LOCUS       sequence-134202-        6694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression clone for human NHP2L1 (15.5kD) with C-terminal 
            3xFLAG-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6694)
  AUTHORS   Falaleeva M, Pages A, Matuszek Z, Hidmi S, Agranat-Tamir L, Korotkov
            K, Nevo Y, Eyras E, Sperling R, Stamm S
  TITLE     Dual function of C/D box small nucleolar RNAs in rRNA modification 
            and alternative pre-mRNA splicing.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Mar 22;113(12):E1625-34. doi: 
            10.1073/pnas.1519292113. Epub 2016 Mar 8.
  PUBMED    26957605
REFERENCE   2  (bases 1 to 6694)
  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: 
            "10.1073/pnas.1519292113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-03-22- 22"; volume: "113"; issue: "12"; pages: 
            "E1625-34"
COMMENT     Sequence Label: pCMV14_NHP2L1(15.5kD)
FEATURES             Location/Qualifiers
     source          1..6694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(735..1526,1527..1595))
                     /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     1358..1377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1596..1700)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1727..2182
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1814..1833)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2024..2045
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2308..2330
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2322..2339
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2323..2339
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        2500..2879
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2880..3083
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3034..3054
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3559..3624
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3635..4257
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3834..3853)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4281..4301)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4299..4596
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4650..5444
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4704..4723)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5314..5333
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6096..6230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6133..6152)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6187..6206
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6235..6254)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6236..6254)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6268..6284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6268..6284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6281..6303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6292..6308
                     /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(6316..6322,6323..6340,6341..6346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6361..6382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6499..6516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6670..6694,1..564))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
       61 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      121 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      181 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
      241 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      301 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      361 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
      421 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      481 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      541 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      601 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
      661 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
      721 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
      781 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
      841 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
      901 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
      961 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     1021 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     1081 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     1141 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     1201 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     1261 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     1321 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     1381 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     1441 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     1501 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     1561 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     1621 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     1681 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     1741 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1801 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1861 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1921 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1981 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2041 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2101 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2161 aaaatattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc
     2221 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     2281 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2341 ccaagctgat ctatacattg aatcaatatt ggcaattagc catattagtc attggttata
     2401 tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt
     2461 acatttatat tggctcatgt ccaatatgac cgccatgttg acattgatta ttgactagtt
     2521 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     2581 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     2641 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     2701 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc
     2761 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     2821 ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     2881 tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc
     2941 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     3001 ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt
     3061 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gaattaagct tgcggccgcg
     3121 aattcatcga tagatctgat atcggtacca gtcgacatga ctgaggctga tgtgaatcca
     3181 aaggcctatc cccttgccga tgcccacctc accaagaagc tactggacct cgttcagcag
     3241 tcatgtaact ataagcagct tcggaaagga gccaatgagg ccaccaaaac cctcaacagg
     3301 ggcatctctg agttcatcgt gatggctgca gacgccgagc cactggagat cattctgcac
     3361 ctgccgctgc tgtgtgaaga caagaatgtg ccctacgtgt ttgtgcgctc caagcaggcc
     3421 ctggggagag cctgtggggt ctccaggcct gtcatcgcct gttctgtcac catcaaagaa
     3481 ggctcgcagc tgaaacagca gatccaatcc attcagcagt ccattgaaag gctcttagtc
     3541 tctagaggat cccgggctga ctacaaagac catgacggtg attataaaga tcatgacatc
     3601 gactacaagg atgacgatga caagtagtga tcccgggtgg catccctgtg acccctcccc
     3661 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     3721 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     3781 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     3841 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     3901 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     3961 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     4021 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     4081 ggcgtgaacc actgctccct tccctgtcct tctgatttta aaataactat accagcagga
     4141 ggacgtccag acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg
     4201 cttgcttggc actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg
     4261 tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4321 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4381 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4441 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4501 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4561 tgagctattc cagaagtagt gaggaggctt ttttggagga attgatcagc ttgggatctg
     4621 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4681 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4741 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4801 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4861 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4921 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4981 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5041 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5101 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5161 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5221 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5281 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5341 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5401 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5461 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5521 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5581 gcgcggggat ctcatgctgg agttcttcgc ccaccccggg ctcgatcccc tcgcgagttg
     5641 gttcagctgc tgcctgaggc tggacgacct cgcggagttc taccggcagt gcaaatccgt
     5701 cggcatccag gaaaccagca gcggctatcc gcgcatccat gcccccgaac tgcaggagtg
     5761 gggaggcacg atggccgctt tggtcgaccc ggacgggacg ctcctgcgcc tgatacagaa
     5821 cgaattgctt gcaggcatct catgagtgtg tcttcccgtt ttccgcctga ggtcactgcg
     5881 tggatggagc gctggcgcct gctgcgcgac ggcgagctgc tcaccaccca ctcgccaagc
     5941 tggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccg atcataatca
     6001 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     6061 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     6121 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6181 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aattccctat
     6241 agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     6301 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6361 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6421 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6481 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6541 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6601 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6661 gctggcgttt ttccataggc tccgcccccc tgac
//
