LOCUS       sequence-337339-        8089 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A. tumefaciens TI plasmid encoding NATca in T-DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8089)
  AUTHORS   Santana DJ, O'Meara TR
  TITLE     Forward and reverse genetic dissection of morphogenesis identifies 
            filament-competent Candida auris strains.
  JOURNAL   Nat Commun. 2021 Dec 10;12(1):7197. doi: 10.1038/s41467-021-27545-5.
  PUBMED    34893621
REFERENCE   2  (bases 1 to 8089)
  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: "Nat 
            Commun."; date: "2021-12-10"; volume: "12(1):7197. doi"; pages: " 
            10.1038/s41467-021-27545-5"
COMMENT     Sequence Label: pTO128_pPZP-NATca
FEATURES             Location/Qualifiers
     source          1..8089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1333..1962
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2391..3464
                     /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      3530..3724
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(3960..3982)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4068..4208
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4123..4142
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4223..4240)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4394..4982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4474..4493)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5077..5892)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5077..5892)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5800..5819
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5884..5903
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     misc_feature    6185..6209
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    6386..6407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6422..6427,6428..6445,6446..6452)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6460..6476
                     /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     6465..6487
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6484..6500
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6484..6500
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6553..6569
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6553..6569
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    6571..6604
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        6645..6988
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      7589..7786
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(7855..7872)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7855..7871)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7864..7886)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    join(8067..8089,1..2)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
       61 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta
      121 ctttaaagta ctttaaagta ctttaaagta ctttgatcca acccctccgc tgctatagtg
      181 cagtcggctt ctgacgttca gtgcagccgt cttctgaaaa cgacatgtcg cacaagtcct
      241 aagttacgcg acaggctgcc gccctgccct tttcctggcg ttttcttgtc gcgtgtttta
      301 gtcgcataaa gtagaatact tgcgactaga accggagaca ttacgccatg aacaagagcg
      361 ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga cgaccaggac ttgaccaacc
      421 aacgggccga actgcacgcg gccggctgca ccaagctgtt ttccgagaag atcaccggca
      481 ccaggcgcga ccgcccggag ctggccagga tgcttgacca cctacgccct ggcgacgttg
      541 tgacagtgac caggctagac cgcctggccc gcagcacccg cgacctactg gacattgccg
      601 agcgcatcca ggaggccggc gcgggcctgc gtagcctggc agagccgtgg gccgacacca
      661 ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg cattgccgag ttcgagcgtt
      721 ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc caaggcccga ggcgtgaagt
      781 ttggcccccg ccctaccctc accccggcac agatcgcgca cgcccgcgag ctgatcgacc
      841 aggaaggccg caccgtgaaa gaggcggctg cactgcttgg cgtgcatcgc tcgaccctgt
      901 accgcgcact tgagcgcagc gaggaagtga cgcccaccga ggccaggcgg cgcggtgcct
      961 tccgtgagga cgcattgacc gaggccgacg ccctggcggc cgccgagaat gaacgccaag
     1021 aggaacaagc atgaaaccgc accaggacgg ccaggacgaa ccgtttttca ttaccgaaga
     1081 gatcgaggcg gagatgatcg cggccgggta cgtgttcgag ccgcccgcgc acgtctcaac
     1141 cgtgcggctg catgaaatcc tggccggttt gtctgatgcc aagctggcgg cctggccggc
     1201 cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa aggtgatgtg tatttgagta
     1261 aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga tgagtaaata aacaaatacg
     1321 caaggggaac gcatgaaggt tatcgctgta cttaaccaga aaggcgggtc aggcaagacg
     1381 accatcgcaa cccatctagc ccgcgccctg caactcgccg gggccgatgt tctgttagtc
     1441 gattccgatc cccagggcag tgcccgcgat tgggcggccg tgcgggaaga tcaaccgcta
     1501 accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg tgaaggccat cggccggcgc
     1561 gacttcgtag tgatcgacgg agcgccccag gcggcggact tggctgtgtc cgcgatcaag
     1621 gcagccgact tcgtgctgat tccggtgcag ccaagccctt acgacatatg ggccaccgcc
     1681 gacctggtgg agctggttaa gcagcgcatt gaggtcacgg atggaaggct acaagcggcc
     1741 tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg gtgaggttgc cgaggcgctg
     1801 gccgggtacg agctgcccat tcttgagtcc cgtatcacgc agcgcgtgag ctacccaggc
     1861 actgccgccg ccggcacaac cgttcttgaa tcagaacccg agggcgacgc tgcccgcgag
     1921 gtccaggcgc tggccgctga aattaaatca aaactcattt gagttaatga ggtaaagaga
     1981 aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc gagcgcacgc agcagcaagg
     2041 ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa gcgggtcaac tttcagttgc
     2101 cggcggagga tcacaccaag ctgaagatgt acgcggtacg ccaaggcaag accattaccg
     2161 agctgctatc tgaatacatc gcgcagctac cagagtaaat gagcaaatga ataaatgagt
     2221 agatgaattt tagcggctaa aggaggcggc atggaaaatc aagaacaacc aggcaccgac
     2281 gccgtggaat gccccatgtg tggaggaacg ggcggttggc caggcgtaag cggctgggtt
     2341 gtctgccggc cctgcaatgg cactggaacc cccaagcccg aggaatcggc gtgagcggtc
     2401 gcaaaccatc cggcccggta caaatcggcg cggcgctggg tgatgacctg gtggagaagt
     2461 tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc cccggtgaat
     2521 cgtggcaagc ggccgctgat cgaatccgca aagaatcccg gcaaccgccg gcagccggtg
     2581 cgccgtcgat taggaagccg cccaagggcg acgagcaacc agattttttc gttccgatgc
     2641 tctatgacgt gggcacccgc gatagtcgca gcatcatgga cgtggccgtt ttccgtctgt
     2701 cgaagcgtga ccgacgagct ggcgaggtga tccgctacga gcttccagac gggcacgtag
     2761 aggtttccgc agggccggcc ggcatggcca gtgtgtggga ttacgacctg gtactgatgg
     2821 cggtttccca tctaaccgaa tccatgaacc gataccggga agggaaggga gacaagcccg
     2881 gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt ctgccggcga gccgatggcg
     2941 gaaagcagaa agacgacctg gtagaaacct gcattcggtt aaacaccacg cacgttgcca
     3001 tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac ggtatccgag ggtgaagcct
     3061 tgattagccg ctacaagatc gtaaagagcg aaaccgggcg gccggagtac atcgagatcg
     3121 agctagctga ttggatgtac cgcgagatca cagaaggcaa gaacccggac gtgctgacgg
     3181 ttcaccccga ttactttttg atcgatcccg gcatcggccg ttttctctac cgcctggcac
     3241 gccgcgccgc aggcaaggca gaagccagat ggttgttcaa gacgatctac gaacgcagtg
     3301 gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg caagctgatc gggtcaaatg
     3361 acctgccgga gtacgatttg aaggaggagg cggggcaggc tggcccgatc ctagtcatgc
     3421 gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc ctaatgtacg gagcagatgc
     3481 tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg tctctttcct gtggatagca
     3541 cgtacattgg gaacccaaag ccgtacattg ggaaccggaa cccgtacatt gggaacccaa
     3601 agccgtacat tgggaaccgg tcacacatgt aagtgactga tataaaagag aaaaaaggcg
     3661 atttttccgc ctaaaactct ttaaaactta ttaaaactct taaaacccgc ctggcctgtg
     3721 cataactgtc tggccagcgc acagccgaag agctgcaaaa agcgcctacc cttcggtcgc
     3781 tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc ggccgctggc cgctcaaaaa
     3841 tggctggcct acggccaggc aatctaccag ggcgcggaca agccgcgccg tcgccactcg
     3901 accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     3961 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4021 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     4081 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     4141 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     4201 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4261 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4321 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4381 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4441 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4501 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4561 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4621 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4681 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4741 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4801 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4861 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4921 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4981 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5041 aagggatttt ggtcatgcag gatcatgaat taattcttag aaaaactcat cgagcatcaa
     5101 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     5161 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     5221 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     5281 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     5341 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     5401 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     5461 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     5521 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     5581 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     5641 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     5701 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     5761 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     5821 atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg
     5881 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     5941 tgatctggat cacaggcagc aacgctctgt catcgttaca atcaacatgc taccctccgc
     6001 gagatcatcc gtgtttcaaa cccggcagct tagttgccgt tcttccgaat agcatcggta
     6061 acatgagcaa agtctgccgc cttacaacgg ctctcccgct gacgccgtcc cggactgatg
     6121 ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc tgttggctgg
     6181 ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt aataacacat
     6241 tgcggacgtt tttaatgtac tgaattaacg ccgaattgct ctagccaata cgcaaaccgc
     6301 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     6361 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     6421 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     6481 acacaggaaa cagctatgac atgattacga attcttaatt aagatatcga gctccaccgc
     6541 ggtggcggcc gctctagaac tagtggatct gaagttccta ttctctagaa agtataggaa
     6601 cttccatcaa gcttgcctcg tccccgccgg gtcacccggc cagcgacatg gaggcccaga
     6661 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
     6721 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
     6781 cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga aacgctcccc
     6841 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
     6901 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
     6961 ctcacatcac atccgaacat aaacaaccgg atcctacaat tatgtctact actttggatg
     7021 atactgctta tagatacaga acttctgttc caggtgatgc tgaagctatt gaagctttgg
     7081 atggttcttt cactaccgat actgttttta gagttactgc tactggtgat ggtttcactt
     7141 tgagagaagt tccagttgat ccaccattga ctaaggtttt cccagatgat gaatccgatg
     7201 atgaatccga tgctggtgaa gatggtgatc cagattctag aactttcgtt gcttatggtg
     7261 atgatggtga tttggctggt ttcgttgttg tttcttattc tggttggaac agaagattga
     7321 ctgttgaaga tattgaagtt gctccagaac atagaggtca tggtgttggt agagctttga
     7381 tgggtttggc tactgaattc gccagagaaa gaggtgctgg tcatttgtgg ttggaagtta
     7441 ccaatgttaa tgctccagct attcatgctt atagaagaat gggtttcact ttgtgtggtt
     7501 tggatactgc tttatacgat ggtactgctt ccgatggtga acaagctttg tatatgtcta
     7561 tgccatgtcc ataataactc gagtcagtac tgacaataaa aagattcttg ttttcaagaa
     7621 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
     7681 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
     7741 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata
     7801 ctaacgccgc catccagtgt cgacctgcag aggcctaggc gcgccaagct tggcactggc
     7861 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     7921 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     7981 ccaacagttg cgcagcctga atggcgaatg agcttgagct tggatcagat tgtcgtttcc
     8041 cgccttcagt ttaaactatc agtgtttgac aggatatatt ggcgggtaa
//
