LOCUS       sequence-181187-        6748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  his7 integration plasmid with Z3EVpr:GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6748)
  AUTHORS   Ohira MJ, Hendrickson DG, Scott McIsaac R, Rhind N
  TITLE     An estradiol-inducible promoter enables fast, graduated control of 
            gene expression in fission yeast.
  JOURNAL   Yeast. 2017 Aug;34(8):323-334. doi: 10.1002/yea.3235. Epub 2017 Jun 
            8.
  PUBMED    28423198
REFERENCE   2  (bases 1 to 6748)
  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"; doi: "10.1002/yea.3235"; 
            journalName: "Yeast"; date: "2017-08"; volume: "34"; issue: "8"; 
            pages: "323-334"
COMMENT     Sequence Label: pFS462
FEATURES             Location/Qualifiers
     source          1..6748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    232..253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(268..273,274..291,292..298)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    306..322
                     /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     311..333
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     330..346
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     330..346
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        367..385
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     422..438
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     422..438
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     2951..2974
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2992..3705
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2992..3705
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3022..3050)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3645..3665
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3709..3729
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4146..4162)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4262..4278)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4263..4279)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4303..4322)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4304..4322)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4329..4345)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4487..4942
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4574..4593)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4784..4805
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4968..5072
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5073..5141,5142..5933)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGVS
                     LIKHW"
     primer_bind     complement(5291..5310)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6104..6692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6593..6612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      301 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      361 gctcgaaatt aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg
      421 ctctagaact agtggatccc ccggggatct cttttgagtc agaaaatctg acgcattgta
      481 acctggttac ctaaaggtga tttatgattg ttacttgggg cgatgtgaat ctttcgtttg
      541 ccaaacagaa aagtagttag agttttaggg gtatttgact tacaaatggc tcttttacca
      601 ttttttgatc ttacgaactt tgagagtgat gcttctgaag agttaggatg gcttaagtac
      661 gtcggcagag tgcaaacaag agtgtttcct cagcatttca aagataatct cgaaaaagtt
      721 cgcaaaattt cggaaacgat tgatgtgatc gttgacacga ctgctgaact tggaccagag
      781 gcctgcgtca accttttgaa cgctggtgca ctcgccatac tggttaatga agaaatgcta
      841 aacgagttgg cagacatctc tccgaatcgt ttggttttga agacagacac tactgatatt
      901 ggaaaaattg aaaaactttc acaagttgct ggttctattc aatggatcgg cagtgctgag
      961 aattatcccc ctgacttttt tgaaagagcc agcaagatca ttcacaaagc tgtaatgcct
     1021 gagggtggag gccgaacttt atatttggag ttccccgaac aaccatctat ggaggtactg
     1081 aaatcttttt ctgttcactc tgtcgttccc gttctgtcaa gttctttctt gacagtgaag
     1141 cctgccgaag agcccaagaa gttatcatta gctgatctca ttttgatatc tgccaatact
     1201 gatcgtgagg atggactatt ttctacttta gtcgtcaatg agctcggtat tgcacttggt
     1261 ctcgtgtatt ctagcaagga aagcgttgct gagtctttga agactggcac tggagtttat
     1321 caatcccgca aacgtggctt gtggtacaag ggcgcatctt ctggtgctgt tcaacactta
     1381 attcatattg acgtcgattg tgatgaagac tgcttgcgct ttgtcgtcta tcaaaccggt
     1441 aaaggttttt gtcatcttga caccttgcat tgctttggac aagcttcagg actttgtcag
     1501 ttagaaaaga ctttaatcga taggaagaat aatgctcctg agggatctta tactgctcgt
     1561 ttgttctccg acccaaagtt attgcgtgca aaaattatgg aggaagctga agagttgtgt
     1621 gatgctacca ccaaagagaa tgttatttgg gaaatggctg atttgatgta ctttgccata
     1681 acccgctgcg tcggttcggg tgttagcctt aatgacatta gtcgtcatct tgatttgaaa
     1741 catcgtaaag ttacaagaag aaagggagat gctaaagttg catggcaaga gaaacttaaa
     1801 gacaaaggag gagtggcaaa cacgtcttat actgcctgaa ttttttaaat ttctactctt
     1861 cattcgcaga tgatagcagt cggtctaagg tgtagtctac taaaaccttg ctaatgcgga
     1921 aatttttact ttgtgtaatt atttaataat gaaaaaaaat tcatatttgc tgtttgtact
     1981 aatattattt gttttatatt gtttaatttg agcgctggat agataataac tcagacttca
     2041 ttagccactg gctggtattt gttgctcgac ttattcctac caaatatcga ccttgtcgct
     2101 gtgtatacct tctgtaatac actagaatta gttgaactct ctgtttcaac atcatctgct
     2161 tcactgaata taaggctctt aggctttttc catctcctga gaagtaagct acttgcaaga
     2221 acactaagag aagaaaacat catagcagca ctagcccaca taggatttaa gtagattccc
     2281 catggaagga agttatattg aattttcaaa aattcttact ttttttttgg atggacgcaa
     2341 agaagtttaa taatcatatt acatggcatt accaccatat acatatccat atacatatcc
     2401 atatctaatc ttacttatat gttgtggaaa tgtaaagagc cccattatct tagcctaaaa
     2461 aaaccttctc tttggaactt tcagtaatac gcttaactgc tcattgctat attgaagtgc
     2521 ggccgcgtgg gcgtgcgtgg gcgggcgtgg gcgtgcgtgg gcgggcgtgg gcgtgcgtgg
     2581 gcgtctagac cgtgcgtcct cgtcttcacc ggtcgcgttc ctgaaacgca gatgtgcctc
     2641 gcgccgcact gctccgaaca ataaagattc tacaatacta gcttttatgg ttatgaagag
     2701 gaaaaattgg cagtaacctg gccccacaaa ccttcaaatt aacgaatcaa attaacaacc
     2761 ataggatgat aatgcgatta gttttttagc cttatttctg gggtaattaa tcagcgaagc
     2821 gatgattttt gatctattaa cagatatata aatggaaaag ctgcataacc actttaacta
     2881 atactttcaa cattttcagt ttgtattact tcttattcaa atgtcataaa agtatcaaca
     2941 aaaaattgtt aatatacctc tatactttaa cgtcaaggag aaaaaactat aatgagtaaa
     3001 ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat
     3061 gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc
     3121 cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact
     3181 ctcacttatg gtgttcaatg cttttcaaga tacccagatc acatgaaaca gcatgacttt
     3241 ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac
     3301 gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc
     3361 gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac
     3421 aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt
     3481 aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa
     3541 caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca
     3601 caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt
     3661 gtaacagctg ctgggattac acatggcatg gatgaactat acaaagggcc taagaagaag
     3721 cgtaaggttt aatagggttt aatgtagaat aaattcatat gatttgattt gacctaattt
     3781 ttattgatta attgtgtcgc atatttatta tcgttaaaaa tgcaattgac aaaagcgtaa
     3841 attttaaggc tctaatgttt tttcttttgg tatgttatag tcaagcaaag atgtattaag
     3901 ttgcaaaagc atggaattga tttacgacat ccgttcaact ttatgaccga gttgtcgctg
     3961 tgtacttact ctttttttac tatgaaaagt ttcaaacata atatcattaa ttatttcttt
     4021 ttaaacgaag taaagcccat tgcaatagga atcatgacaa agttatatat gcaagcccac
     4081 acaaggttca ttttaattct tttaacgata actctagagt cgacctgcag gcatgcaagc
     4141 ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     4201 aatcgccttg cagcacatcc ccctttcgcc aggggctgca ggaattcgat atcaagctta
     4261 tcgataccgt cgacctcgag ggggggcccg gtacccaatt cgccctatag tgagtcgtat
     4321 tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     4381 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     4441 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     4501 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4561 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4621 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4681 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4741 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4801 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4861 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4921 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4981 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5041 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5101 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5161 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5221 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5281 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5341 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5401 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5461 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5521 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5581 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5641 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5701 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5761 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5821 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5881 aacgaaatag acagatcgct gagataggtg tctcactgat taagcattgg taactgtcag
     5941 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6001 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6061 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6121 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6181 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6241 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6301 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6361 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6421 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6481 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6541 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6601 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6661 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6721 tcctggcctt ttgctggcct tttgctca
//
