LOCUS       pTRE3G_NlucP            4019 bp ds-DNA     circular     26-OCT-2020
DEFINITION  Tet/Dox-inducible expression of EGFP in mammalian cells (control).
ACCESSION   .
VERSION     .
KEYWORDS    pTetON-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4100)
  AUTHORS   Harterink M, da Silva ME, Will L, Turan J, Ibrahim A, Lang AE, van
            Battum EY, Pasterkamp RJ, Kapitein LC, Kudryashov D, Barres BA,
            Hoogenraad CC, Zuchero JB
  TITLE     DeActs: genetically encoded tools for perturbing the actin
            cytoskeleton in single cells.
  JOURNAL   Nat Methods. 2017 Apr 10. doi: 10.1038/nmeth.4257.
  PUBMED    28394337
REFERENCE   2  (bases 1 to 4100)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     gene            435..947
                     /gene="NlucP"
                     /locus_tag="NlucP"
                     /label="NlucP"
                     /ApEinfo_label="NlucP"
                     /ApEinfo_fwdcolor="#ff797d"
                     /ApEinfo_revcolor="#ff102f"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     intron          1212..1277
                     /note="SV40 (simian virus 40) small t antigen intron"
                     /locus_tag="small t intron"
                     /label="small t intron"
                     /ApEinfo_label="small t intron"
                     /ApEinfo_fwdcolor="#eb606c"
                     /ApEinfo_revcolor="#eb606c"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             1407..1427
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /translation="PKKKRKV"
                     /locus_tag="SV40 NLS"
                     /label="SV40 NLS"
                     /ApEinfo_label="SV40 NLS"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     polyA_signal    1852..1973
                     /note="SV40 polyadenylation signal"
                     /locus_tag="SV40 poly(A) signal"
                     /label="SV40 poly(A) signal"
                     /ApEinfo_label="SV40 poly(A) signal"
                     /ApEinfo_fwdcolor="#ff3eee"
                     /ApEinfo_revcolor="#ff3eee"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(1889..1908)
                     /note="SV40 polyA, reverse primer"
                     /locus_tag="SV40pA-R"
                     /label="SV40pA-R"
                     /ApEinfo_label="SV40pA-R"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     1943..1962
                     /note="SV40 polyA terminator, reverse primer"
                     /locus_tag="EBV-rev"
                     /label="EBV-rev"
                     /ApEinfo_label="EBV-rev"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(2059..2076)
                     /note="L4440 vector, forward primer"
                     /locus_tag="L4440"
                     /label="L4440"
                     /ApEinfo_label="L4440"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     rep_origin      complement(2230..2818)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /locus_tag="ori"
                     /label="ori"
                     /ApEinfo_label="ori"
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     primer_bind     complement(2310..2329)
                     /note="pBR322 origin, forward primer"
                     /locus_tag="pBR322ori-F"
                     /label="pBR322ori-F"
                     /ApEinfo_label="pBR322ori-F"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     CDS             complement(2989..3849)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     I
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     S
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     W
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     A
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     S LIKHW"
                     /locus_tag="AmpR"
                     /label="AmpR"
                     /ApEinfo_label="AmpR"
                     /ApEinfo_fwdcolor="#e9d024"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     3612..3631
                     /note="Ampicillin resistance gene, reverse primer"
                     /locus_tag="Amp-R"
                     /label="Amp-R"
                     /ApEinfo_label="Amp-R"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     CDS             948..1070
                     /locus_tag="PEST sequence"
                     /label="PEST sequence"
                     /ApEinfo_label="PEST sequence"
                     /ApEinfo_fwdcolor="#e624cc"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        complement(3850..3954)
                     /gene="bla"
                     /locus_tag="AmpR promoter"
                     /label="AmpR promoter"
                     /ApEinfo_label="AmpR promoter"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /ApEinfo_hidden
     primer_bind     3..21
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
                     /locus_tag="pBRforEco"
                     /label="pBRforEco"
                     /ApEinfo_label="pBRforEco"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     332..356
                     /note="Human CMV promoter, forward primer"
                     /locus_tag="LNCX"
                     /label="LNCX"
                     /ApEinfo_label="LNCX"
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    269..287
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /locus_tag="tet operator"
                     /label="tet operator"
                     /ApEinfo_label="tet operator"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    233..251
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /locus_tag="tet operator(1)"
                     /label="tet operator(1)"
                     /ApEinfo_label="tet operator"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    197..215
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /locus_tag="tet operator(2)"
                     /label="tet operator(2)"
                     /ApEinfo_label="tet operator"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    161..179
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /locus_tag="tet operator(3)"
                     /label="tet operator(3)"
                     /ApEinfo_label="tet operator"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    125..143
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /locus_tag="tet operator(4)"
                     /label="tet operator(4)"
                     /ApEinfo_label="tet operator"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    89..107
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /locus_tag="tet operator(5)"
                     /label="tet operator(5)"
                     /ApEinfo_label="tet operator"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    53..71
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /locus_tag="tet operator(6)"
                     /label="tet operator(6)"
                     /ApEinfo_label="tet operator"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        45..423
                     /note="3rd-generation Tet-responsive promoter that can be
                     activated by binding of Tet-On(R) 3G"
                     /locus_tag="TRE3G promoter"
                     /label="TRE3G promoter"
                     /ApEinfo_label="TRE3G promoter"
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
ORIGIN
        1 aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt cctcgagttt actccctatc
       61 agtgatagag aacgtatgaa gagtttactc cctatcagtg atagagaacg tatgcagact
      121 ttactcccta tcagtgatag agaacgtata aggagtttac tccctatcag tgatagagaa
      181 cgtatgacca gtttactccc tatcagtgat agagaacgta tctacagttt actccctatc
      241 agtgatagag aacgtatatc cagtttactc cctatcagtg atagagaacg tataagcttt
      301 aggcgtgtac ggtgggcgcc tataaaagca gagctcgttt agtgaaccgt cagatcgcct
      361 ggagcaattc cacaacactt ttgtcttata ccaactttcc gtaccacttc ctaccctcgt
      421 aaagtcgagc caccatggtc ttcacactcg aagatttcgt tggggactgg cgacagacag
      481 ccggctacaa cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc
      541 tcggggtgtc cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga
      601 tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg
      661 aaaaaatttt taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact
      721 atggcacact ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt
      781 atgaaggcat cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg
      841 gcaacaaaat tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa
      901 ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat tctggcgaat tctcacggct
      961 ttccgcctga ggttgaagag caagccgccg gtacattgcc tatgtcctgc gcacaagaaa
     1021 gcggtatgga ccggcaccca gccgcttgtg cttcagctcg catcaacgtc TAAtaagatc
     1081 caatgtaact gtattcagcg atgacgaaat tcttagctat tgtaatactc tagaggatct
     1141 ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct acagagattt
     1201 aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa
     1261 ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag tggtggaatg
     1321 cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat gatgaggcta
     1381 ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa gaccccaagg
     1441 actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat agaactcttg
     1501 cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag aaaattatgg
     1561 aaaaatattc tgtaaccttt ataagtaggc ataacagtta taatcataac atactgtttt
     1621 ttcttactcc acacaggcat agagtgtctg ctattaataa ctatgctcaa aaattgtgta
     1681 cctttagctt tttaatttgt aaaggggtta ataaggaata tttgatgtat agtgccttga
     1741 ctagagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     1801 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     1861 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1921 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     1981 tgcggctcta gagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2041 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2101 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2161 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2221 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2281 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2341 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2401 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2461 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2521 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2581 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2641 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2701 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2761 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2821 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2881 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2941 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3001 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3061 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3121 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3181 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3241 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3301 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3361 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3421 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3481 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3541 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3601 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3661 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3721 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3781 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3841 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3901 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3961 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctata
//
