LOCUS       sequence-478425-        6100 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expresses C-FLAG tagged Rmp24 (CMV-RMP24-FLAG).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6100)
  AUTHORS   Che R, Mirani B, Panah M, Chen X, Luo H, Alexandrov A
  TITLE     Identification of RMP24 and RMP64, human ribonuclease MRP-specific 
            protein components.
  JOURNAL   Cell Rep. 2025 Jun 24;44(6):115752. doi: 
            10.1016/j.celrep.2025.115752. Epub 2025 May 24.
  PUBMED    40413743
REFERENCE   2  (bases 1 to 6100)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2025-06-24"; volume: "44(6):115752. doi"; pages: " 
            10.1016/j.celrep.2025.115752. Epub 2025 May 24"
COMMENT     Sequence Label: CMV-RMP24-FLAG (pAVA3890)
FEATURES             Location/Qualifiers
     source          1..6100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1588..1611
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1653..1671)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1654..1671)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1691..1708)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1697..1921
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1967..2395
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2054..2073)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2264..2285
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2404..2424)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2409..2738
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2589..2724
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2651..2670
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2805..3599
                     /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(2859..2878)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3469..3488
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3773..3894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3810..3829)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3864..3883
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3943..3959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3943..3959)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3956..3978)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3967..3983
                     /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(3991..3997,3998..4015,4016..4021))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4036..4057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4174..4191)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4345..4933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4425..4444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5104..5895,5896..5964))
                     /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     5727..5746
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5965..6069)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccaccatgag acagaagcac taccttgagg ctgcagcgcg gggactgcac
      961 gacagctgcc cgggccaagc ccgctacctc ctctgggcct acacttcgtc gcacgatgat
     1021 aagagcactt ttgaagaaac gtgtccatac tgtttccagc tgttggttct ggataactct
     1081 cgagtgcgtc tcaaacccaa agccaggttg acacccaaaa tacagaaact tcttaatcga
     1141 gaagcgagaa actatacact cagttttaaa gaagcaaaaa tggtgaaaaa gttcaaagac
     1201 tccaaaagtg tattgttgat cacttgtaaa acatgcaaca gaacagtgaa acatcatggt
     1261 aaaagtagaa gctttgtgtc aacattgaag agcaatcctg ccactcctac aagtaaactc
     1321 agcctgaaga caccagagag aaggactgca aacccaaatc atgacatgtc tggctcgaaa
     1381 ggcaagagcc cagcatcggt tttcagaaca cctacatctg gacagtcagt atctacttgc
     1441 tcctcaaaga acaccagcaa aacaaagaaa cacttctctc aactaaaaat gttacttagt
     1501 cagaatgaat cccaaaagat tccaaaggtg gacttcagaa atttcttatc ttctctgaag
     1561 ggtggacttt taaaaatcga tgtcgaggat tacaaggatg acgacgataa gtagggcccg
     1621 gtaccctcga gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct
     1681 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1741 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1801 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1861 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     1921 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     1981 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2041 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2101 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2161 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2221 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2281 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2341 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2401 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2461 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2521 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2581 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2641 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     2701 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     2761 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     2821 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     2881 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     2941 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3001 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3061 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3121 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3181 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3241 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3301 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3361 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3421 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3481 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3541 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3601 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     3661 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     3721 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     3781 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3841 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3901 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     3961 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4021 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4081 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4141 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4201 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4261 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4321 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4381 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4441 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4501 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4561 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4621 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4681 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4741 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4801 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4861 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4921 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4981 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5041 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5101 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5161 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5221 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5281 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5341 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5401 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5461 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5521 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5581 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5641 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5701 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5761 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5821 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5881 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5941 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6001 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6061 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
