LOCUS       sequence-443809-        6746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB2 ZWF1 tet-on overexpression cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6746)
  AUTHORS   Matthias Steiger
  TITLE     Regulation of the Glucose-6-phosphate dehydrogenase encoding gene 
            gsdA and its impact on growth and citric acid production in 
            Aspergillus niger
  JOURNAL   PLOS One
REFERENCE   2  (bases 1 to 6746)
  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: "PLOS One"
COMMENT     Sequence Label: pSF18
FEATURES             Location/Qualifiers
     source          1..6746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..73
                     /label=L4440
                     /note="L4440 vector, forward primer"
     intron          812..927
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     CDS             join(945..1562,1563..1568,1569..1604,1605..1607,1608..1643,
                     1644..1646,1647..1682,1683..1691)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     protein_bind    join(2039..2057,2058..2080,2081..2099,2100..2122,2123..2141,
                     2142..2164,2165..2183,2184..2206,2207..2225,2226..2248,
                     2249..2267,2268..2290,2291..2309)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     intron          2379..2494
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     terminator      4033..4806
                     /gene="trpC"
                     /label=trpC terminator
                     /note="transcription terminator from the Aspergillus 
                     nidulans trpC gene (Punt et al., 1987)"
     promoter        4869..4973
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4974..5042,5043..5834)
                     /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(5192..5211)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6060..6648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6549..6568
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
       61 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
      121 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
      181 caacactagt cgcaaaaaac cccgcccctg acagggcggg gttttttcgc ggtctccgat
      241 cggagtcgat aagcttcgga gaatatggag cttcatcgaa tcaccggcag taagcgaagg
      301 agaatgtgaa gccaggggtg tatagccgtc ggcgaaatag catgccatta acctaggtac
      361 agaagtccaa ttgcttccga tctggtaaaa gattcacgag atagtacctt ctccgaagta
      421 ggtagagcga gtacccggcg cgtaagctcc ctaattggcc catccggcat ctgtagggcg
      481 tccaaatatc gtgcctctcc tgctttgccc ggtgtatgaa accggaaagg ccgctcagga
      541 gctggccagc ggcgcagacc gggaacacaa gctggcagtc gacccatccg gtgctctgca
      601 ctcgacctgc tgaggtccct cagtccctgg taggcagctt tgccccgtct gtccgcccgg
      661 tgtgtcggcg gggttgacaa ggtcgttgcg tcagtccaac atttgttgcc atattttcct
      721 gctctcccca ccagctgctc ttttcttttc tctttctttt cccatcttca gtatattcat
      781 cttcccatcc aagaaccttt atttccccta agtaagtact ttgctacatc catactccat
      841 ccttcccatc ccttattcct ttgaaccttt cagttcgagc tttcccactt catcgcagct
      901 tgactaacag ctaccccgct tgagcagaca tcaccgaatt caccatgtct agactggaca
      961 agagcaaagt cataaacggc gctctggaat tactcaatgg agtcggtatc gaaggcctga
     1021 cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac tggcacgtga
     1081 agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg catcataccc
     1141 acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac gccaagtcat
     1201 tccgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc acccgcccaa
     1261 cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct
     1321 ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg ggctgcgtat
     1381 tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc accgattcta
     1441 tgcccccact tctgagacaa gcaattgagc tgttcgaccg gcagggagcc gaacctgcct
     1501 tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag tgcgaaagcg
     1561 gcgggccggc cgacgccctt gacgattttg acttagacat gctcccagcc gatgcccttg
     1621 acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac cttgacatgc
     1681 tccccgggta actaagtaag gatccactag tacagcagaa gaatctctct ccgctgttgc
     1741 ttcagtgtct gccatgcatt aacttcatcc tactgtccta cccgcagtac ccattcacat
     1801 ttgcggcaga taccaggttg ttttctattc cctcggttct tcagttcttc agatattaca
     1861 ttactcgaga aattggcggc gatgcaggag tttggttggt tttaattgtt tcagtctcct
     1921 tgacctgatt tgtatgacat gcaatgtttc gacggatgac tatctcggtg aatatacgtc
     1981 atgaatcatg cggccgcgcg tatcacgagg ccctttcgac ttcactcgag tttaccactc
     2041 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
     2101 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta
     2161 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat
     2221 agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg
     2281 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagctcccc atcttcagta
     2341 tattcatctt cccatccaag aacctttatt tcccctaagt aagtactttg ctacatccat
     2401 actccatcct tcccatccct tattcctttg aacctttcag ttcgagcttt cccacttcat
     2461 cgcagcttga ctaacagcta ccccgcttga gcagacatca ccgtttaaac accatgaccg
     2521 atacgaaagc cgtagaattt gtgggccaca cagccattgt agtctttgga gcttcagggg
     2581 acctggctaa gaagaaaact ttccctgccc tcttcggact ttaccgtgag ggatacctgt
     2641 ccaacaaggt gaagattatt ggctatgcta gatcaaagct ggatgacaag gagttcaagg
     2701 atagaattgt gggctatttc aagacaaaga acaagggcga cgaggacaaa gttcaagaat
     2761 tcttaaagtt gtgctcatat atttcagctc cttatgacaa accagatggg tatgaaaagt
     2821 tgaatgaaac tattaacgaa ttcgaaaagg aaaacaacgt cgaacagtct cacaggttgt
     2881 tctacttagc tttgccccct tctgttttca tacctgttgc tacggaggtc aagaagtatg
     2941 ttcatccagg ttctaaaggg attgctcgga ttatcgtgga aaaacctttc gggcacgact
     3001 tgcagtcagc agaagagctt ttgaatgctt tgaagccgat ctggaaagaa gaggaattgt
     3061 ttagaatcga ccactatcta ggtaaggaga tggttaagaa tttgttggcc ttccgttttg
     3121 gaaacgcatt catcaatgct tcttgggaca acagacatat cagctgtatc caaatctcgt
     3181 tcaaggagcc ttttggaaca gaaggtcgtg gtggctattt tgactcaatt ggtataataa
     3241 gagacgtcat tcagaaccac ttgcttcaag tgttaaccct cttaaccatg gaaagacccg
     3301 tctctaatga ccctgaggct gttagagatg aaaaggttcg cattctgaag tcaatttctg
     3361 agctagattt gaacgacgtt ttggtgggtc aatacggcaa atctgaggat ggaaagaagc
     3421 cagcttatgt ggatgatgaa actgttaagc caggttctaa atgtgtcaca tttgcagcca
     3481 ttggcttgca catcaacaca gaaaggtggg aaggtgtccc aatcatttta agagctggta
     3541 aggctttgaa cgaaggtaaa gttgagatta gagtgcaata caaacagtct actggatttc
     3601 tcaatgatat tcagcgaaat gaattggtca tccgtgtgca gcctaacgaa gccatgtaca
     3661 tgaaactgaa ctccaaagtc ccaggtgttt cccaaaagac tactgtcact gagctagacc
     3721 tcacttacaa agaccgttac gaaaactttt acattccaga ggcatatgaa tcacttatca
     3781 gagatgctat gaagggagat cactctaatt ttgtcagaga tgacgagttg atacaaagtt
     3841 ggaagatttt cactccttta ctgtatcact tggagggccc tgatgcaccg gctccagaaa
     3901 tctatcccta cggatccaga ggtccagctt cattgaccaa attcttgcaa gatcatgatt
     3961 acttctttga atcacgcgac aattaccaat ggccagtgac aagacccgat gtgctgcaca
     4021 agatgtaagc ttgatccact taacgttact gaaatcatca aacagcttga cgaatctgga
     4081 tataagatcg ttggtgtcga tgtcagctcc ggagttgaga caaatggtgt tcaggatctc
     4141 gataagatac gttcatttgt ccaagcagca aagagtgcct tctagtgatt taatagctcc
     4201 atgtcaacaa gaataaaacg cgtttcgggt ttacctcttc cagatacagc tcatctgcaa
     4261 tgcattaatg cattggacct cgcaacccta gtacgccctt caggctccgg cgaagcagaa
     4321 gaatagctta gcagagtcta ttttcatttt cgggagacga gatcaagcag atcaacggtc
     4381 gtcaagagtc ctacgagact gaggaatccg ctcttggctc cacgcgacta tatatttgtc
     4441 tctaattgta ctttgacatg ctcctcttct ttactctgat agcttgacta tgaaaattcc
     4501 gtcaccagcc cctgggttcg caaagataat tgcactgttt cttccttgaa ctctcaagcc
     4561 tacaggacac acattcatcg taggtataaa cctcgaaaat cattcctact aagatgggta
     4621 tacaatagta accatgcatg gttgcctagt gaatgctccg taacacccaa tacgccggcc
     4681 gaaacttttt tacaactctc ctatgagtcg tttacccaga atgcacaggt acacttgttt
     4741 agaggtaatc cttctttcta gaagtcctcg tgtactgtgt aagcgcccac tccacatctc
     4801 cactcgctcc ggggagacca ccggtgccgc tacagggcgc gtcaggtggc acttttcggg
     4861 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4921 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4981 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5041 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5101 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5161 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5221 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5281 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5341 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5401 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5461 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5521 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5581 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5641 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggc tctcgcggta
     5701 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5761 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5821 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5881 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaacaata
     5941 aaactgtctg cttacataaa cagtaataca aggggtgtag atctcatgac caaaatccct
     6001 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6061 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6121 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6181 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6241 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6301 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6361 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6421 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6481 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6541 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6601 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6661 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6721 ttatcccctg attctgtgga taaccg
//
