LOCUS       sequence-251697-        8576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing synthetic TetR-GFP protein in N. crassa (from the his-3 
            locus).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8576)
  AUTHORS   Nguyen TS, Gladyshev E
  TITLE     Developing a tetO/TetR system in Neurospora crassa.
  JOURNAL   Fungal Genet Biol. 2020 Mar;136:103316. doi: 
            10.1016/j.fgb.2019.103316. Epub 2019 Dec 9.
  PUBMED    31821884
REFERENCE   2  (bases 1 to 8576)
  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: "Fungal 
            Genet Biol."; date: "2020-03"; volume: "136:103316. doi"; pages: " 
            10.1016/j.fgb.2019.103316. Epub 2019 Dec 9"
COMMENT     Sequence Label: pTSN7A
FEATURES             Location/Qualifiers
     source          1..8576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(98..117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     217..239
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(277..295)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        363..467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(468..536,537..1328)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(686..705)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1499..2087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1988..2007
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2241..2258
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2345..2363
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2638..2649
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4008..4026
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(5110..5112,5113..5115,5116..5829)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5155..5176)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5416..5435)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5763..5784
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5850..5866)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5851..5867)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5896..5914)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(join(8575..8576,1..17))
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(join(8576,1..17))
                     /label=SP6
                     /note="SP6 promoter, forward primer"
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 attgagaaag 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 gcagatccaa tgcggatgga ttcgctcact
     2401 cggtgccgcg gcggtgcgaa gttcctgccg tggattgggg gattgcacag acggtggagt
     2461 gcttgctaca gctaggtacc cagttgcaca ccaaacggtg cccctgtgcc cctgcgtcta
     2521 ggtgtgtcct gtgagcaact gcgcccgcgc gcggcaccgg gaagctgcgg gaacctcaat
     2581 ggatgattgg cgccagatcc ccggcatcac ctcagcacat gggcagcccg agaaggaatc
     2641 gagggtagaa gatattccag ggtttccgct agcacaactt ccaagcgcct ctaccgctac
     2701 tccacacacc cacgctgaat ggcacaacca ctgtaggcct agcggtccca gctccccgta
     2761 accacgcagc cccagcgacc gaggtcatca tgacactatg acgacaatct ggcgggagac
     2821 ttgggaaaat cgatgacggg cacctacctt gggaacaata gagcaagtcc agtcaacgcc
     2881 ctagggaagg gggaaaaaaa aaaagagccc attctcacaa ggagcaacac tgtgctgcta
     2941 tcatagtaca ctacaagcaa agtcgggatg ttgctagaat ccgtcggcta cgtcggtaga
     3001 tacctcccat aggcagtaag tctaggtatc caatttatag ttccgcgtat ttcttcgtcg
     3061 aacggggcac agcaaagcct aacagccgca gtgcctcgca accaacacca agcagtaatc
     3121 tcatccatcc atacagatcc tcccacaata tctttacatg cgcacttgcg ctgtgttagc
     3181 agtttaggtt ccactaccaa ggctgacacg agcagcactg acagcaaact gctgcttggg
     3241 gccaagaaat tggcagtttg ctgagcacct acatatcagt acacatcaac cctggtttaa
     3301 ttgttagcct ggtactgtta accgtcatga acgaggacgt ggaggaagaa ccacgctaac
     3361 aattgaacca gggttgatgt atacctctct taccatattt gcttgcttcc acgtccctat
     3421 cctcaagcga ttttgttctt gactttttgt atttcaacgc aaaggccatt caggtttctc
     3481 atgttcaacc ctttggatgg cgatgccgtt cgtcccagga cctcgtgacc tttgaaagac
     3541 gagaaatata tcccaaagca acttctgctt atggacaagc aactgcatgg cggcccacaa
     3601 ctcatatctc gcgtaatcat ccaaacgcta ggcccgagta ctctactgtt ctcgagacac
     3661 ctaccataat atgtacccaa tcagacatga caacagtcaa ccaaacatac cctgatccgg
     3721 ccaaaccctg acatccctag atgatggtta ggcacacaaa tggaaaacgg gactcgttgg
     3781 ttaggtggga acgcttgtat tgcagccagc caaccgactc tcctcgtccc tctcgagtcc
     3841 cgttattgcc gtttgaccca gccgcccagc cgccaaacag ctgagggagc caatggtcta
     3901 atacaccgtc caacggcccc agtgatgttt caaacttgat tgacagcgaa cgaaacccct
     3961 gaaacccgtt atagatatgt atagatttga cggggtagct tggcccttaa tacgactcac
     4021 tatagggcga attggagctc caccgcggtg gcggccgcag aagatgatat tgaaggagca
     4081 ctttttgggc ttggctggag ctagtggagg tcaacaatga atgcctattt tggtttagtc
     4141 gtccaggcgg tgagcacaaa atttgtgtcg tttgacaaga tggttcattt aggcaactgg
     4201 tcagatcagc cccacttgta gcagtagcgg cggcgctcga agtgtgactc ttattagcag
     4261 acaggaacga ggacattatt atcatctgct gcttggtgca cgataacttg gtgcgtttgt
     4321 caagcaaggt aagtgaacga cccggtcata ccttcttaag ttcgcccttc ctccctttat
     4381 ttcagattca atctgactta cctattctac cgaagctcta gaatggccat gtcccgcctc
     4441 gacaagtcca aggtcatcaa ctccgctctc gagctcctca acgaggtcgg tatcgagggt
     4501 ctcaccaccc gcaagctcgc tcagaagctc ggtgtcgagc agcctaccct ctactggcac
     4561 gtcaagaaca agcgcgctct cctcgacgct ctcgccatcg agatgctcga ccgccaccac
     4621 acccacttca gccctctcga gggtgagtcc tggcaggact tcctccgcaa caacgccaag
     4681 tccttccgca acgctctgct ctcccaccgc gacggtgcta aggtccacct cggtacccgc
     4741 cctaccgaga agcagtacga gaccctcgag aaccagctcg ctttcctcac ccagcagggc
     4801 ttctccctcg agaacgccct ctacgctctc tccgctgtcg gtcacttcac cctcggctcc
     4861 gtcctcgagg accaggagca ccaggtcgcc aaggaggaac gcgagacccc taccaccgac
     4921 tccatgcctc ctctcctccg ccaggccatc gagctcttcg accaccaggg tgccgagcct
     4981 gccttcctcc acggcctcga gtccctcatc cgcggtttcg aggtccagct caccgctctc
     5041 ctccagatcg tctccggtag cggcttaatc agcgctccta ggcgcaagaa gcgcaagacc
     5101 ggtattaaca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     5161 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     5221 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     5281 tggcccaccc tcgtgaccac cttcacctac ggcgtgcagt gcttcagccg ctaccccgac
     5341 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     5401 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     5461 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     5521 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     5581 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     5641 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     5701 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     5761 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     5821 tacaagtaag aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt
     5881 acccagcttt tgttcccttt agtgagggtt aattagggcg tggacggcta atggggtctg
     5941 aatgctaaaa gacaccattt cccacactcc ctctgatttt ttttatggtt tttttttatc
     6001 ttcatttttc gccgtctttt attcggttgt ttgggaggtt cacatgcaac gtgcaagact
     6061 gcaagttttt tgagatgggc gcttcatgca atgcagcatc actgcaggga cattgtgaca
     6121 tccacgtttc gatcgtctgt gcttcccccg ttcatgtacg catgatctga aaaaaaaaaa
     6181 aaaaaaaaag gttgatgtaa taatgaaaat aaacacccag ggacgagcac ctcatctgct
     6241 ggaataggga gaaaaactca agggccgttt gggtctccac acttaccaac agattcatgt
     6301 actgatcttc agcctgtaca gaacagtaca atgtccaaaa ctcgttatca ttactctatg
     6361 tgcccaaaat gacccagata tacatcaacc cactacatcc ttttgttctt gttttttttt
     6421 tttttttttg ctacttcgaa aagaatttcc gtttagttgc tcttgctcat gtgctcaaga
     6481 cggatgctga ccgcccttct gtgagcctcg agctcctcaa ccttagccaa ctgcatgaca
     6541 gcctggccga cgtttttgag accctcggca gtcaagttgg agctggtaat gtgcttgacg
     6601 aacgaggcga aattgacgcc agagtactgc ttgccaaagc cataggtagc tatgtgtctt
     6661 gacgttagca gaagaatctg tgaagcgggg ctatttatga tatgtaactt acgcagcgag
     6721 tggttaacac cagcagagta atcgccaacg gactcaggag tccaggcgcc aatgaagaca
     6781 ctaccggcgt tcatgacaag atcgacagcc ttctcggcct ccttgatctg gaggatcaag
     6841 tgctcaggag cgtacttgtt gctgagctcc atggcctcct cgacggtctt gacctgcacg
     6901 gtgatcgagt gggcgatgga gccacggaca atctggacgc gaggaagctc cgtagcctga
     6961 cggtgaacct cgtcctcaat agcctgaaga tgctcctcgt cgaggtcaat agcgatcagg
     7021 atgacctgac tgtcaacgcc gtgctcagcc tgggacagga gatccgaggc aacgaacgcg
     7081 gggttggcgt ccttgtcagc gatgaccagc acctcggacg ggccagcggg catgtcaata
     7141 ccaacggcag cgttggtgtc gttgctgacg aacatcttgg cagcagtgac gaactggtta
     7201 ccggggccga gaatcttgtc gaccttggtg atgctctcgg tgccgtaggc catggcagct
     7261 acggcctggg caccgccggc aagcacgatg gactcggccc caaccttgtg agcgacgtgg
     7321 acaatctcgg gagtgatggt tccgtcggcg cggggaggag aggcgaacac aatcttgttg
     7381 cagccggcga ccatggcggg aacacccagc ataagggcag tgctggggag aacggcggta
     7441 ccgccgggga tgtagcagcc gacggcctcg atgggacgag agaaacggct gcagacaaca
     7501 ccgggcatgg tctcgacctg gaggggcttc tcctccttct gggcggcgtg gaacttgcgg
     7561 atgttctcga aggacacgtc gatggcagca atggtctcct cagggagctg cataagctcc
     7621 ttggggaagg gcgccttcag gacggggcta gtaagagagg tagccttctc gaacttgtga
     7681 gtgtacgaca gaacagcctt gtcgccgttc ttgcggacgt cctcgatgat ggggacaatg
     7741 atcttgtaga tggcatcgga cgacttttgc gcaggacgct tgagagcagc atcgagctcc
     7801 tcggtagaga ccttggaggc gtcgaaacgt ctcatggtga tcttctcagg ggtggtctcc
     7861 tgggcggcct ccttggtgac aggggccgaa gtggcagcga gagcggacgc cgcaggcttg
     7921 atgccctcct tctcagccca cttaccctta gcatctccag tcctgcgctt gaccttccag
     7981 ctcttggcgt caaggctcct ttcgatatcg gcaagagtaa cgccggcagc aacggccctg
     8041 gtaagagcaa agtagaagag atcggcagcc tcaaaggcga tttcctgggg ggtctgagcg
     8101 gtgcagagct cctcagcctc ctccatgatc ttggcccgga ctagcttctc atcggagaag
     8161 agacgggcag tgtaggagcc ctcgggggca gactgtttcc tcgaaatcaa agtctgctcg
     8221 agcttgggaa ggcctttgag ctgaccaaag cagccggact gatcgaggtg gcagaaacga
     8281 cccttctgct tcacgacaaa cttgagagca tcgttatcgc agtcaagcga gatgcggacg
     8341 agctcctgag tgtctccgga agtagcaccc ttgtaccaga gaccgcgctt ccggctctga
     8401 tagacaccag tctgtgtcct gagggcctcg ttcacactct cggcactgct gtagaccaga
     8461 cccagagcag tgtcgtgctc atcaacgaca acggtgggga gaagaccatc gggacggtca
     8521 gacttccaga cgctcgagag gatggtggag atggcaagct tcagctgctc gagttc
//
