LOCUS       sequence-232217-        8098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Marker fusion of Bar:mCherry (without start codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8098)
  AUTHORS   Yadav V, Yang F, Reza MH, Liu S, Valent B, Sanyal K, Naqvi NI
  TITLE     Cellular Dynamics and Genomic Identity of Centromeres in Cereal 
            Blast Fungus.
  JOURNAL   MBio. 2019 Jul 30;10(4). pii: mBio.01581-19. doi: 
            10.1128/mBio.01581-19.
  PUBMED    31363034
REFERENCE   2  (bases 1 to 8098)
  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: "MBio."; 
            date: "2019-07-30"; volume: "10(4). pii: mBio.01581-19. doi"; pages:
            " 10.1128/mBio.01581-19"
COMMENT     Sequence Label: pFGL1269R
FEATURES             Location/Qualifiers
     source          1..8098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             24..569
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="SPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQEW
                     TDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGST
                     LYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGFW
                     QLDFSLPVPPRPVLPVTEI"
     CDS             570..1277
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     980..999
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1279..1295)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1279..1295)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1323..1340)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1323..1339)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1332..1354)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    1542..1566
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2867..3496
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3925..4998
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      5064..5258
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5494..5516)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5602..5742
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5657..5676
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5757..5774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5928..6516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6008..6027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6603..7397)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    7822..7846
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    7924..8098
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 ctcgagaatt cgagctcggt accagcccag aacgacgccc ggccgacatc cgccgtgcca
       61 ccgaggcgga catgccggcg gtctgcacca tcgtcaacca ctacatcgag acaagcacgg
      121 tcaacttccg taccgagccg caggaaccgc aggagtggac ggacgacctc gtccgtctgc
      181 gggagcgcta tccctggctc gtcgccgagg tggacggcga ggtcgccggc atcgcctacg
      241 cgggtccctg gaaggcacgc aacgcctacg actggacggc cgaatcgacc gtgtacgtct
      301 ccccccgcca ccagcggacg ggactgggct ccacgctcta cacccacctg ctgaagtccc
      361 tggaggcaca gggcttcaag agcgtggtcg ctgtcatcgg gctgcccaac gacccgagcg
      421 tgcgcatgca cgaggcgctc ggatatgccc cccgcggcat gctgcgggcg gccggcttca
      481 agcacgggaa ctggcatgac gtgggtttct ggcagctgga cttcagcctg ccggttccgc
      541 cccgtccggt cctgcccgtc accgagattg tgagcaaggg cgaggaggat aacatggcca
      601 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
      661 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
      721 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
      781 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
      841 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
      901 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
      961 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     1021 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     1081 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     1141 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     1201 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     1261 acgagctgta caagtaagga tccactagtt ctagagtcga cctgcaggca tgcaagcttg
     1321 gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     1381 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     1441 cgcccttccc aacagttgcg cagcctgaat ggcgaatgct agagcagctt gagcttggat
     1501 cagattgtcg tttcccgcct tcagtttaaa ctatcagtgt ttgacaggat atattggcgg
     1561 gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa
     1621 ggtttatccg ttggtccatt tgtatgtcca tgccaaccac agggttcccc tcgggatcaa
     1681 agtactttga tccaacccct ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag
     1741 ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg
     1801 cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac
     1861 tagaaccgga gacattacgc catgaacaag agcgccgccg ctggcctgct gggctatgcc
     1921 cgcgtcagca ccgacgacca ggacttgacc aaccaacggg ccgaactgca cgcggccggc
     1981 tgcaccaagc tgttttccga gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc
     2041 aggatgcttg accacctacg ccctggcgac gttgtgacag tgaccaggct agaccgcctg
     2101 gcccgcagca cccgcgacct actggacatt gccgagcgca tccaggaggc cggcgcgggc
     2161 ctgcgtagcc tggcagagcc gtgggccgac accaccacgc cggccggccg catggtgttg
     2221 accgtgttcg ccggcattgc cgagttcgag cgttccctaa tcatcgaccg cacccggagc
     2281 gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac cctcaccccg
     2341 gcacagatcg cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg
     2401 gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa
     2461 gtgacgccca ccgaggccag gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc
     2521 gacgccctgg cggccgccga gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg
     2581 acggccagga cgaaccgttt ttcattaccg aagagatcga ggcggagatg atcgcggccg
     2641 ggtacgtgtt cgagccgccc gcgcacgtct caaccgtgcg gctgcatgaa atcctggccg
     2701 gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa gaaaccgagc
     2761 gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg
     2821 cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga aggttatcgc
     2881 tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc
     2941 cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg gcagtgcccg
     3001 cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg accgcccgac
     3061 gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc
     3121 ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc tgattccggt
     3181 gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg ttaagcagcg
     3241 cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg
     3301 cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc ccattcttga
     3361 gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca caaccgttct
     3421 tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg ctgaaattaa
     3481 atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta
     3541 agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag cctggcagac
     3601 acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac caagctgaag
     3661 atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata catcgcgcag
     3721 ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg ctaaaggagg
     3781 cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg
     3841 aacgggcggt tggccaggcg taagcggctg ggttgcctgc cggccctgca atggcactgg
     3901 aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc
     3961 ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg
     4021 caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc tgatcgaatc
     4081 cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag
     4141 ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac ccgcgatagt
     4201 cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag
     4261 gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc ggccggcatg
     4321 gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac cgaatccatg
     4381 aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc acacgttgcg
     4441 gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga cctggtagaa
     4501 acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac
     4561 ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa gatcgtaaag
     4621 agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat gtaccgcgag
     4681 atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt tttgatcgat
     4741 cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc
     4801 agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt caagaagttc
     4861 tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga tttgaaggag
     4921 gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat cgagggcgaa
     4981 gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct agcaggggaa
     5041 aaaggtcgaa aaggtctctt tcctgtggat agcacgtaca ttgggaaccc aaagccgtac
     5101 attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa ccggtcacac
     5161 atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa
     5221 cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc
     5281 gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg
     5341 cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc aggcaatcta
     5401 ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc
     5461 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5521 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5581 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     5641 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     5701 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     5761 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5821 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5881 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5941 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6001 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6061 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6121 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6181 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6241 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6301 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6361 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6421 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6481 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6541 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gcattctagg
     6601 tactaaaaca attcatccag taaaatataa tattttattt tctcccaatc aggcttgatc
     6661 cccagtaagt caaaaaatag ctcgacatac tgttcttccc cgatatcctc cctgatcgac
     6721 cggacgcaga aggcaatgtc ataccacttg tccgccctgc cgcttctccc aagatcaata
     6781 aagccactta ctttgccatc tttcacaaag atgttgctgt ctcccaggtc gccgtgggaa
     6841 aagacaagtt cctcttcggg cttttccgtc tttaaaaaat catacagctc gcgcggatct
     6901 ttaaatggag tgtcttcttc ccagttttcg caatccacat cggccagatc gttattcagt
     6961 aagtaatcca attcggctaa gcggctgtct aagctattcg tatagggaca atccgatatg
     7021 tcgatggagt gaaagagcct gatgcactcc gcatacagct cgataatctt ttcagggctt
     7081 tgttcatctt catactcttc cgagcaaagg acgccatcgg cctcactcat gagcagattg
     7141 ctccagccat catgccgttc aaagtgcagg acctttggaa caggcagctt tccttccagc
     7201 catagcatca tgtccttttc ccgttccaca tcataggtgg tccctttata ccggctgtcc
     7261 gtcattttta aatataggtt ttcattttct cccaccagct tatatacctt agcaggagac
     7321 attccttccg tatcttttac gcagcggtat ttttcgatca gttttttcaa ttccggtgat
     7381 attctcattt tagccattta ttatttcctt cctcttttct atagtattta aagatacccc
     7441 aagaagctaa ttataacaag acgaactcca attcactgtt ccttgcattc taaaacctta
     7501 aataccagaa aacagctttt tcaaagttgt tttcaaagtt ggcgtataac atagtatcga
     7561 cggagccgat tttgaaaccg cggtgatcac aggcagcaac gctctgtcat cgttacaatc
     7621 aacatgctac cctccgcgag atcatccgtg tttcaaaccc ggcagcttag ttgccgttct
     7681 tccgaatagc atcggtaaca tgagcaaagt ctgccgcctt acaacggctc tcccgctgac
     7741 gccgtcccgg actgatgggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc
     7801 ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat tgacgcttag
     7861 acaacttaat aacacattgc ggacgttttt aatgtactga attaacgccg aattaattcg
     7921 ggggatctgg attttagtac tggattttgg ttttaggaat tagaaatttt attgatagaa
     7981 gtattttaca aatacaaata catactaagg gtttcttata tgctcaacac atgagcgaaa
     8041 ccctatagga accctaattc ccttatctgg gaactactca cacattatta tggagaaa
//
