LOCUS       sequence-139758-        7281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intermediate cloning vector, with MCS2 containing inducible stable 
            peptide driven by pSTL1 (Venus 2xNLSs SZ1)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7281)
  AUTHORS   Aymoz D, Wosika V, Durandau E, Pelet S
  TITLE     Real-time quantification of protein expression at the single-cell 
            level via dynamic protein synthesis translocation reporters.
  JOURNAL   Nat Commun. 2016 Apr 21;7:11304. doi: 10.1038/ncomms11304.
  PUBMED    27098003
REFERENCE   2  (bases 1 to 7281)
  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: "Nat 
            Commun."; date: "2016-04-21"; volume: "7:11304. doi"; pages: " 
            10.1038/ncomms11304"
COMMENT     Sequence Label: pDA171
FEATURES             Location/Qualifiers
     source          1..7281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..26
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     840..856
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     840..856
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             900..920
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             936..956
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             978..1688
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     ICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     protein_bind    1716..1740
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1935..2182
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1951..1969)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     2265..2283
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2321..2343)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2443..2462
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(2955..4049)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4062..4466)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(4766..5221)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4903..4924)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5115..5134
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(5361..5378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5532..6120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6291..7082,7083..7151))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6914..6933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7152..7256)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtctaat acgactcact atagggcgaa ttggagctcg cagaacacca ctactgatac
       61 tcattgccaa ggctaggagg caccatccgt ttcattttct ttgaggtaag ccaatcatga
      121 aatagtatac acatccataa cggacgtacg gacgaaataa gtgccgttgt cccactattc
      181 caccgcattt ggcccatttg gctcactttg actcaacttg cgtcatttta actgatatga
      241 agggtccgac tttgtccttt ttcggccacc gcatacccca cggcgatgcc tccgctacct
      301 gcatttgagt agcatctccg tttcgcgggg tattcggcgc tacgtcgcct gttcgagcgg
      361 ctctgttcgt tgcatgaaac taaaataagc ggaaagtgtc cagccatcca ctacgtcaga
      421 aagaaataat ggttgtacac tgtttctcgg ctatataccg tttttggttg gttaatcctc
      481 gccaggtgca gctattgcgc ttggctgctt cgcgatagta gtaatctgag aaagtgcaga
      541 tcccggtaag ggaaacactt ttggttcacc tttgataggg ctttcattgg ggcattcgta
      601 acaaaaagga agtagataga gaaattgaga aagcttaagt gagatgtttt agcttcaatt
      661 ttgtcccctt caacgctgct tggccttaga gggtcagaat tgcagttcag gagtagtcac
      721 actcatagta tataaacaag ccctttattg attttgaata attattttgt atacgtgttc
      781 tagcatacaa gttagaataa ataaaaaata gaaaaataga acatagaaag ttttagacct
      841 ctagaactag tggatccccc gggctgcagg aattcgatat caagcttatg agatccgaac
      901 ccaagaagaa gcggaaggtg ggggctggtg ctgaacccaa gaagaagcgg aaggtggggg
      961 gatctagtag atctgcttct aaaggtgaag aattattcac tggtgttgtc ccaattttgg
     1021 ttgaattaga tggtgatgtt aatggtcaca aattttctgt ctccggtgaa ggtgaaggtg
     1081 atgctactta cggtaaattg accttaaaat tgatttgtac tactggtaaa ttgccagttc
     1141 catggccaac cttagtcact actttaggtt atggtttgca atgttttgct agatacccag
     1201 atcatatgaa acaacatgac tttttcaagt ctgccatgcc agaaggttat gttcaagaaa
     1261 gaactatttt tttcaaagat gacggtaact acaagaccag agctgaagtc aagtttgaag
     1321 gtgatacctt agttaataga atcgaattaa aaggtattga ttttaaagaa gatggtaaca
     1381 ttttaggtca caaattggaa tacaactata actctcacaa tgtttacatc actgctgaca
     1441 aacaaaagaa tggtatcaaa gctaacttca aaattagaca caacattgaa gatggtggtg
     1501 ttcaattagc tgaccattat caacaaaata ctccaattgg tgatggtcca gtcttgttac
     1561 cagacaacca ttacttatcc tatcaatctg ccttatccaa agatccaaac gaaaagagag
     1621 accacatggt cttgttagaa tttgttactg ctgctggtat tacccatggt atggatgaat
     1681 tgtacaaagg atccgtcgac ggtgggtcga atcaaacaag tttgtacaaa aaagcaggct
     1741 ccgcggccgc ccccttcacc atggaattca aaagatctaa cctggttgcg cagctcgaaa
     1801 acgaagttgc gtctctggaa aatgagaacg aaaccctgaa gaaaaagaac ctgcacaaaa
     1861 aagacctgat cgcgtacctg gagaaagaaa tcgcgaatct gcgtaagaaa atcgaagaat
     1921 aagctagcct cgagtcatgt aattagttat gtcacgctta cattcacgcc ctccccccac
     1981 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
     2041 ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     2101 gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg
     2161 acgctcgaag gctttaattt gcggccggta cccagctttt gttcccttta gtgagggtta
     2221 atgcatgcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     2281 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     2341 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     2401 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     2461 tactgagagt gcaccatatc gactacgtcg taaggccgtt tctgacagag taaaattctt
     2521 gagggaactt tcaccattat gggaaatggt tcaagaaggt attgacttaa actccatcaa
     2581 atggtcaggt cattgagtgt tttttatttg ttgtattttt ttttttttag agaaaatcct
     2641 ccaatatcaa attaggaatc gtagtttcat gattttctgt tacacctaac tttttgtgtg
     2701 gtgccctcct ccttgtcaat attaatgtta aagtgcaatt ctttttcctt atcacgttga
     2761 gccattagta tcaatttgct tacctgtatt cctttactat cctccttttt ctccttcttg
     2821 ataaatgtat gtagattgcg tatatagttt cgtctaccct atgaacatat tccattttgt
     2881 aatttcgtgt cgtttctatt atgaatttca tttataaagt ttatgtacaa atatcataaa
     2941 aaaagagaat ctttttaagc aaggattttc ttaacttctt cggcgacagc atcaccgact
     3001 tcggtggtac tgttggaacc acctaaatca ccagttctga tacctgcatc caaaaccttt
     3061 ttaactgcat cttcaatggc cttaccttct tcaggcaagt tcaatgacaa tttcaacatc
     3121 attgcagcag acaagatagt ggcgataggg tcaaccttat tctttggcaa atctggagca
     3181 gaaccgtggc atggttcgta caaaccaaat gcggtgttct tgtctggcaa agaggccaag
     3241 gacgcagatg gcaacaaacc caaggaacct gggataacgg aggcttcatc ggagatgata
     3301 tcaccaaaca tgttgctggt gattataata ccatttaggt gggttgggtt cttaactagg
     3361 atcatggcgg cagaatcaat caattgatgt tgaaccttca atgtagggaa ttcgttcttg
     3421 atggtttcct ccacagtttt tctccataat cttgaagagg ccaaaacatt agctttatcc
     3481 aaggaccaaa taggcaatgg tggctcatgt tgtagggcca tgaaagcggc cattcttgtg
     3541 attctttgca cttctggaac ggtgtattgt tcactatccc aagcgacacc atcaccatcg
     3601 tcttcctttc tcttaccaaa gtaaatacct cccactaatt ctctgacaac aacgaagtca
     3661 gtacctttag caaattgtgg cttgattgga gataagtcta aaagagagtc ggatgcaaag
     3721 ttacatggtc ttaagttggc gtacaattga agttctttac ggatttttag taaaccttgt
     3781 tcaggtctaa cactaccggt accccattta ggaccaccca cagcacctaa caaaacggca
     3841 tcaaccttct tggaggcttc cagcgcctca tctggaagtg ggacacctgt agcatcgata
     3901 gcagcaccac caattaaatg attttcgaaa tcgaacttga cattggaacg aacatcagaa
     3961 atagctttaa gaaccttaat ggcttcggct gtgatttctt gaccaacgtg gtcacctggc
     4021 aaaacgacga tcttcttagg ggcagacata ggggcagaca ttagaatggt atatccttga
     4081 aatatatata tatattgctg aaatgtaaaa ggtaagaaaa gttagaaagt aagacgattg
     4141 ctaaccacct attggaaaaa acaataggtc cttaaataat attgtcaact tcaagtattg
     4201 tgatgcaagc atttagtcat gaacgcttct ctattctata tgaaaagccg gttccggcct
     4261 ctcacctttc ctttttctcc caatttttca gttgaaaaag gtatatgcgt caggcgacct
     4321 ctgaaattaa caaaaaattt ccagtcatcg aatttgattc tgtgcgatag cgcccctgtg
     4381 tgttctcgtt atgttgagga aaaaaataat ggttgctaag agattcgaac tcttgcatct
     4441 tacgatacct gagtattccc acagttaact gcggtcaaga tatttcttga atcaggcgcc
     4501 ttagaccgct cggccaaaca accaattact tgttgagaaa tagagtataa ttatcctata
     4561 aatataacgt ttttgaacac acatgaacaa ggaagtacag gacaattgat tttgaagaga
     4621 atgtggattt tgatgtaatt gttgggattc catttttaat aaggcaataa tattaggtat
     4681 gtggatatac tagaagttct cctcgaccgt cgatatgcgg tgtgaaatac cgcacagatg
     4741 cgtaaggaga aaataccgca tcaggaaatt gtaaacgtta atattttgtt aaaattcgcg
     4801 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4861 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4921 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4981 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5041 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5101 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5161 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5221 tcgcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     5281 tcgctattac gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5341 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5401 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5461 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5521 ttgctggcgt ttttccatag gctcggcccc cctgacgagc atcacaaaaa tcgacgctca
     5581 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgttccc ccctggaagc
     5641 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5701 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     5761 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5821 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5881 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5941 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6001 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6061 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6121 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6181 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6241 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6301 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6361 ctgcccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6421 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6481 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6541 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6601 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6661 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gaaaaaaagc ggttagctcc
     6721 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6781 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6841 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6901 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6961 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7021 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7081 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7141 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7201 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7261 tttccccgaa aagtgccacc t
//
