LOCUS       sequence-251698-        7761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing synthetic TetR-GFP protein in N. crassa (from the csr-1 
            locus).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7761)
  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 7761)
  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: pTSN7B
FEATURES             Location/Qualifiers
     source          1..7761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     4125..4141
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4175..4191)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4175..4191)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             join(5251..5253,5254..5256,5257..5970)
                     /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(5296..5317)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5557..5576)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5904..5925
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5991..6007)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6036..6054)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7117..7136)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7118..7136)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7146..7163)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7146..7162)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7155..7177)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(7303..7758)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7440..7461)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7652..7671
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct gggtaccggg cccaacacct
     2221 ccgtcgccat aaactcctcc agaatagcca ccgtaggtag agctgctacc accgccacca
     2281 ctgctgctac cgccatacga agtgttgccg aaaccgccat aacgtccgct gctcccgctg
     2341 gagaagctca ttccacccga aatgcccgtg aacttgccct tgtttgcgcg ggccttcttg
     2401 cgctctgctc ggatacgctc gacatcgctg aggagctcgg cgagttcctt ggcgcggttg
     2461 cggacgttga taccctggtc tttgccgttc tggtcgataa aatggaactg gcgcagcatc
     2521 ttgaggaggg taagatgcga ccgagcatcg tccaccactc gttcggaacc gtgcttaatc
     2581 aggtactcta gtagttgaag ggccttgtag atctggcgcc attcttcggc ggacttttct
     2641 gtgaaacggc ggtaaatcat gggcatgatt tcgttgagtg tttggctaag aacaaggagg
     2701 gaaacgcaag ttagccgcgt tcgtatacaa gcgagcagtt ctcgagaacg tactagttga
     2761 aggtgccatc ggcgatctct tgcatcaaag tcgacgatgc accccaaggt tcattgttgg
     2821 tggcctcgcg cacctgggtt cgctgtcagc tcccgttcca tggtctcttg tctgggatca
     2881 agtatatggc gaagggcgca agagaatggt gccagtgtgg cgcgtggata cgggcatacc
     2941 ttggactcca tctcggtata gttcatgaca gctggttgga gccattgtca gcaccctgaa
     3001 cacgaaatca tgagggagtc ttggttctgg actaggtggt cacctaccat tttgaacctt
     3061 tcgtacgcca gccttgatgt cgtagagtgt caggttggac acggtatcct tgaggctatt
     3121 gaaatccatt gcgtaaacgt gtctatcgcc ggagttgctg ggaattgttc tgatgggcta
     3181 aggtaattct tctctagatc caatctgagc ctcgcggtga tgattgccag acgggcggga
     3241 tgctgtggcg acttggtggt ttttgttcgg gctgatcgct ttcagggcga cgtttcacgg
     3301 tcttgtcggg atcagatgga cattgaggtg gcctagcgtc agggtgccgc tgagcgctga
     3361 cctcaactga cgccgtccaa ggccgacccc tgcgcttgac ccaacaccct gacaccaaga
     3421 agccagtgga acccaggagg tctgccaaga ctttgaaagc ttggcacgag cgtctcagct
     3481 ctttgaagcc tgagttttca ggcaagaatt ctgcttgtaa ttgccgggcc ggccgacaac
     3541 gcggggcggc gggaggggac gctcggccgc cggagctcgc ttccatgcat tttggatgga
     3601 gtaaaaattg ttaaacaccc gacccgactc cgttctccct ctcgtctcag gtctttgttg
     3661 tgcatccatc gccgtgtagc agatttccat cgttcgtttg ttcggacttt accttgtact
     3721 tttagcttca ttatcatcct acaatcatcc aggatttcat cttccacaac cggtacgtta
     3781 tcatttttac ttttgttgcc atgttcttct tgagcctttt ctgtctagag gagctctccc
     3841 tgccggctgg gagcttgccc aaagctccaa gctctatctg tggcagagct ctgagagctt
     3901 ccattcccgg gatttcttgg catcagattc aattcgaatg tccttttttc catccacgct
     3961 cgagatgagg ggtgaggttt gttagtttga caataccacc accgcctttt ggtccctacc
     4021 ccagacatgg ccatcaaggt cgttaatgcc gtccatccct cttgttcggc ttggggtatc
     4081 gtccatggct ccgcaatgtc gtcaatggct gcagggcccc ccctcgaggt cgacggtatc
     4141 gataagcttg atatcgaatt cctgcagccc gggggatcca ctagttctag agcggccgca
     4201 gaagatgata ttgaaggagc actttttggg cttggctgga gctagtggag gtcaacaatg
     4261 aatgcctatt ttggtttagt cgtccaggcg gtgagcacaa aatttgtgtc gtttgacaag
     4321 atggttcatt taggcaactg gtcagatcag ccccacttgt agcagtagcg gcggcgctcg
     4381 aagtgtgact cttattagca gacaggaacg aggacattat tatcatctgc tgcttggtgc
     4441 acgataactt ggtgcgtttg tcaagcaagg taagtgaacg acccggtcat accttcttaa
     4501 gttcgccctt cctcccttta tttcagattc aatctgactt acctattcta ccgaagctct
     4561 agaatggcca tgtcccgcct cgacaagtcc aaggtcatca actccgctct cgagctcctc
     4621 aacgaggtcg gtatcgaggg tctcaccacc cgcaagctcg ctcagaagct cggtgtcgag
     4681 cagcctaccc tctactggca cgtcaagaac aagcgcgctc tcctcgacgc tctcgccatc
     4741 gagatgctcg accgccacca cacccacttc agccctctcg agggtgagtc ctggcaggac
     4801 ttcctccgca acaacgccaa gtccttccgc aacgctctgc tctcccaccg cgacggtgct
     4861 aaggtccacc tcggtacccg ccctaccgag aagcagtacg agaccctcga gaaccagctc
     4921 gctttcctca cccagcaggg cttctccctc gagaacgccc tctacgctct ctccgctgtc
     4981 ggtcacttca ccctcggctc cgtcctcgag gaccaggagc accaggtcgc caaggaggaa
     5041 cgcgagaccc ctaccaccga ctccatgcct cctctcctcc gccaggccat cgagctcttc
     5101 gaccaccagg gtgccgagcc tgccttcctc cacggcctcg agtccctcat ccgcggtttc
     5161 gaggtccagc tcaccgctct cctccagatc gtctccggta gcggcttaat cagcgctcct
     5221 aggcgcaaga agcgcaagac cggtattaac atggtgagca agggcgagga gctgttcacc
     5281 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     5341 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     5401 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcaccta cggcgtgcag
     5461 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     5521 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     5581 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     5641 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     5701 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     5761 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     5821 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     5881 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     5941 actcacggca tggacgagct gtacaagtaa gaattcgata tcaagcttat cgataccgtc
     6001 gacctcgagg gggggcccgg taccagcttt tgttcccttt agtgagggtt aattagggcg
     6061 cctcctaggc ttaagccgcg gtggatttcc tgcgctgcac acccaccctt cttcactgca
     6121 tcgtcatcac tgtcgctttg atggttgatg tgattacgca aactgatggt tgatgagatc
     6181 acgtaaactg atgtcttttc ctagacagtt tgcaggtcat gtgaatgtta tcgccaacac
     6241 attgaatgct aaacagatta cctccaacgc ctagatgaaa ccaaattacc cttgccttct
     6301 ttttttttac gccgttaatg cagttgtgat agagaccatg caacgtcttc taaacggaga
     6361 cacaaacaca ttcgagtcga tcagatcgaa atgacttgtt cctgtacctc gtcaccatcc
     6421 agccaccgtc tgaaatagac gcagttgctg gggctgacag gtccgccttc ctcgcacata
     6481 tcatagacag ggcaccgccc acaaggcgcc tccataaggc cacttgcact cggctcaact
     6541 cccataaatg aggtaacaat gcctcctttg gttggatcat ggtcgaggcg tcgttgattg
     6601 cacgtcctcg gatcctgttt ggggatgcgc gtcacacggt accccttgcg cccattgacc
     6661 ttgacaggct ctactaagcc atcgtagtat agcacatcca ccaactgctg gatatccgcc
     6721 tcgctcaacg tcgtattatt cgtgatgccg gcgtggtgga tgaactttgc aatgtccgac
     6781 acagtcgggt acgacttgta accggccggg agaggcagca attgcggctt gccgggcctg
     6841 gtcttcttgg ccggtgccgg tgccgccgtg tcgtcgttgg aaatctcgcc cgcgctgcgc
     6901 ttcgtcccct tggaaatatc ctccgagtgg ggaatggtcc cctgaaccac gcccttcttg
     6961 ggattctttt cacgctgctg tgctcctcct ccttgtgctc ctactcctgg tactccagca
     7021 ccgccgccgt gggaagtata tgtcgatttc tgcttgataa agtcaaacac aatttcctgt
     7081 agctcgttga tgaaggccgt gtcgagctcc aattcgccct atagtgagtc gtattacgcg
     7141 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     7201 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     7261 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggaaattgta agcgttaata
     7321 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     7381 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     7441 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     7501 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     7561 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     7621 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     7681 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     7741 cgccgctaca gggcgcgtca g
//
