LOCUS       sequence-239425-        6842 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of PNRC1 N-HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6842)
  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 6842)
  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 HA-PNRC1
FEATURES             Location/Qualifiers
     source          1..6842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        69..1247
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          300..1238
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1195..1215
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1264..1283
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1264..1282
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1314..1338
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1362..1388
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(2386..2410)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2436..2477
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2487..2504
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2527..2544)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2533..2757
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2803..3231
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2890..2909)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3100..3121
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3240..3260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3245..3574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3425..3560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3487..3506
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3622..3669
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3688..4086
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4244..4365
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4281..4300)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4335..4354
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4414..4430)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4414..4430)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4427..4449)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4438..4454
                     /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(4462..4468,4469..4486,4487..4492))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4507..4528
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4645..4662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4805..5393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4885..4904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5564..6355,6356..6424))
                     /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     6187..6206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6425..6529)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6604..6623)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag
       61 tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
      121 agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact
      181 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
      241 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
      301 taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc
      361 cttgaattac ttccacctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa
      421 gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg
      481 aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc
      541 tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt
      601 ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt
      661 ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg
      721 gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc
      781 tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt
      841 cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa
      901 aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg
      961 cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc
     1021 acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt
     1081 atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca gcttggcact
     1141 tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc
     1201 ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgagga attagcttgg
     1261 tactaatacg actcactata gggagaccca agctggctag gtaagcttga tcaacaagtt
     1321 tgtacaaaaa agcaggctcc gcggccgccc ccttcaccat gtacccatac gacgtcccag
     1381 actacgctac tgtcgtctcc gtcccgcagc gggagccgct cgtcctgggt ggccgccttg
     1441 cgccgcttgg cttttcctcc cgaggttact ttggggccct cccgatggtg accacggctc
     1501 cgcctccttt accccggatc ccggaccccc gggcactgcc cccgaccctc ttccttcctc
     1561 atttcctagg gggagatggc ccgtgtctga ccccccagcc tcgcgctcca gcagctctgc
     1621 ccaaccgcag cctcgccgtg gcgggaggca ctcctcgggc agcgccgaag aagcggcgaa
     1681 agaagaaggt gcgggccagc cccgcagggc agctgcccag ccgcttccac cagtaccagc
     1741 agcaccggcc gagtctggag ggcggccgga gccccgcgac cggcccgagc ggagcgcagg
     1801 aggtcccggg cccggccgcc gccttggccc cgagtcctgc agccgcagcc ggcacggagg
     1861 gagccagccc cgaccttgcc ccgctgcggc ccgcggctcc cggccaaacc cccctcagga
     1921 aagaggtttt aaaatcaaag atgggaaaat cggagaaaat tgcccttccc catggccagc
     1981 ttgttcatgg tatacacttg tatgagcaac caaagataaa cagacagaaa agcaaatata
     2041 acttgccact aaccaagatc acctctgcaa aaagaaatga aaacaacttt tggcaggatt
     2101 ctgtttcatc tgacagaatt cagaagcagg aaaaaaagcc ttttaaaaat accgagaaca
     2161 ttaaaaattc gcatttgaag aaatcagcat ttctaactga agtgagccaa aaggaaaatt
     2221 atgctggggc aaagtttagt gatccacctt ctcctagtgt tcttccaaag cctcctagtc
     2281 actggatggg aagcactgtt gaaaattcca accaaaacag ggagctgatg gcagtacact
     2341 taaaaacgct cctcaaagtt caaacttaga agggtgggcg cgccgaccca gctttcttgt
     2401 acaaagtggt tgatctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     2461 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
     2521 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2581 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2641 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2701 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2761 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2821 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2881 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2941 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3001 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3061 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3121 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3181 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3241 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3301 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3421 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3481 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3541 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3601 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     3661 taatacgaca aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga agaatccacc
     3721 ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc
     3781 gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt
     3841 actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc
     3901 aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga
     3961 cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt
     4021 gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag
     4081 ggctaagcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac
     4141 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4201 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4261 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4321 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4381 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4441 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4501 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4561 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4621 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4681 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4741 taacgcagga aagaacatgt gagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4801 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4861 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4921 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4981 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5041 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5101 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5161 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5221 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5281 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5341 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5401 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5461 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5521 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5581 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5641 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5701 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5761 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5821 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5881 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5941 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6001 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6061 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6121 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6181 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6241 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6301 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6361 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6421 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6481 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6541 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     6601 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     6661 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     6721 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     6781 cagatatacg cgttgacatt gattattgac taggcttttg caaaaagctt tgcaaagatg
     6841 ga
//
