LOCUS       40924_27054        6065 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Fungal transformation vector pKT300, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6065)
  AUTHORS   Twaruschek K, Spoerhase P, Michlmayr H, Wiesenberger G, Adam G.
  TITLE     New plasmids allowing positive-negative selection for fungal 
            transformation and efficient Cre-loxP mediated marker recycling
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6065)
  AUTHORS   Twaruschek K, Spoerhase P, Michlmayr H, Wiesenberger G, Adam G.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAY-2018) Department of Applied Genetics and Cell 
            Biology, University of Natural Resources and Life Sciences Vienna 
            (BOKU), Konrad Lorenz Strasse 24, Tulln 3430, Austria
REFERENCE   3  (bases 1 to 6065)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6065)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-MAY-2018) Department of Applied Genetics and Cell Biology, 
            University of Natural Resources and Life Sciences Vienna (BOKU), 
            Konrad Lorenz Strasse 24, Tulln 3430, Austria"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        363..467
                     /label=AmpR promoter
     CDS             468..1325
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1499..2087
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2345..2363
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2434..2467
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3233..4360
                     /codon_start=1
                     /label=HSV TK
                     /note="herpes simplex virus thymidine kinase"
                     /translation="MASYPCHQHASAFDQAARSRGHNNRRTALRPRRQQKATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGAAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     CDS             4400..5191
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(join(6064..6065,1..17))
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa ttgtagccgc
       61 gttctaacga caatatgtcc atatggtgca ctctcagtac aatctgctct gatgccgcat
      121 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
      181 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
      241 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
      301 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
      361 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      421 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      481 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      541 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
      601 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
      661 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
      721 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
      781 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
      841 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
      901 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
      961 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1021 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1081 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1141 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1201 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1261 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1321 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1381 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1441 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1501 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1561 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1621 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     1681 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1741 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1801 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1861 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1921 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1981 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2041 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2101 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2161 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2221 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     2281 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg caggttaacc tggcttatcg
     2341 aaattaatac gactcactat agggagaccg gcagatccgc ggccgcgttt aaactttaag
     2401 cttcatgatt taaatcagtc gacaaccctt aatataactt cgtataatgt atgctatacg
     2461 aagttattag gtctagatct cgagataacg gtgagactag cggccggtcc ccttatccca
     2521 gctgttccac gttggcctgc ccctcagtta gcgctcaact caatgcccct cactggcgag
     2581 gcgagggcaa ggatggaggg gcagcatcgc ctgagttgga gcaaagcggc cgggccgcca
     2641 tgggagcagc gaaccaacgg agggatgccg tgctttgtcg tggctgctgt ggccaatccg
     2701 ggcccttggt tggctcacag agcgttgctg tgagaccatg agctattatt gctaggtaca
     2761 gtatagagag aggagagaga gagagagaga gagagagggg aaaaaaggtg aggttgaagt
     2821 gagaaaaaaa aaaaaaaaaa aaaatccaac cactgacggc tgccggctct gccacccccc
     2881 tccctccacc ccagaccacc tgcacactca gcgcgcagca tcacctaatc ttggctcgcc
     2941 ttcccgcagc tcaggttgtt ttttttttct ctctccctcg tcgaagccgc ccttgttccc
     3001 ttatttattt ccctctccat ccttgtctgc ctttggtcca tctgcccctt tgtctgcatc
     3061 tcttttgcac gcatcgcctt atcgtcgtct cttttttcac tcacgggagc ttgacgaaga
     3121 cctgactcgt gagcctcacc tgctgatttc tctccccccc tcccgaccgg cttgactttt
     3181 gtttctcctc cagtacctta tcgcgaagcc ggaagaacct cttaacctct agatggcttc
     3241 gtacccctgc catcaacacg cgtctgcgtt cgaccaggct gcgcgttctc gcggccataa
     3301 caaccgacgt acggcgttgc gccctcgccg gcaacaaaaa gccacggaag tccgcctgga
     3361 gcagaaaatg cccacgctac tgcgggttta tatagacggt ccccacggga tggggaaaac
     3421 caccaccacg caactgctgg tggccctggg ttcgcgcgac gatatcgtct acgtacccga
     3481 gccgatgact tactggcggg tgttgggggc ttccgagaca atcgcgaaca tctacaccac
     3541 acaacaccgc ctcgaccagg gtgagatatc ggccggggac gcggcggtgg taatgacaag
     3601 cgcccagata acaatgggca tgccttatgc cgtgaccgac gccgttctgg ctcctcatat
     3661 cgggggggag gctgggagct cacatgcccc gcccccggcc ctcaccctca tcttcgaccg
     3721 ccatcccatc gccgccctcc tgtgctaccc ggccgcgcga taccttatgg gcagcatgac
     3781 cccccaggcc gtgctggcgt tcgtggccct catcccgccg accttgcccg gcacaaacat
     3841 cgtgttgggg gcccttccgg aggacagaca catcgaccgc ctggccaaac gccagcgccc
     3901 cggcgagcgg cttgacctgg ctatgctggc cgcgattcgc cgcgtttatg ggctgcttgc
     3961 caatacggtg cggtatctgc agggcggcgg gtcgtggcgg gaggattggg gacagctttc
     4021 gggggcggcc gtgccgcccc agggtgccga gccccagagc aacgcgggcc cacgacccca
     4081 tatcggggac acgttattta ccctgtttcg ggcccccgag ttgctggccc ccaacggcga
     4141 cctgtataac gtgtttgcct gggctttgga cgtcttggcc aaacgcctcc gtcccatgca
     4201 cgtctttatc ctggattacg accaatcgcc cgccggctgc cgggacgccc tgctgcaact
     4261 tacctccggg atggtccaga cccacgtcac caccccaggc tccataccga cgatctgcga
     4321 cctggcgcgc acgtttgccc gggagatggg ggaggctaac gaatgcagtg gaggaggttc
     4381 tggaaggagg atatgcataa tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4441 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4501 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4561 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4621 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4681 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4741 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4801 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4861 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4921 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4981 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5041 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5101 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5161 cgccttctat cgccttcttg acgagttctt ctgaggtacc gagctcgaat ttggctttcg
     5221 tgaccgggct tcaaacaatg atgtgcgatg gtgtggttcc cggttggcgg agtctttgtc
     5281 tactttggtt gtctgtcgca ggtcggtaga ccgcaaatga gcaactgatg gattgttgcc
     5341 agcgatacta taattcacat ggatggtctt tgtcgatcag tagctagtga gagagagaga
     5401 acatctatcc acaatgtcga gtgtctatta gacatactcc gagaataaag tcaactgtgt
     5461 ctgtgatcta aagatcgatt cggcagtcga gtagcgtata acaactccga gtaccagcga
     5521 aagcacgtcg tgacaggagc agggctttgc caactgcgca accttgcttg aatgaggata
     5581 cacggggtgc aacatggctg tactgatcca tcgcaaccaa aatttctgtt tatagatcaa
     5641 gctggtagat tccaattact ccacctcttg cgcttctcca tgacatgtaa gtgcacgtgg
     5701 aaaccatacc caaattgcct acagctgcgg agcatgagcc tatggcgatc agtctggtca
     5761 tgttaaccag cctgtgctct gacgttaatg cagaatagaa agccgcggtt gcaatgcaaa
     5821 tgatgatgcc tttgcagaaa tggcttgctc gctgactgat accagtaaca actttgcttg
     5881 gccgtctagc gctgttgatt gtattcatca caacctcgtc tccctccttt gggttgagct
     5941 ctcgagaacc cttaatataa cttcgtataa tgtatgctat acgaagttat taggtgatat
     6001 cagatccact agtatggcct atgcggccta gaattcgatt aattaaggat cctgcggccg
     6061 cgttc
//
