LOCUS       sequence-255818-        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"
     CDS             join(1060..1062,1063..1065,1066..1779)
                     /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(1105..1126)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1366..1385)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1713..1734
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1800..1816)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1845..1863)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2926..2945)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2927..2945)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2955..2972)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2955..2971)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2964..2986)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3112..3567)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3249..3270)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3461..3480
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3594..3698
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3699..3767,3768..4559)
                     /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(3917..3936)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4730..5318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5219..5238
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5472..5489
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5606..5627
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5642..5647,5648..5665,5666..5672)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5680..5696
                     /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     5685..5707
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5704..5720
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5704..5720
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5741..5759
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     7695..7711
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7745..7761)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7745..7761)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 gcggccgcag aagatgatat tgaaggagca ctttttgggc ttggctggag ctagtggagg
       61 tcaacaatga atgcctattt tggtttagtc gtccaggcgg tgagcacaaa atttgtgtcg
      121 tttgacaaga tggttcattt aggcaactgg tcagatcagc cccacttgta gcagtagcgg
      181 cggcgctcga agtgtgactc ttattagcag acaggaacga ggacattatt atcatctgct
      241 gcttggtgca cgataacttg gtgcgtttgt caagcaaggt aagtgaacga cccggtcata
      301 ccttcttaag ttcgcccttc ctccctttat ttcagattca atctgactta cctattctac
      361 cgaagctcta gaatggccat gtcccgcctc gacaagtcca aggtcatcaa ctccgctctc
      421 gagctcctca acgaggtcgg tatcgagggt ctcaccaccc gcaagctcgc tcagaagctc
      481 ggtgtcgagc agcctaccct ctactggcac gtcaagaaca agcgcgctct cctcgacgct
      541 ctcgccatcg agatgctcga ccgccaccac acccacttca gccctctcga gggtgagtcc
      601 tggcaggact tcctccgcaa caacgccaag tccttccgca acgctctgct ctcccaccgc
      661 gacggtgcta aggtccacct cggtacccgc cctaccgaga agcagtacga gaccctcgag
      721 aaccagctcg ctttcctcac ccagcagggc ttctccctcg agaacgccct ctacgctctc
      781 tccgctgtcg gtcacttcac cctcggctcc gtcctcgagg accaggagca ccaggtcgcc
      841 aaggaggaac gcgagacccc taccaccgac tccatgcctc ctctcctccg ccaggccatc
      901 gagctcttcg accaccaggg tgccgagcct gccttcctcc acggcctcga gtccctcatc
      961 cgcggtttcg aggtccagct caccgctctc ctccagatcg tctccggtag cggcttaatc
     1021 agcgctccta ggcgcaagaa gcgcaagacc ggtattaaca tggtgagcaa gggcgaggag
     1081 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1141 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1201 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcacctac
     1261 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1321 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1381 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1441 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1501 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1561 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1621 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     1681 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1741 gccgggatca ctcacggcat ggacgagctg tacaagtaag aattcgatat caagcttatc
     1801 gataccgtcg acctcgaggg ggggcccggt accagctttt gttcccttta gtgagggtta
     1861 attagggcgc ctcctaggct taagccgcgg tggatttcct gcgctgcaca cccacccttc
     1921 ttcactgcat cgtcatcact gtcgctttga tggttgatgt gattacgcaa actgatggtt
     1981 gatgagatca cgtaaactga tgtcttttcc tagacagttt gcaggtcatg tgaatgttat
     2041 cgccaacaca ttgaatgcta aacagattac ctccaacgcc tagatgaaac caaattaccc
     2101 ttgccttctt tttttttacg ccgttaatgc agttgtgata gagaccatgc aacgtcttct
     2161 aaacggagac acaaacacat tcgagtcgat cagatcgaaa tgacttgttc ctgtacctcg
     2221 tcaccatcca gccaccgtct gaaatagacg cagttgctgg ggctgacagg tccgccttcc
     2281 tcgcacatat catagacagg gcaccgccca caaggcgcct ccataaggcc acttgcactc
     2341 ggctcaactc ccataaatga ggtaacaatg cctcctttgg ttggatcatg gtcgaggcgt
     2401 cgttgattgc acgtcctcgg atcctgtttg gggatgcgcg tcacacggta ccccttgcgc
     2461 ccattgacct tgacaggctc tactaagcca tcgtagtata gcacatccac caactgctgg
     2521 atatccgcct cgctcaacgt cgtattattc gtgatgccgg cgtggtggat gaactttgca
     2581 atgtccgaca cagtcgggta cgacttgtaa ccggccggga gaggcagcaa ttgcggcttg
     2641 ccgggcctgg tcttcttggc cggtgccggt gccgccgtgt cgtcgttgga aatctcgccc
     2701 gcgctgcgct tcgtcccctt ggaaatatcc tccgagtggg gaatggtccc ctgaaccacg
     2761 cccttcttgg gattcttttc acgctgctgt gctcctcctc cttgtgctcc tactcctggt
     2821 actccagcac cgccgccgtg ggaagtatat gtcgatttct gcttgataaa gtcaaacaca
     2881 atttcctgta gctcgttgat gaaggccgtg tcgagctcca attcgcccta tagtgagtcg
     2941 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3001 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3061 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaaattgtaa
     3121 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3181 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3241 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3301 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3361 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3421 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3481 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3541 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3601 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3661 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3721 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3781 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3841 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3901 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3961 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4021 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4081 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4141 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4201 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4261 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4321 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4381 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4441 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4501 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4561 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4621 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4681 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4741 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4801 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4861 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4921 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4981 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5041 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5101 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5161 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5221 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5281 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5341 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5401 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     5461 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     5521 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     5581 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     5641 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     5701 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
     5761 aacaaaagct gggtaccggg cccaacacct ccgtcgccat aaactcctcc agaatagcca
     5821 ccgtaggtag agctgctacc accgccacca ctgctgctac cgccatacga agtgttgccg
     5881 aaaccgccat aacgtccgct gctcccgctg gagaagctca ttccacccga aatgcccgtg
     5941 aacttgccct tgtttgcgcg ggccttcttg cgctctgctc ggatacgctc gacatcgctg
     6001 aggagctcgg cgagttcctt ggcgcggttg cggacgttga taccctggtc tttgccgttc
     6061 tggtcgataa aatggaactg gcgcagcatc ttgaggaggg taagatgcga ccgagcatcg
     6121 tccaccactc gttcggaacc gtgcttaatc aggtactcta gtagttgaag ggccttgtag
     6181 atctggcgcc attcttcggc ggacttttct gtgaaacggc ggtaaatcat gggcatgatt
     6241 tcgttgagtg tttggctaag aacaaggagg gaaacgcaag ttagccgcgt tcgtatacaa
     6301 gcgagcagtt ctcgagaacg tactagttga aggtgccatc ggcgatctct tgcatcaaag
     6361 tcgacgatgc accccaaggt tcattgttgg tggcctcgcg cacctgggtt cgctgtcagc
     6421 tcccgttcca tggtctcttg tctgggatca agtatatggc gaagggcgca agagaatggt
     6481 gccagtgtgg cgcgtggata cgggcatacc ttggactcca tctcggtata gttcatgaca
     6541 gctggttgga gccattgtca gcaccctgaa cacgaaatca tgagggagtc ttggttctgg
     6601 actaggtggt cacctaccat tttgaacctt tcgtacgcca gccttgatgt cgtagagtgt
     6661 caggttggac acggtatcct tgaggctatt gaaatccatt gcgtaaacgt gtctatcgcc
     6721 ggagttgctg ggaattgttc tgatgggcta aggtaattct tctctagatc caatctgagc
     6781 ctcgcggtga tgattgccag acgggcggga tgctgtggcg acttggtggt ttttgttcgg
     6841 gctgatcgct ttcagggcga cgtttcacgg tcttgtcggg atcagatgga cattgaggtg
     6901 gcctagcgtc agggtgccgc tgagcgctga cctcaactga cgccgtccaa ggccgacccc
     6961 tgcgcttgac ccaacaccct gacaccaaga agccagtgga acccaggagg tctgccaaga
     7021 ctttgaaagc ttggcacgag cgtctcagct ctttgaagcc tgagttttca ggcaagaatt
     7081 ctgcttgtaa ttgccgggcc ggccgacaac gcggggcggc gggaggggac gctcggccgc
     7141 cggagctcgc ttccatgcat tttggatgga gtaaaaattg ttaaacaccc gacccgactc
     7201 cgttctccct ctcgtctcag gtctttgttg tgcatccatc gccgtgtagc agatttccat
     7261 cgttcgtttg ttcggacttt accttgtact tttagcttca ttatcatcct acaatcatcc
     7321 aggatttcat cttccacaac cggtacgtta tcatttttac ttttgttgcc atgttcttct
     7381 tgagcctttt ctgtctagag gagctctccc tgccggctgg gagcttgccc aaagctccaa
     7441 gctctatctg tggcagagct ctgagagctt ccattcccgg gatttcttgg catcagattc
     7501 aattcgaatg tccttttttc catccacgct cgagatgagg ggtgaggttt gttagtttga
     7561 caataccacc accgcctttt ggtccctacc ccagacatgg ccatcaaggt cgttaatgcc
     7621 gtccatccct cttgttcggc ttggggtatc gtccatggct ccgcaatgtc gtcaatggct
     7681 gcagggcccc ccctcgaggt cgacggtatc gataagcttg atatcgaatt cctgcagccc
     7741 gggggatcca ctagttctag a
//
