LOCUS       sequence-225405-        4571 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  TetOFF controlled GFP-tagging vector with Hygromycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4571)
  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 4571)
  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: pFGL1252_TetGFP(Hyg)
FEATURES             Location/Qualifiers
     source          1..4571
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             628..648
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      888..1140
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(1790..2815)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     protein_bind    3480..3498
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    3686..3704
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             join(3849..3851,3852..3854,3855..4565)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3849..3851,3852..3854,3855..4565)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3894..3915)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4155..4174)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4502..4523
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gaattcatgg cccgcctgga taagtcaaag gtcatcaact ctgccttgga gctgctgaac
       61 gaggtcggca tcgagggtct cacaacgagg aagctggccc agaagctggg tgtcgagcag
      121 ccaaccctct actggcacgt caagaacaag cgcgccctcc tggacgccct cgccatcgag
      181 atgctggacc gccaccacac ccacttctgc cccctggagg gcgagagctg gcaggacttc
      241 ctgcgcaaca acgccaagag cttcaggtgc gccctcctga gccacaggga cggcgccaag
      301 gtccacctcg gcacccgccc caccgagaag cagtacgaga ccctggagaa ccagctcgcc
      361 ttcctgtgcc agcagggctt cagcctggag aacgccctct acgccctgag cgccgtcggt
      421 cacttcaccc tgggttgcgt cctggaggac caggagcacc aggtcgccaa ggaagagcgc
      481 gagaccccga ccaccgacag catgcccccg ctcctgcgcc aggccatcga gctgttcgac
      541 caccagggtg ccgagcccgc cttcctgttc ggccttgagc ttatcatctg cggcctggag
      601 aagcagctga agtgcgagag cggctcgccc aagaagaagc gcaaggtcga gggcagcgtc
      661 agcatgcagc caagcctcag gagcgagtac gagtacccgg tcttcagcca cgtccaggcc
      721 ggcatgttca gccccgagct gcgcaccttc accaagggtg acgccgagcg ctgggtcagc
      781 ggctcgccgg ccgacgccct ggatgatttc gacctggata tgctccctgc cgacgccctg
      841 gacgacttcg acctggatat gctgtgaggc gcggccggcc gcccggcgat cgttcaaaca
      901 tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat
      961 aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta
     1021 tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca
     1081 aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc
     1141 cgatgataag ctgtcaaaca tgagaattgc ggccgcgtcg acaagaagga ttacctctaa
     1201 acaagtgtac ctgtgcattc tgggtaaacg actcatagga gagttgtaaa aaagtttcgg
     1261 ccggcgtatt gggtgttacg gagcattcac taggcaacca tgcatggtta ctattgtata
     1321 cccatcttag taggaatgat tttcgaggtt tatacctacg atgaatgtgt gtcctgtagg
     1381 cttgagagtt caaggaagaa acagtgcaat tatctttgcg aacccagggg ctggtgacgg
     1441 aattttcata gtcaagctat cagagtaaag aagaggagca tgtcaaagta caattagaga
     1501 caaatatata gtcgcgtgga gccaagagcg gattcctcag tctcgtaggt ctcttgacga
     1561 ccgttgatct gcttgatctc gtctcccgaa aatgaaaata gactctgcta agctattctt
     1621 ctgcttcgcc ggagcctgaa gggcgtacta gggttgcgag gtccaatgca ttaatgcatt
     1681 gcagatgagc tgtatctgga agaggtaaac ccgaaacgca ttttattctt gttgacatgg
     1741 agctattaaa tcactagaat tactcgactg gttccggtcg gcatctactc tatttctttg
     1801 ccctcggacg agtgctgggg cgtcggtttc cactatcggc gagtacttct acacagccat
     1861 cggtccagac ggccgcgctt ctgcgggcga tttgtgtacg cccgacagtc ccggctccgg
     1921 atcggacgat tgcgtcgcat cgaccctgcg cccaagctgc atcatcgaaa ttgccgtcaa
     1981 ccaagctctg atagagttgg tcaagaccaa tgcggagcat atacgcccgg agtcgtggcg
     2041 atcctgcaag ctccggatgc ctccgctcga agtagcgcgt ctgctgctcc atacaagcca
     2101 accacggcct ccagaagaag atgttggcga cctcgtattg ggaatccccg aacatcgcct
     2161 cgctccagtc aatgaccgct gttatgcggc cattgtccgt caggacattg ttggagccga
     2221 aatccgcgtg cacgaggtgc cggacttcgg ggcagtcctc ggcccaaagc atcagctcat
     2281 cgagagcctg cgcgacggac gcactgacgg tgtcgtccat cacagtttgc cagtgataca
     2341 catggggatc agcaatcgcg catatgaaat cacgccatgt agtgtattga ccgattcctt
     2401 gcggtccgaa tgggccgaac ccgctcgtct ggctaagatc ggccgcagcg atcgcatcca
     2461 tagcctccgc gaccggttgt agaacagcgg gcagttcggt ttcaggcagg tcttgcaacg
     2521 tgacaccctg tgaacggcgg gagatgcaat aggtcaggct ctcgctaaac tccccaatgt
     2581 caagcacttc cggaatcggg agcgcggccg atgcaaagtg ccgataaaca taacgatctt
     2641 tgtagaaacc atcggcgcag ctatttaccc gcaggacata tccacgccct cctacatcga
     2701 agctgaaagc acgagattct tcgccctccg agagctgcat caggtcggag acgctgtcga
     2761 acttttcgat cagaaacttc tcgacagacg tcgcggtgag ttcaggcttt ttcatttgga
     2821 tgcttgggta gaataggtaa gtcagattga atctgaaata aagggaggaa gggcgaactt
     2881 aagaaggtat gaccgggtcg tccacttacc ttgcttgaca aacgcaccaa gttatcgtgc
     2941 accaagcagc agatgataat aatgtcctcg ttcctgtctg ctaataagag tcacacttcg
     3001 agcgccgccg ctactgctac aagtggggct gatctgacca gttgcctaaa tgaaccatct
     3061 tgtcaaacga cacaaatttt gtgctcaccg cctggacgac taaaccaaaa taggcattca
     3121 ttgttgacct ccactagctc cagccaagcc caaaaagtgc tccttcaata tcctcgactc
     3181 tagataaatc attccactca acattcaggc tcctctgcgc acgtaaagtg ccaaaggcaa
     3241 taccctgctc ggtggaatgc cgccgggctt gtcgatttta cgcacatagg cgcattcttg
     3301 acttgaagcg gaggagttct tcgttgcggg ttacagtgtt ttaataaaag aatggtcaaa
     3361 tcaaactgct agatatacct gtcagacact ctagttgttg acccctatac tcttaataca
     3421 tcagacagta catgcatgtt gcatgatgat gtcggtacaa ctcccactag acatatccta
     3481 ctctatcatt gatagagtga cccttccata gacatatcct actctatcat tgatagattg
     3541 acccttccac taggttcact tttctctatc attgataggg agtggtaaac aattgatgga
     3601 caaaatctat caatgataga gagtgcatgt tacctagtat tgttcttctc tatcattgat
     3661 agtgactttg cctaatcact tagcaactct atcattgata gagttctagt attagcaagg
     3721 cctttcctca gatcccgatc aatccgacag ccaaacctca tccactctca ctttcacact
     3781 ctaacttata cgatcacttc tcgcccgttc ttttgaacat caaatcaact accttacttc
     3841 tatcaattat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     3901 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     3961 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     4021 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     4081 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     4141 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     4201 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     4261 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     4321 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     4381 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     4441 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     4501 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     4561 acaagggtac c
//
