LOCUS       sequence-54340-e        7438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7438)
  AUTHORS   Zilio N, Wehrkamp-Richter S, Boddy MN
  TITLE     A new versatile system for rapid control of gene expression in the 
            fission yeast Schizosaccharomyces pombe.
  JOURNAL   Yeast. 2012 Aug 29. doi: 10.1002/yea.2920.
  PUBMED    22968950
REFERENCE   2  (bases 1 to 7438)
  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: "Yeast. 2012
            Aug 29. doi: 10.1002/yea.2920."
COMMENT     Sequence Label: pUG6SP-tTA'
FEATURES             Location/Qualifiers
     source          1..7438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     45..64
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        45..63
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(117..150)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            complement(213..1569)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      complement(213..410)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(416..1225)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1139..1158
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1226..1569)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     protein_bind    complement(1624..1657)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        1763..2142
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2143..2346
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2297..2317
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2343..2367
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2447..3067,3068..3454)
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the reverse tetracycline repressor 
                     rTetR with the C-terminal activation domain of herpes 
                     simplex virus VP16"
                     /label=rtTA
                     /note="In the Tet-On(R) system, rtTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLKGESWQDFLRNNAKSFRCALLSHRNGAKVHSDTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(2520..2539)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(5137..5155)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5138..5155)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5236..5255)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5355..5377
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5415..5433)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5501..5605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5606..5674,5675..6466)
                     /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(5824..5843)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6637..7225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7126..7145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7379..7396
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgttggccga ttcattaatg caggttaacc tggcttatcg aaattaatac gactcactat
       61 agggagaccg gcagatccgc ggccgcatag gccactagtg gatctgatat cacctaataa
      121 cttcgtatag catacattat acgaagttat attaagggtt ctcgagagct cgttttcgac
      181 actggatggc ggcgttagta tcgaatcgac agcagtatag cgaccagcat tcacatacga
      241 ttgacgcatg atattacttt ctgcgcactt aacttcgcat ctgggcagat gatgtcgagg
      301 cgaaaaaaaa tataaatcac gctaacattt gattaaaata gaacaactac aatataaaaa
      361 aactatacaa atgacaagtt cttgaaaaca agaatctttt tattgtcagt actgattaga
      421 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
      481 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
      541 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
      601 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
      661 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
      721 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
      781 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
      841 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
      901 ctaatacctg gaatgctgtt ttgccgggga tcgcagtggt gagtaaccat gcatcatcag
      961 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     1021 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     1081 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     1141 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg
     1201 aaacgtgagt cttttcctta cccatggttg tttatgttcg gatgtgatgt gagaactgta
     1261 tcctagcaag attttaaaag gaagtatatg aaagaagaac ctcagtggca aatcctaacc
     1321 ttttatattt ctctacaggg gcgcggcgtg gggacaattc aacgcgtctg tgaggggagc
     1381 gtttccctgc tcgcaggtct gcagcgagga gccgtaattt ttgcttcgcg ccgtgcggcc
     1441 atcaaaatgt atggatgcaa atgattatac atggggatgt atgggctaaa tgtacgggcg
     1501 acagtcacat catgcccctg agctgcgcac gtcaagactg tcaaggaggg tattctgggc
     1561 ctccatgtcg ctggccgggt gacccggcgg ggacgaggca agctaaacag atctctagac
     1621 ctaataactt cgtatagcat acattatacg aagttatatt aagggttgtc gacgctcgag
     1681 gagcttggcc cattgcatac gttgtatcca tatcataata tgtacattta tattggctca
     1741 tgtccaacat taccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt
     1801 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     1861 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     1921 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     1981 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2041 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2101 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2161 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2221 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2281 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2341 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
     2401 catagaagac accgggaccg atccagcctc cgcggcccca attcatatgt ctagattaga
     2461 taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt
     2521 aacaacccgt aaactcgccc agaagcttgg tgtagagcag cctacactgt attggcatgt
     2581 aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac
     2641 tcacttttgc cctttaaaag gggaaagctg gcaagatttt ttacgcaata acgctaaaag
     2701 ttttagatgt gctttactaa gtcatcgcaa tggagcaaaa gtacattcag atacacggcc
     2761 tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt
     2821 ttcactagag aacgcgttat atgcactcag cgctgtgggg cattttactt taggttgcgt
     2881 attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag
     2941 tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc
     3001 cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag
     3061 tgggtccgcg tacagccgcg cgcgtacgaa aaacaattac gggtctacca tcgagggcct
     3121 gctcgatctc ccggacgacg acgcccccga agaggcgggg ctggcggctc cgcgcctgtc
     3181 ctttctcccc gcgggacaca cgcgcagact gtcgacggcc cccccgaccg atgtcagcct
     3241 gggggacgag ctccacttag acggcgagga cgtggcgatg gcgcatgccg acgcgctaga
     3301 cgatttcgat ctggacatgt tgggggacgg ggattccccg ggtccgggat ttacccccca
     3361 cgactccgcc ccctacggcg ctctggatat ggccgacttc gagtttgagc agatgtttac
     3421 cgatgccctt ggaattgacg agtacggtgg gtagggggcg cgaggatcgt aataagcttc
     3481 gacctgcagc gtacgaagct tcagctgggg taaaaggaat gtctcccttg ccagtactgc
     3541 tagggttttt ctttcaaact atggaagccc attcaagctg catattacga ttttgttttt
     3601 cgcttttaga aagtggttta gatgagataa tagaaaaatt cttgatctcc gacaacgagt
     3661 acttttattt tttttgctaa tcactttact caatattagc tcgaaatcgt agaaacgtag
     3721 acgggtgcgg gataccgagt ggtgtagtta agaattttta taaaccacgt ggcccaaaaa
     3781 tatgaaccca aaacgtttat acatgagtat actttaagaa ggctataccc cttcgtgtta
     3841 gatgtagttt tagctaccca acccgagtct atgagcttga cttcagatgt agaaggcatt
     3901 aaatcgtttt gaatattaat taaaaaacga tgaaaattaa atatttaaaa gcaatcatac
     3961 gctgaaaatt tagtgctgtg gctaatcctt caacatggaa atgccataaa agtgactttg
     4021 acaaaaaaaa aagtatatac aggtagtaaa ctcatctact tcattgactt tgtttacagc
     4081 atgtggaagg aggaatattt attgctaaat cgtagtttaa cattcaataa gtaatactat
     4141 tgaaattcga caagattggc cgcatggatg aaaaagaggc attttgcttt gggagaatta
     4201 gttcaaatta gaactgaaaa aaaaaacttt acgaggcaaa aatgtcggat tgagatcgta
     4261 aaagttcgct cgtcgtcttt tgctttgtga ttgttttcat ggatacatct tgctggatat
     4321 ttaaatttta gtactatgta taagatattc tataaatgtt ttatcaccca aacctgttag
     4381 cgccttctta attctattca atctggcttt tgctctgaga ctacttcttg gactttcact
     4441 acttgttagt tatacggaat ttgtgtaatt agaagtgaaa taatcctttc tattagtaat
     4501 gcgagctcct tgatttatac atgaaagcat tcttttgaaa caatattttg ttttctattt
     4561 gatttcgcgt tgaatagtcc caatgaccct gatgagacaa aagcacttat cataaaattt
     4621 attatctaag ataatctcgt taaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaatttggca
     4681 agaagccctt ctctttcgtt aaaacttgtt tcgctttatg caatactgta agtgtaagct
     4741 tgtaatatag cgaatctgtg aactcattgt tcatttcatg gtagttgttg atttcacaag
     4801 catgctgagt tatcaatata gtaacacaat tggttttatt tcagctcgct tgaatatagt
     4861 gtcaagagat aattgaataa acaaaaaaaa taactacacg ttttttagat acattttatt
     4921 tacaatgtat tggttcttta aagaatgggt ggtaaataaa aaaaatatta caaggtcgaa
     4981 aaaaattcat cccttttcat tagttctata attaaatgct aaacccatga atattctttt
     5041 actgctaatc aattaacgat tttggtattt aaaagctatt gctgataagt attctgacgt
     5101 tctgtatcaa tattaccatg cagctggcgg ccgcgttcta tagtgtcacc taaatcgtat
     5161 gtgtatgata cataaggtta tgtattaatt gtagccgcgt tctaacgaca atatgtccat
     5221 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     5281 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5341 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5401 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5461 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5521 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5581 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5641 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5701 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5761 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5821 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5881 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5941 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6001 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6061 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6121 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6181 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6241 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6301 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6361 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6421 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6481 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6541 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6601 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6661 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6721 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6781 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6841 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6901 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6961 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7021 gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7081 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7141 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7201 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7261 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7321 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7381 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcg
//
