LOCUS       sequence-239378-        7540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of PNRC1 N-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7540)
  AUTHORS   Gaviraghi M, Vivori C, Pareja Sanchez Y, Invernizzi F, Cattaneo A, 
            Santoliquido BM, Frenquelli M, Segalla S, Bachi A, Doglioni C, 
            Pelechano V, Cittaro D, Tonon G
  TITLE     Tumor suppressor PNRC1 blocks rRNA maturation by recruiting the 
            decapping complex to the nucleolus.
  JOURNAL   EMBO J. 2018 Dec 3;37(23). pii: embj.201899179. doi: 
            10.15252/embj.201899179. Epub 2018 Oct 29.
  PUBMED    30373810
REFERENCE   2  (bases 1 to 7540)
  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: "EMBO J."; 
            date: "2018-12-3"; volume: "37(23). pii: embj.201899179. doi"; 
            pages: " 10.15252/embj.201899179. Epub 2018 Oct 29"
COMMENT     Sequence Label: pEF-DEST51 GFP-PNRC1
FEATURES             Location/Qualifiers
     source          1..7540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        31..1209
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          262..1200
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1157..1177
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1226..1245
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1226..1244
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1276..1300
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1311..1313,1314..1316,1317..2027)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1311..1313,1314..1316,1317..2027)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1356..1377)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1617..1636)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1964..1985
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3046..3070)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3096..3137
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3147..3164
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3187..3204)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3193..3417
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3463..3891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3550..3569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3760..3781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3900..3920)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3905..4234
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4085..4220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4147..4166
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4282..4329
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4348..4746
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4904..5025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4941..4960)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4995..5014
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5074..5090)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5074..5090)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5087..5109)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5098..5114
                     /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(5122..5128,5129..5146,5147..5152))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5167..5188
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5305..5322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5465..6053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5545..5564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6224..7015,7016..7084))
                     /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     6847..6866
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7085..7189)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7264..7283)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc
       61 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag
      121 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
      181 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
      241 gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt
      301 acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta cgtgattctt
      361 gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc
      421 ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc
      481 tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt
      541 tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat
      601 ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag
      661 cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag
      721 tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc
      781 tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc
      841 ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag
      901 tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg
      961 gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct
     1021 ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact
     1081 gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt
     1141 ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag
     1201 gtgtcgtgag gaattagctt ggtactaata cgactcacta tagggagacc caagctggct
     1261 aggtaagctt gatcaacaag tttgtacaaa aaagcaggct ccgcggccgc atggtgagca
     1321 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1381 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1441 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1501 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     1561 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1621 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1681 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1741 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1801 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1861 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1921 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1981 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggcg gccgccccct
     2041 tcaccatgac tgtcgtctcc gtcccgcagc gggagccgct cgtcctgggt ggccgccttg
     2101 cgccgcttgg cttttcctcc cgaggttact ttggggccct cccgatggtg accacggctc
     2161 cgcctccttt accccggatc ccggaccccc gggcactgcc cccgaccctc ttcctccctc
     2221 atttcctagg gggagatggc ccgtgtctga ccccccagcc tcgcgctcca gcagctctgc
     2281 ccaaccgcag cctcgccgtg gcgggaggca ctcctcgggc agcgccgaag aagcggcgaa
     2341 agaagaaggt gcgggccagc cccgcagggc agctgcccag ccgcttccac cagtaccagc
     2401 agcaccggcc gagtctggag ggcggccgga gccccgcgac cggcccgagc ggagcgcagg
     2461 aggtcccggg cccggccgcc gccttggccc cgagtcctgc agccgcagcc ggcacggagg
     2521 gagccagccc cgaccttgcc ccgctgcggc ccgcggctcc cggccaaacc cccctcagga
     2581 aagaggtttt aaaatcaaag atgggaaaat cggagaaaat tgcccttccc catggccagc
     2641 ttgttcatgg tatacacttg tatgagcaac caaagataaa cagacagaaa agcaaatata
     2701 acttgccact aaccaagatc acctctgcaa aaagaaatga aaacaacttt tggcaggatt
     2761 ctgtttcatc tgacagaatt cagaagcagg aaaaaaagcc ttttaaaaat accgagaaca
     2821 ttaaaaattc gcatttgaag aaatcagcat ttctaactga agtgagccaa aaggaaaatt
     2881 atgctggggc aaagtttagt gatccacctt ctcctagtgt tcttccaaag cctcctagtc
     2941 actggatggg aagcactgtt gaaaattcca accaaaacag ggagctgatg gcagtacact
     3001 taaaaacgct cctcaaagtt caaacttaga agggtgggcg cgccgaccca gctttcttgt
     3061 acaaagtggt tgatctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     3121 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
     3181 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3241 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3301 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3361 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3421 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3481 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3541 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3601 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3661 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3721 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3781 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3841 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3901 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4021 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4081 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4141 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4201 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4261 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     4321 taatacgaca aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga agaatccacc
     4381 ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc
     4441 gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt
     4501 actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc
     4561 aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga
     4621 cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt
     4681 gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag
     4741 ggctaagcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac
     4801 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4861 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4921 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4981 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5041 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5101 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5161 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5221 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5281 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5341 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5401 taacgcagga aagaacatgt gagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5461 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5521 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5581 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5641 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5701 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5761 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5821 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5881 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5941 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6001 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6061 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6121 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6181 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6241 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6301 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6361 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6421 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6481 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6541 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6601 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6661 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6721 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6781 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6841 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6901 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6961 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7021 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7081 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7141 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7201 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     7261 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7321 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     7381 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     7441 cagatatacg cgttgacatt gattattgac taggcttttg caaaaagctt tgcaaagatg
     7501 gataaagttt taaacagaga ggaatctttg cagctaatgg
//
