LOCUS       sequence-255822-        8569 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 8569)
  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 8569)
  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..8569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1058..1060,1061..1063,1064..1777)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1103..1124)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1364..1383)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1711..1732
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1798..1814)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1799..1815)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4517..4535)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4518..4535)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4616..4635)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4735..4757
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4795..4813)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4881..4985
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4986..5054,5055..5846)
                     /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(5204..5223)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6017..6605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6506..6525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6759..6776
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6863..6881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7156..7167
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        8525..8543
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggccgcagaa gatgatattg aaggagcact ttttgggctt ggctggagct agtggaggtc
       61 aacaatgaat gcctattttg gtttagtcgt ccaggcggtg agcacaaaat ttgtgtcgtt
      121 tgacaagatg gttcatttag gcaactggtc agatcagccc cacttgtagc agtagcggcg
      181 gcgctcgaag tgtgactctt attagcagac aggaacgagg acattattat catctgctgc
      241 ttggtgcacg ataacttggt gcgtttgtca agcaaggtaa gtgaacgacc cggtcatacc
      301 ttcttaagtt cgcccttcct ccctttattt cagattcaat ctgacttacc tattctaccg
      361 aagctctaga atggccatgt cccgcctcga caagtccaag gtcatcaact ccgctctcga
      421 gctcctcaac gaggtcggta tcgagggtct caccacccgc aagctcgctc agaagctcgg
      481 tgtcgagcag cctaccctct actggcacgt caagaacaag cgcgctctcc tcgacgctct
      541 cgccatcgag atgctcgacc gccaccacac ccacttcagc cctctcgagg gtgagtcctg
      601 gcaggacttc ctccgcaaca acgccaagtc cttccgcaac gctctgctct cccaccgcga
      661 cggtgctaag gtccacctcg gtacccgccc taccgagaag cagtacgaga ccctcgagaa
      721 ccagctcgct ttcctcaccc agcagggctt ctccctcgag aacgccctct acgctctctc
      781 cgctgtcggt cacttcaccc tcggctccgt cctcgaggac caggagcacc aggtcgccaa
      841 ggaggaacgc gagaccccta ccaccgactc catgcctcct ctcctccgcc aggccatcga
      901 gctcttcgac caccagggtg ccgagcctgc cttcctccac ggcctcgagt ccctcatccg
      961 cggtttcgag gtccagctca ccgctctcct ccagatcgtc tccggtagcg gcttaatcag
     1021 cgctcctagg cgcaagaagc gcaagaccgg tattaacatg gtgagcaagg gcgaggagct
     1081 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1141 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1201 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcacctacgg
     1261 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1321 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1381 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1441 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1501 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1561 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1621 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     1681 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1741 cgggatcact cacggcatgg acgagctgta caagtaagaa ttcgatatca agcttatcga
     1801 taccgtcgac ctcgaggggg ggcccggtac cagcttttgt tccctttagt gagggttaat
     1861 agggcgtgca cggctatggg gtctgaatgc taaaagacac catttcccac actccctctg
     1921 atttttttta tggttttttt ttatcttcat ttttcgccgt cttttattcg gttgtttggg
     1981 aggttcacat gcaacgtgca agactgcaag tatattgaga tgggcgcttc atgcaatgca
     2041 gcatcactgc agcgacattg tgacatccac gtttcgatcg tctgtgcttc cccgttcatg
     2101 tacgcatgat ctgaaaaaaa aaaaaaaaaa aaaaaagttg atgtaataat gaaaataaac
     2161 acccagggac gagcacccct tctggaatag ggagaaaaac taaagggacg tttgggtctc
     2221 tacacttacc aacagattca tgtactgatc ttcagcctgt acagaacagt acaatgtcca
     2281 aaactcgtta tcattactct atgtgcccaa aatgacccag atatacatca acccactaca
     2341 tccttttgtt cttgtttttt tttttttttt ttgctacttc gaaaagaatt tccgtttagt
     2401 tgctcttgct catgtgctca agacggatgc tgaccgccct tctgtgagcc tcgagctcct
     2461 caaccttagc caactgcatg acagcctggc cgacgttttt gagaccctcg gcagtcaagt
     2521 tggagctggt aatgtgcttg acgaacgagg cgagattgac gccagagtac tgcttggcaa
     2581 agccataggt agctatgtgt cttgacgtta gcagaagaat ctgtgaagcg gggctattta
     2641 tgatatgtaa cttacgcagc gagtggttaa caccagcaga gtaatcgcca acggactcag
     2701 gagtccaggc gccaatgaag acactaccgg cgttcatgac aagatcgaca gccttctcgg
     2761 cctccttgat ctggaggatc aagtgctcag gagcgtactt gttgctgagc tccatggcct
     2821 cctcgacggt cttgacctgc acggtgatcg agtgggcgat ggagccacgg acaatctgga
     2881 cgcgaggaag ctccgtagcc tgacggtgaa cctcgtcctc aatagcctga agatgctcct
     2941 cgtcgaggtc aatagcgatc aggatgacct gactgtcaac gccgtgctca gcctgggaca
     3001 ggagatccga ggcaacgaac gcggggttgg cgtccttgtc agcgatgacc agcacctcgg
     3061 acgggccagc gggcatgtca ataccaacgg cagcgttggt gtcgttgctg acgaacatct
     3121 tggcagcagt gacgaactgg ttaccggggc cgagaatctt gtcgaccttg gtgatgctct
     3181 cggtgccgta ggccatggca gctacggcct gggcaccgcc ggcaagcacg atggactcgg
     3241 cgccaacctt gtgagcgacg tagacaatct cgggagtgat ggttccgtcg gcgcggggag
     3301 gagaggcgaa cacaatcttg ttgcagccgg cgaccatggc gggaacaccc agcataaggg
     3361 cagtgctggg gagaacggcg gtaccgccgg ggatgtagca gccgacggcc tcgatgggac
     3421 gagagaaacg gctgcagaca acaccgggca tggtctcgac ctggaggggc ttctcctcct
     3481 tctgggcggc gtggaacttg cggatgttct cgaaggacac gtcgatggca gcaatggtct
     3541 cctcagggag ctgcataagc tccttgggga agggcgcctt caggacgggg ctagtaagag
     3601 aggtagcctt ctcgaacttg tgagtgtacg acagaacagc cttgtcgccg ttcttgcgga
     3661 cgtcctcgat gatggggaca atgatcttgt agatggcatc ggacgacttt tgcgcaggac
     3721 gcttgagagc agcatcgagc tcctcggtag agaccttgga ggcgtcgaaa cgtctcatgg
     3781 tgatcttctc aggggtggtc tcctgggcgg cctccttggt gacaggggcc gaagtggcag
     3841 cgagagcgga cgccgcaggc ttgatgccct ccttctcagc ccacttaccc ttagcatctc
     3901 cagtcctgcg cttgaccttc cagctcttgg cgtcaaggct cctttcgata tcggcaagag
     3961 taacgccggc agcaacggcc ctggtaagag caaagtagaa gagatcggca gcctcaaagg
     4021 cgatttcctg gggggtctga gcggtgcaga gctcctcagc ctcctccatg atcttggccc
     4081 ggactagctt ctcatcggag aagagacggg cagtgtagga gccctcgggg gcagactgtt
     4141 tcctcgaaat caaagtctgc tcgagcttgg gaaggccttt gagctgacca agcagccgga
     4201 ctgatcgagg tggcagaaac gacccttctg cttcacgaca aacttgagag catcgttatc
     4261 gcagtcaagc gagatgcgga cgagctcctg agtgtctccg gaagtagcac ccttgtacca
     4321 gagaccgcgc ttccggctct gatagacacc agtctgtgtc ctgagggcct cgttcacact
     4381 ctcggcactg ctgtagacca gacccagagc agtgtcgtgc tcatcaacga caacggtggg
     4441 gagaagacca tcgggacggt cagacttcca gacgctcgag aggatggtgg agatggcaag
     4501 cttcagctgc tcgagttcta tagtgtcacc taaatcgtat gtgtatgata cataaggtta
     4561 tgtattaatt gtagccgcgt tctaacgaca atatgtccat atggtgcact ctcagtacaa
     4621 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4681 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4741 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4801 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4861 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4921 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4981 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5041 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5101 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5161 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5221 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5281 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5341 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5401 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5461 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5521 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5581 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5641 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5701 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5761 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5821 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5881 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5941 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6001 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6061 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6721 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6781 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6841 ggttaacctg gcttatcgaa attaatacga ctcactatag ggagaccggc agatccaatg
     6901 cggatggatt cgctcactcg gtgccgcggc ggtgcgaagt tcctgccgtg gattggggga
     6961 ttgcacagac ggtggagtgc ttgctacagc taggtaccca gttgcacacc aaacggtgcc
     7021 cctgtgcccc tgcgtctagg tgtgtcctgt gagcaactgc gcccgcgcgc ggcaccggga
     7081 agctgcggga acctcaatgg atgattggcg ccagatcccc ggcatcacct cagcacatgg
     7141 gcagcccgag aaggaatcga gggtagaaga tattccaggg tttccgctag cacaacttcc
     7201 aagcgcctct accgctactc cacacaccca cgctgaatgg cacaaccact gaggcctagc
     7261 ggtcccagct ccccgtaacc acgcagcccc agcgaccgag gtcatcatga cactatgacg
     7321 acaatctggc gggagacttg ggaaaatcga tgacgggcac ctaccttggg aacaatagag
     7381 caagtccagt caacgcccta gggaaggggg aaaaaaaaaa aagagcccat tctcacaagg
     7441 agcaacactg tgctgctatc atagtacact acaagcaaag tcgggatgtt gctagaatcc
     7501 gtcggctacg tcggtagata cctcccatag gcagtaagtc taggtatcca atttatagtt
     7561 ccgcgtattt cttcgtcgaa cggggcacag caaagcctaa cagccgcagt gcctcgcaac
     7621 caacaccaag cagtaatctc atccatccat acagatcctc ccacaatatc tttacatgcg
     7681 cacttgcgct gtgttagcag tttaggttcc actaccaagg ctgacacgag cagcactgac
     7741 agcaaactgc tgcttggggc caagaaattg gcagtttgct gagcacctac atatcagtac
     7801 acatcaaccc tggtttaatt gttagcctgg tactgttaac cgtcatgaac gaggacgtgg
     7861 aggaagaacc acgctaacaa ttgaaccagg gttgatgtat acctctctta ccatatttgc
     7921 ttgcttccac gtccctatcc tcaagcgatt ttgttcttga ctttttgtat ttcaacgcaa
     7981 aggccattca ggtttctcat gttcaaccct ttggatggcg atgccgttcg tcccaggacc
     8041 tcgtgacctt tgaaagacga gaaatatatc ccaaagcaac ttctgcttat ggacaagcaa
     8101 ctgcatggcg gcccacaact catatctcgc gtaatcatcc aaacgctagg cccgagtact
     8161 ctactgttct cgagacacct accataatat gtacccaatc agacatgaca acagtcaacc
     8221 aaacataccc tgatccggcc aaaccctgac atccctagat gatggttagg cacacaaatg
     8281 gaaaacggga ctcgttggtt aggtgggaac gcttgtattg cagccagcca accgactctc
     8341 ctcgtccctc tcgagtcccg ttattgccgt ttgacccagc cgcccagccg ccaaacagct
     8401 gagggagcca atggtctaat acaccgtcca acggccccag tgatgtttca aacttgattg
     8461 acagcgaacg aaacccctga aacccgttat agatatgtat agatttgacg gggtagcttg
     8521 gccctaatac gactcactat agggcgaatt ggagctccac cgcggtggc
//
