LOCUS       sequence-140105-        6900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intermediate cloning vector, with MCS2 containing inducible unstable
            peptide driven by pSTL1 (UbiY 2xNLSs SZ1)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6900)
  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 6900)
  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: pDA125
FEATURES             Location/Qualifiers
     source          1..6900
                     /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             882..1109
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1245..1265
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1281..1301
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      1554..1801
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1570..1588)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     1884..1902
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1940..1962)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2062..2081
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(2574..3668)
                     /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(3681..4085)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(4385..4840)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4522..4543)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4734..4753
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(4980..4997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5151..5739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5910..6701,6702..6770))
                     /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     6533..6552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6771..6875)
                     /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 catgcagatt ttcgtcaaga
      901 ctttgaccgg taaaaccata acattggaag ttgaatcttc cgataccatc gacaacgtta
      961 agtcgaaaat tcaagacaag gaaggtatcc ctccagatca acaaagattg atctttgccg
     1021 gtaagcagct agaagacggt agaacgctgt ctgattacaa cattcagaag gagtccacct
     1081 tacatcttgt gctaaggcta agaggtggtt accataaatc aggtgcgtgg ctgcttccag
     1141 ttagtttggt aaagagaaaa acgaccctag cacctaatac tcaaacagct tcccccaggg
     1201 ctttggccga ttctttaatg cagggaaagc ttatgagatc cgaacccaag aagaagcgga
     1261 aggtgggggc tggtgctgaa cccaagaaga agcggaaggt ggggggatct agtgtcgacg
     1321 gtggcagcaa tcaaacttcc ttatataaaa aagctggaag tgccgcagca ccttttacaa
     1381 tggaatttaa aagatctaac ctggttgcgc agctcgaaaa cgaagttgcg tctctggaaa
     1441 atgagaacga aaccctgaag aaaaagaacc tgcacaaaaa agacctgatc gcgtacctgg
     1501 agaaagaaat cgcgaatctg cgtaagaaaa tcgaagaata agctagcctc gagtcatgta
     1561 attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga
     1621 aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt
     1681 aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg
     1741 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg
     1801 cggccggtac ccagcttttg ttccctttag tgagggttaa tgcatgctaa gaaaccatta
     1861 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     1921 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     1981 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     2041 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatcg
     2101 actacgtcgt aaggccgttt ctgacagagt aaaattcttg agggaacttt caccattatg
     2161 ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc attgagtgtt
     2221 ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatcaaa ttaggaatcg
     2281 tagtttcatg attttctgtt acacctaact ttttgtgtgg tgccctcctc cttgtcaata
     2341 ttaatgttaa agtgcaattc tttttcctta tcacgttgag ccattagtat caatttgctt
     2401 acctgtattc ctttactatc ctcctttttc tccttcttga taaatgtatg tagattgcgt
     2461 atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc gtttctatta
     2521 tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc tttttaagca
     2581 aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact gttggaacca
     2641 cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc ttcaatggcc
     2701 ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga caagatagtg
     2761 gcgatagggt caaccttatt ctttggcaaa tctggagcag aaccgtggca tggttcgtac
     2821 aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg caacaaaccc
     2881 aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat gttgctggtg
     2941 attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc agaatcaatc
     3001 aattgatgtt gaaccttcaa tgtagggaat tcgttcttga tggtttcctc cacagttttt
     3061 ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat aggcaatggt
     3121 ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac ttctggaacg
     3181 gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct cttaccaaag
     3241 taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc aaattgtggc
     3301 ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct taagttggcg
     3361 tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac actaccggta
     3421 ccccatttag gaccacccac agcacctaac aaaacggcat caaccttctt ggaggcttcc
     3481 agcgcctcat ctggaagtgg gacacctgta gcatcgatag cagcaccacc aattaaatga
     3541 ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag aaccttaatg
     3601 gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat cttcttaggg
     3661 gcagacatag gggcagacat tagaatggta tatccttgaa atatatatat atattgctga
     3721 aatgtaaaag gtaagaaaag ttagaaagta agacgattgc taaccaccta ttggaaaaaa
     3781 caataggtcc ttaaataata ttgtcaactt caagtattgt gatgcaagca tttagtcatg
     3841 aacgcttctc tattctatat gaaaagccgg ttccggcctc tcacctttcc tttttctccc
     3901 aatttttcag ttgaaaaagg tatatgcgtc aggcgacctc tgaaattaac aaaaaatttc
     3961 cagtcatcga atttgattct gtgcgatagc gcccctgtgt gttctcgtta tgttgaggaa
     4021 aaaaataatg gttgctaaga gattcgaact cttgcatctt acgatacctg agtattccca
     4081 cagttaactg cggtcaagat atttcttgaa tcaggcgcct tagaccgctc ggccaaacaa
     4141 ccaattactt gttgagaaat agagtataat tatcctataa atataacgtt tttgaacaca
     4201 catgaacaag gaagtacagg acaattgatt ttgaagagaa tgtggatttt gatgtaattg
     4261 ttgggattcc atttttaata aggcaataat attaggtatg tggatatact agaagttctc
     4321 ctcgaccgtc gatatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     4381 caggaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4441 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4501 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4561 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4621 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4681 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4741 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4801 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cgcgccattc gccattcagg
     4861 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcat
     4921 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4981 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5041 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5101 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5161 ctcggccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5221 acaggactat aaagatacca ggcgttcccc cctggaagct ccctcgtgcg ctctcctgtt
     5281 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5341 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5401 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5461 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5521 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5581 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5641 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5701 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5761 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5821 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5881 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5941 tcagcgatct gtctatttcg ttcatccata gttgcctgac tgcccgtcgt gtagataact
     6001 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6061 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6121 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6181 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6241 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6301 acatgatccc ccatgttgtg aaaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6361 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6421 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6481 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6541 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6601 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6661 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6721 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6781 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6841 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
//
