LOCUS       sequence-199530-        8494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for Agrobacterium tumefaciens-mediated transformation
            of Fusarium oxysporum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8494)
  AUTHORS   Mullins ED, Chen X, Romaine P, Raina R, Geiser DM, Kang S
  TITLE     Agrobacterium-Mediated Transformation of Fusarium oxysporum: An 
            Efficient Tool for Insertional Mutagenesis and Gene Transfer.
  JOURNAL   Phytopathology. 2001 Feb;91(2):173-80. doi: 
            10.1094/PHYTO.2001.91.2.173.
  PUBMED    18944391
REFERENCE   2  (bases 1 to 8494)
  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"; doi: 
            "10.1094/PHYTO.2001.91.2"; journalName: "Phytopathology"; date: 
            "2001-02"; volume: "91"; issue: "2"; pages: "173-80"
COMMENT     Sequence Label: pBHt2
FEATURES             Location/Qualifiers
     source          1..8494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    103..277
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(308..1333)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     protein_bind    1813..1834
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1849..1854,1855..1872,1873..1879)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1887..1903
                     /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     1892..1914
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1911..1927
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1911..1927
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1928..2155
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEC"
     misc_feature    1936..1992
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1996..2013)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1996..2012)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2005..2027)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    2215..2239
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3540..4169
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4598..5671
                     /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      5737..5931
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6167..6189)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6275..6415
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6330..6349
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6430..6447)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6601..7189)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6681..6700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7276..8070)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
       61 gacgttttta atgtactgaa ttaacgccga attaattcgg gggatctgga ttttagtact
      121 ggattttggt tttaggaatt agaaatttta ttgatagaag tattttacaa atacaaatac
      181 atactaaggg tttcttatat gctcaacaca tgagcgaaac cctataggaa ccctaattcc
      241 cttatctggg aactactcac acattattat ggagaaactc gaaactggtt cccggtcggc
      301 atctactcta ttcctttgcc ctcggacgag tgctggggcg tcggtttcca ctatcggcga
      361 gtacttctac acagccatcg gtccagacgg ccgcgcttct gcgggcgatt tgtgtacgcc
      421 cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc caagctgcat
      481 catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg cggagcatat
      541 acgcccggag gcgcggcgat cctgcaagct ccggatgcct ccgctcgaag tagcgcgtct
      601 gctgctccat acaagccaac cacggcctcc agaagaggat gttggcgacc tcgtattggg
      661 aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca ttgtccgtca
      721 ggacattgtt ggagccgaaa tccgcatgca cgaggtgccg gacttcgggg cagtcctcgg
      781 cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg tcgtccatca
      841 cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca cgccatgtag
      901 tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg ctaagatcgg
      961 ccgcagcgat cgcatccatg gcctccgcga ccggctggag aacagcgggc agttcggttt
     1021 caggcaggtc ttgcaacgtg acaccctgtg cacggcggga gatgcaatag gtcaggctct
     1081 cgctgaactc cccaatgtca agcacttccg gaatcgggag cgcggccgat gcaaagtgcc
     1141 gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc aggacatatc
     1201 cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag agctgcatca
     1261 ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc gcggtgagtt
     1321 caggcttttt catttggatg cttgggtaga ataggtaagt cagattgaat ctgaaataaa
     1381 gggaggaagg gcgaacttaa gaaggtatga ccgggtcgtc cacttacctt gcttgacaaa
     1441 cgcaccaagt tatcgtgcac caagcagcag atgataataa tgtcctcgtt cctgtctgct
     1501 aataagagtc acacttcgag cgccgccgct actgctacaa gtggggctga tctgaccagt
     1561 tgcctaaatg aaccatcttg tcaaacgaca caaattttgt gctcaccgcc tggacgacta
     1621 aaccaaaata ggcattcatt gttgacctcc actagctcca gccaagccca aaaaatgctc
     1681 cttcaatatc agttttggca agctgctcta gccaatacgc aaaccgcctc tccccgcgcg
     1741 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     1801 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     1861 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     1921 ctatgacatg attacgaatt cgagctcggt acccggggat cctctagagt cgacctgcag
     1981 gcatgcaagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2041 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2101 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gctagagcag
     2161 cttgagcttg gatcagattg tcgtttcccg ccttcagttt aaactatcag tgtttgacag
     2221 gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc ggatatttaa
     2281 aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc
     2341 ccctcgggat caaagtactt tgatccaacc cctccgctgc tatagtgcag tcggcttctg
     2401 acgttcagtg cagccgtctt ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca
     2461 ggctgccgcc ctgccctttt cctggcgttt tcttgtcgcg tgttttagtc gcataaagta
     2521 gaatacttgc gactagaacc ggagacatta cgccatgaac aagagcgccg ccgctggcct
     2581 gctgggctat gcccgcgtca gcaccgacga ccaggacttg accaaccaac gggccgaact
     2641 gcacgcggcc ggctgcacca agctgttttc cgagaagatc accggcacca ggcgcgaccg
     2701 cccggagctg gccaggatgc ttgaccacct acgccctggc gacgttgtga cagtgaccag
     2761 gctagaccgc ctggcccgca gcacccgcga cctactggac attgccgagc gcatccagga
     2821 ggccggcgcg ggcctgcgta gcctggcaga gccgtgggcc gacaccacca cgccggccgg
     2881 ccgcatggtg ttgaccgtgt tcgccggcat tgccgagttc gagcgttccc taatcatcga
     2941 ccgcacccgg agcgggcgcg aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc
     3001 taccctcacc ccggcacaga tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac
     3061 cgtgaaagag gcggctgcac tgcttggcgt gcatcgctcg accctgtacc gcgcacttga
     3121 gcgcagcgag gaagtgacgc ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc
     3181 attgaccgag gccgacgccc tggcggccgc cgagaatgaa cgccaagagg aacaagcatg
     3241 aaaccgcacc aggacggcca ggacgaaccg tttttcatta ccgaagagat cgaggcggag
     3301 atgatcgcgg ccgggtacgt gttcgagccg cccgcgcacg tctcaaccgt gcggctgcat
     3361 gaaatcctgg ccggtttgtc tgatgccaag ctggcggcct ggccggccag cttggccgct
     3421 gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt
     3481 catgcggtcg ctgcgtatat gatgcgatga gtaaataaac aaatacgcaa ggggaacgca
     3541 tgaaggttat cgctgtactt aaccagaaag gcgggtcagg caagacgacc atcgcaaccc
     3601 atctagcccg cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat tccgatcccc
     3661 agggcagtgc ccgcgattgg gcggccgtgc gggaagatca accgctaacc gttgtcggca
     3721 tcgaccgccc gacgattgac cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga
     3781 tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg
     3841 tgctgattcc ggtgcagcca agcccttacg acatatgggc caccgccgac ctggtggagc
     3901 tggttaagca gcgcattgag gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc
     3961 gggcgatcaa aggcacgcgc atcggcggtg aggttgccga ggcgctggcc gggtacgagc
     4021 tgcccattct tgagtcccgt atcacgcagc gcgtgagcta cccaggcact gccgccgccg
     4081 gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg
     4141 ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag
     4201 cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg caacgttggc
     4261 cagcctggca gacacgccag ccatgaagcg ggtcaacttt cagttgccgg cggaggatca
     4321 caccaagctg aagatgtacg cggtacgcca aggcaagacc attaccgagc tgctatctga
     4381 atacatcgcg cagctaccag agtaaatgag caaatgaata aatgagtaga tgaattttag
     4441 cggctaaagg aggcggcatg gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc
     4501 ccatgtgtgg aggaacgggc ggttggccag gcgtaagcgg ctgggttgcc tgccggccct
     4561 gcaatggcac tggaaccccc aagcccgagg aatcggcgtg agcggtcgca aaccatccgg
     4621 cccggtacaa atcggcgcgg cgctgggtga tgacctggtg gagaagttga aggccgcgca
     4681 ggccgcccag cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc
     4741 cgctgatcga atccgcaaag aatcccggca accgccggca gccggtgcgc cgtcgattag
     4801 gaagccgccc aagggcgacg agcaaccaga ttttttcgtt ccgatgctct atgacgtggg
     4861 cacccgcgat agtcgcagca tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg
     4921 acgagctggc gaggtgatcc gctacgagct tccagacggg cacgtagagg tttccgcagg
     4981 gccggccggc atggccagtg tgtgggatta cgacctggta ctgatggcgg tttcccatct
     5041 aaccgaatcc atgaaccgat accgggaagg gaagggagac aagcccggcc gcgtgttccg
     5101 tccacacgtt gcggacgtac tcaagttctg ccggcgagcc gatggcggaa agcagaaaga
     5161 cgacctggta gaaacctgca ttcggttaaa caccacgcac gttgccatgc agcgtacgaa
     5221 gaaggccaag aacggccgcc tggtgacggt atccgagggt gaagccttga ttagccgcta
     5281 caagatcgta aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc tagctgattg
     5341 gatgtaccgc gagatcacag aaggcaagaa cccggacgtg ctgacggttc accccgatta
     5401 ctttttgatc gatcccggca tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg
     5461 caaggcagaa gccagatggt tgttcaagac gatctacgaa cgcagtggca gcgccggaga
     5521 gttcaagaag ttctgtttca ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta
     5581 cgatttgaag gaggaggcgg ggcaggctgg cccgatccta gtcatgcgct accgcaacct
     5641 gatcgagggc gaagcatccg ccggttccta atgtacggag cagatgctag ggcaaattgc
     5701 cctagcaggg gaaaaaggtc gaaaaggtct ctttcctgtg gatagcacgt acattgggaa
     5761 cccaaagccg tacattggga accggaaccc gtacattggg aacccaaagc cgtacattgg
     5821 gaaccggtca cacatgtaag tgactgatat aaaagagaaa aaaggcgatt tttccgccta
     5881 aaactcttta aaacttatta aaactcttaa aacccgcctg gcctgtgcat aactgtctgg
     5941 ccagcgcaca gccgaagagc tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg
     6001 ccccgccgct tcgcgtcggc ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg
     6061 gccaggcaat ctaccagggc gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc
     6121 acatcaaggc accctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     6181 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     6241 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     6301 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6361 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     6421 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6481 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6541 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6601 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6661 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6721 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6781 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6841 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6901 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6961 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7021 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7081 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7141 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7201 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7261 catgcattct aggtactaaa acaattcatc cagtaaaata taatatttta ttttctccca
     7321 atcaggcttg atccccagta agtcaaaaaa tagctcgaca tactgttctt ccccgatatc
     7381 ctccctgatc gaccggacgc agaaggcaat gtcataccac ttgtccgccc tgccgcttct
     7441 cccaagatca ataaagccac ttactttgcc atctttcaca aagatgttgc tgtctcccag
     7501 gtcgccgtgg gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa aatcatacag
     7561 ctcgcgcgga tctttaaatg gagtgtcttc ttcccagttt tcgcaatcca catcggccag
     7621 atcgttattc agtaagtaat ccaattcggc taagcggctg tctaagctat tcgtataggg
     7681 acaatccgat atgtcgatgg agtgaaagag cctgatgcac tccgcataca gctcgataat
     7741 cttttcaggg ctttgttcat cttcatactc ttccgagcaa aggacgccat cggcctcact
     7801 catgagcaga ttgctccagc catcatgccg ttcaaagtgc aggacctttg gaacaggcag
     7861 ctttccttcc agccatagca tcatgtcctt ttcccgttcc acatcatagg tggtcccttt
     7921 ataccggctg tccgtcattt ttaaatatag gttttcattt tctcccacca gcttatatac
     7981 cttagcagga gacattcctt ccgtatcttt tacgcagcgg tatttttcga tcagtttttt
     8041 caattccggt gatattctca ttttagccat ttattatttc cttcctcttt tctacagtat
     8101 ttaaagatac cccaagaagc taattataac aagacgaact ccaattcact gttccttgca
     8161 ttctaaaacc ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat
     8221 aacatagtat cgacggagcc gattttgaaa ccgcggtgat cacaggcagc aacgctctgt
     8281 catcgttaca atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct
     8341 tagttgccgt tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg
     8401 ctctcccgct gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc
     8461 cgagctgccg gtcggggagc tgttggctgg ctgg
//
