LOCUS       sequence-96649-e        5758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of NLS-mCherry (control plasmid). Confers 
            resistance to Puromycin. Expression is inducible in T-REx cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5758)
  AUTHORS   Britton S, Dernoncourt E, Delteil C, Froment C, Schiltz O, Salles B,
            Frit P, Calsou P
  TITLE     DNA damage triggers SAF-A and RNA biogenesis factors exclusion from 
            chromatin coupled to R-loops removal.
  JOURNAL   Nucleic Acids Res. 2014 Oct 1;42(14):9047-62. doi: 
            10.1093/nar/gku601. Epub 2014 Jul 16.
  PUBMED    25030905
REFERENCE   2  (bases 1 to 5758)
  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"; doi: 
            "10.1093/nar/gku601"; journalName: "Nucleic Acids Res"; date: 
            "2014-10-1- 1"; volume: "42"; issue: "14"; pages: "9047-62"
COMMENT     Sequence Label: pICE-NLS-mCherry
FEATURES             Location/Qualifiers
     source          1..5758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             24..44
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      54..63
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             60..770
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(205..223)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     473..492
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(838..855)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    844..1068
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1114..1542
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1201..1220)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1411..1432
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1551..1571)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1556..1885
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1736..1871
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1798..1817
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1937..2536
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1937..1956)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2433..2453
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2666..2787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2703..2722)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2757..2776
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2837..2853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2837..2853)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2850..2872)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2861..2877
                     /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(2885..2891,2892..2909,2910..2915))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2930..2951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3068..3085)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3239..3824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3319..3338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3995..4786,4787..4855))
                     /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     4618..4637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4856..4960)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5032..5411
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5412..5615
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5566..5586
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    5617..5635
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5638..5656
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     5666..5690
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 agcttgccac catgacgcgt ggccccaaga aaaagcggaa agtgggaccg gtcgccacca
       61 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
      121 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
      181 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
      241 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
      301 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
      361 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
      421 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
      481 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
      541 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
      601 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
      661 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
      721 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa agcggccgcg
      781 aattcggatc cggtaccctc gagacgcgtc tgcagtctag ataaacccgc tgatcagcct
      841 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
      901 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
      961 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     1021 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     1081 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     1141 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     1201 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     1261 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     1321 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     1381 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     1441 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     1501 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     1561 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     1621 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     1681 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     1741 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     1801 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     1861 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     1921 tgatcagcac gtgaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     1981 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     2041 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     2101 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     2161 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     2221 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     2281 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     2341 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     2401 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     2461 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     2521 caagcccggt gcctgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     2581 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     2641 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     2701 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2761 tttgtccaaa ctcatcaatg tatcttatca tgtctggcta gccgtcgacc tctagctaga
     2821 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2881 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2941 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3001 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3061 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3121 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3181 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3241 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3301 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3361 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3421 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3481 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3541 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3601 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3661 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3721 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     3781 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3841 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3901 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3961 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4021 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4081 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4141 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4201 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4261 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4321 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4381 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4441 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4501 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4561 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4621 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4681 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4741 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4801 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4861 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4921 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4981 cacctgacgt cgacggatcg ggagatcctg tacgggccag atatccgcgt tgacattgat
     5041 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     5101 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     5161 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     5221 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     5281 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     5341 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     5401 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     5461 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     5521 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     5581 ggcgtgtacg gtgggaggtc tatataagca gagctctccc tatcagtgat agagatctcc
     5641 ctatcagtga tagagatcgt cgacgagctc gtttagtgaa ccgtcagatc gcctggagac
     5701 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggca
//
