LOCUS       sequence-254190-        2828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Position 3 module- 3SV40NLS  (3x NLS from SV40).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2828)
  AUTHORS   Goulev Y, Matifas A, Heyer V, Reina-San-Martin B, Charvin G
  TITLE     COSPLAY: An expandable toolbox for combinatorial and swift 
            generation of expression plasmids in yeast.
  JOURNAL   PLoS One. 2019 Aug 28;14(8):e0220694. doi: 
            10.1371/journal.pone.0220694. eCollection 2019.
  PUBMED    31461461
REFERENCE   2  (bases 1 to 2828)
  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: "PLoS One.";
            date: "2019-08-28"; volume: "14(8):e0220694. doi"; pages: " 
            10.1371/journal.pone.0220694. eCollection 2019"
COMMENT     Sequence Label: pMV45
FEATURES             Location/Qualifiers
     source          1..2828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..137
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     129..146
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     130..146
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             220..240
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             244..264
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             268..288
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(358..374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(358..374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(371..393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    382..398
                     /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)."
     promoter        complement(join(406..412,413..430,431..436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    451..472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(589..606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(760..1348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(840..859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1519..2310,2311..2379))
                     /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     2142..2161
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2380..2484)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2552..2570
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2608..2630)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2730..2749
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
       61 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      121 cacgacgttg taaaacgacg gccagtgaat tcgagctcgg tacctcgcga atgcatctag
      181 atatcggatc ccggtctcgg aacaatggtg cacgtggacc caaaaaagaa gagaaaggta
      241 gatccaaaaa agaagagaaa ggtagatcca aaaaagaaga gaaaggtaca cgtgtccggc
      301 gcaagagacc gggcccgtcg actgcagagg cctgcatgca agcttggcgt aatcatggtc
      361 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
      421 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
      481 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
      541 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
      601 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
      661 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
      721 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      781 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      841 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      901 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      961 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1021 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1081 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1141 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1201 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1261 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1321 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1381 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1441 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1501 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1561 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1621 gggcttacca tctggcccca gtgctgcaat gataccgcga gatccacgct caccggctcc
     1681 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     1741 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     1801 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     1861 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     1921 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1981 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2041 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2101 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2161 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2221 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2281 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2341 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     2401 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     2461 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     2521 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     2581 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     2641 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     2701 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     2761 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     2821 ttcgccat
//
