LOCUS       sequence-139759-        7088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intermediate cloning vector, with MCS2 containing inducible unstable
            peptide driven by pGPD1 (UbiY 2xNLSs SZ3)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7088)
  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 7088)
  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: pDA186
FEATURES             Location/Qualifiers
     source          1..7088
                     /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     1040..1056
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1040..1056
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             1082..1309
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1445..1465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1481..1501
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      1742..1989
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1758..1776)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     2072..2090
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2128..2150)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2250..2269
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(2762..3856)
                     /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(3869..4273)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(4573..5028)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4710..4731)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4922..4941
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(5168..5185)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5339..5927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6098..6889,6890..6958))
                     /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     6721..6740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6959..7063)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtctaat acgactcact atagggcgaa ttggagctcg aagcccgaaa gagttatcgt
       61 tactccgatt attttgtaca gctgatggga ccttgccgtc ttcatttttt ttttttcacc
      121 tatagagccg ggcagagctg cccggcttaa ctaagggccg gaaaaaaaac ggaaaaaaga
      181 aagccaagcg tgtagacgta gtataacagt atatctgaca cgcacgtgat gaccacgtaa
      241 tcgcatcgcc cctcacctct catctctcac cgctgactca gcttcactaa aaaggaaaat
      301 atatactctt tcccaggcaa ggtgacagcg gtccccgtct cctccacaaa ggcctctcct
      361 ggggtttgag caagtctaag tttacgtagc ataaaaattc tcgcattgcg tcaaataata
      421 aaaaaagtaa ccccacttct acttctacat cggaaaaaca ttccattcac atatcgtctt
      481 tggcctatct tgttttgtcc tcggtagatc aggtcagtac aaacgcaaca cgaaagaaca
      541 aaaaaagaag aaaaacagaa ggccaagaca gggtcaatga gactgttgtc ctcctactgt
      601 ccctatgtct ctggccgatc acgcgccatt gtccctcaga aacaaatcaa acacccacac
      661 ccctggcacc caaagtcccc acccacacca ccaatacgta aacggggcgc cccctgcaag
      721 gccctcctgc gcgcggcctc ccgccttgct tctctcccct cccttttctt tttccagttt
      781 tccctatttt gtcccttttt ccgcacaaca agtatcagaa tgggttcatc aaatctatcc
      841 aacctaattc gcacgtagac tggcttggta ttggcagttt cgcagttata tatatactac
      901 catgagtgaa actgttacgt taccttaaat tctttctccc tttaattttc ttttatctta
      961 ctctcctaca taagacatca agaaacaatt gtatattgta caccccccct ccacaaacac
     1021 aaatattgat aatataaagt ctagaactag tggatccccc gggctgcagg aattcgatat
     1081 catgcagatt ttcgtcaaga ctttgaccgg taaaaccata acattggaag ttgaatcttc
     1141 cgataccatc gacaacgtta agtcgaaaat tcaagacaag gaaggtatcc ctccagatca
     1201 acaaagattg atctttgccg gtaagcagct agaagacggt agaacgctgt ctgattacaa
     1261 cattcagaag gagtccacct tacatcttgt gctaaggcta agaggtggtt accataaatc
     1321 aggtgcgtgg ctgcttccag ttagtttggt aaagagaaaa acgaccctag cacctaatac
     1381 tcaaacagct tcccccaggg ctttggccga ttctttaatg cagggaaagc ttatgagatc
     1441 cgaacccaag aagaagcgga aggtgggggc tggtgctgaa cccaagaaga agcggaaggt
     1501 ggggggatct agtgtcgacg gtggcagcaa tcaaacttcc ttatataaaa aagctggaag
     1561 tgccgcagca ccttttacaa tggaatttaa aagatctaac gaagttacca ctctggagaa
     1621 tgacgctgcg ttcatcgaaa atgaaaacgc ttacctggaa aaagaaatcg cgcgtctgcg
     1681 taaagaaaaa gcggcgctgc gcaaccgtct ggcgcacaaa aaatgataag ctagcctcga
     1741 gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc
     1801 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg
     1861 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg
     1921 tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct
     1981 ttaatttgcg gccggtaccc agcttttgtt ccctttagtg agggttaatg catgctaaga
     2041 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     2101 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     2161 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     2221 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     2281 ccatatcgac tacgtcgtaa ggccgtttct gacagagtaa aattcttgag ggaactttca
     2341 ccattatggg aaatggttca agaaggtatt gacttaaact ccatcaaatg gtcaggtcat
     2401 tgagtgtttt ttatttgttg tatttttttt tttttagaga aaatcctcca atatcaaatt
     2461 aggaatcgta gtttcatgat tttctgttac acctaacttt ttgtgtggtg ccctcctcct
     2521 tgtcaatatt aatgttaaag tgcaattctt tttccttatc acgttgagcc attagtatca
     2581 atttgcttac ctgtattcct ttactatcct cctttttctc cttcttgata aatgtatgta
     2641 gattgcgtat atagtttcgt ctaccctatg aacatattcc attttgtaat ttcgtgtcgt
     2701 ttctattatg aatttcattt ataaagttta tgtacaaata tcataaaaaa agagaatctt
     2761 tttaagcaag gattttctta acttcttcgg cgacagcatc accgacttcg gtggtactgt
     2821 tggaaccacc taaatcacca gttctgatac ctgcatccaa aaccttttta actgcatctt
     2881 caatggcctt accttcttca ggcaagttca atgacaattt caacatcatt gcagcagaca
     2941 agatagtggc gatagggtca accttattct ttggcaaatc tggagcagaa ccgtggcatg
     3001 gttcgtacaa accaaatgcg gtgttcttgt ctggcaaaga ggccaaggac gcagatggca
     3061 acaaacccaa ggaacctggg ataacggagg cttcatcgga gatgatatca ccaaacatgt
     3121 tgctggtgat tataatacca tttaggtggg ttgggttctt aactaggatc atggcggcag
     3181 aatcaatcaa ttgatgttga accttcaatg tagggaattc gttcttgatg gtttcctcca
     3241 cagtttttct ccataatctt gaagaggcca aaacattagc tttatccaag gaccaaatag
     3301 gcaatggtgg ctcatgttgt agggccatga aagcggccat tcttgtgatt ctttgcactt
     3361 ctggaacggt gtattgttca ctatcccaag cgacaccatc accatcgtct tcctttctct
     3421 taccaaagta aatacctccc actaattctc tgacaacaac gaagtcagta cctttagcaa
     3481 attgtggctt gattggagat aagtctaaaa gagagtcgga tgcaaagtta catggtctta
     3541 agttggcgta caattgaagt tctttacgga tttttagtaa accttgttca ggtctaacac
     3601 taccggtacc ccatttagga ccacccacag cacctaacaa aacggcatca accttcttgg
     3661 aggcttccag cgcctcatct ggaagtggga cacctgtagc atcgatagca gcaccaccaa
     3721 ttaaatgatt ttcgaaatcg aacttgacat tggaacgaac atcagaaata gctttaagaa
     3781 ccttaatggc ttcggctgtg atttcttgac caacgtggtc acctggcaaa acgacgatct
     3841 tcttaggggc agacataggg gcagacatta gaatggtata tccttgaaat atatatatat
     3901 attgctgaaa tgtaaaaggt aagaaaagtt agaaagtaag acgattgcta accacctatt
     3961 ggaaaaaaca ataggtcctt aaataatatt gtcaacttca agtattgtga tgcaagcatt
     4021 tagtcatgaa cgcttctcta ttctatatga aaagccggtt ccggcctctc acctttcctt
     4081 tttctcccaa tttttcagtt gaaaaaggta tatgcgtcag gcgacctctg aaattaacaa
     4141 aaaatttcca gtcatcgaat ttgattctgt gcgatagcgc ccctgtgtgt tctcgttatg
     4201 ttgaggaaaa aaataatggt tgctaagaga ttcgaactct tgcatcttac gatacctgag
     4261 tattcccaca gttaactgcg gtcaagatat ttcttgaatc aggcgcctta gaccgctcgg
     4321 ccaaacaacc aattacttgt tgagaaatag agtataatta tcctataaat ataacgtttt
     4381 tgaacacaca tgaacaagga agtacaggac aattgatttt gaagagaatg tggattttga
     4441 tgtaattgtt gggattccat ttttaataag gcaataatat taggtatgtg gatatactag
     4501 aagttctcct cgaccgtcga tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     4561 taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt
     4621 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4681 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4741 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4801 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4861 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4921 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4981 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg cgccattcgc
     5041 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5101 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5161 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5221 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5281 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5341 tccataggct cggcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5401 gaaacccgac aggactataa agataccagg cgttcccccc tggaagctcc ctcgtgcgct
     5461 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5521 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5581 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5641 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5701 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5761 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5821 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5881 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5941 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6001 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6061 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6121 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactg cccgtcgtgt
     6181 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6241 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6301 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6361 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6421 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6481 ggcgagttac atgatccccc atgttgtgaa aaaaagcggt tagctccttc ggtcctccga
     6541 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6601 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6661 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6721 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6781 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6841 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6901 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6961 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7021 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7081 tgccacct
//
