcacgataac ttggtgcgtt tgtcaagcaa ggtaagtgaa cgacccggtc ataccttctt
     2101 aagttcgccc ttcctccctt tatttcagat tcaatctgac ttacctattc tacccaagca
     2161 acgcttcgat taggaagtaa ccatgagccc agaacgacgc ccggccgaca tccgccgtgc
     2221 caccgaggcg gacatgccgg cggtctgcac catcgtcaac cactacatcg agacaagcac
     2281 ggtcaacttc cgtaccgagc cgcaggaacc gcaggagtgg acggacgacc tcgtccgtct
     2341 gcgggagcgc tatccctggc tcgtcgccga ggtggacggc gaggtcgccg gcatcgccta
     2401 cgcgggtccc tggaaggcac gcaacgccta cgactggacg gccgaatcga ccgtgtacgt
     2461 ctccccccgc caccagcgga cgggactggg ctccacgctc tacacccacc tgctgaagtc
     2521 cctggaggca cagggcttca agagcgtggt cgctgtcatc gggctgccca acgacccgag
     2581 cgtgcgcatg cacgaggcgc tcggatatgc cccccgcggc atgctgcggg cggccggctt
     2641 caagcacggg aactggcatg acgtgggttt ctggcagctg gacttcagcc tgccggttcc
     2701 gccccgtccg gtcctgcccg tcaccgagat ttaggtcgag cccattgaaa aaggaagagt
     2761 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     2821 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2881 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     2941 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3001 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3061 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3121 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3181 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3241 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3301 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3361 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3421 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtca
     3481 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     3541 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     3601 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     3661 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3721 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3781 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3841 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3901 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3961 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4021 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4081 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4141 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4201 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4261 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4321 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4381 a
//
