LOCUS       sequence-478424-        6037 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Expresses C-FLAG tagged Rpp25 (CMV-RPP25-FLAG).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6037)
  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 6037)
  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-RPP25-FLAG (pAVA3895)
FEATURES             Location/Qualifiers
     source          1..6037
                     /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             1525..1548
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1590..1608)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1591..1608)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1628..1645)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1634..1858
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1904..2332
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1991..2010)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2201..2222
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2341..2361)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2346..2675
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2526..2661
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2588..2607
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2742..3536
                     /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(2796..2815)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3406..3425
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3710..3831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3747..3766)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3801..3820
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3880..3896)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3880..3896)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3893..3915)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3904..3920
                     /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(3928..3934,3935..3952,3953..3958))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3973..3994
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4111..4128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4282..4870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4362..4381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5041..5832,5833..5901))
                     /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     5664..5683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5902..6006)
                     /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 ccaccatgga gaacttccgt aaggtgcgct ccgaagaggc gccagcgggg
      961 tgcggggccg agggaggcgg cccgggctcc ggccccttcg cagacctggc gccgggcgcg
     1021 gtgcacatgc gggtcaagga aggcagcaag atccggaacc tgatggcctt cgccaccgcc
     1081 agcatggcgc agccagccac gcgcgccatc gtcttcagcg gctgcggccg ggccaccacc
     1141 aaaaccgtca cgtgcgccga gatcctcaag cgccgcctgg cgggcctgca ccaggtcacg
     1201 cggctgcgct accggagcgt acgcgaggtg tggcagagcc tcccgcctgg gcccacgcag
     1261 ggtcagacgc ctggcgagcc ggccgctagt ctcagcgtac ttaagaacgt gcccggcctc
     1321 gccatcctac tttccaagga cgcgctggat ccgcgacagc ccggctacca gcccccgaat
     1381 ccccatcctg gtccctcgtc cccgccagcc gcgccagcgt ccaagaggag cctaggggaa
     1441 cccgcagctg gagaaggctc cgcgaagcga tcgcaacccg agccaggggt tgcggacgag
     1501 gatcagacgg ccatcgatgt cgaggattac aaggatgacg acgataagta gggcccggta
     1561 ccctcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc
     1621 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1681 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1741 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1801 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     1861 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     1921 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1981 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2041 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2101 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2161 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2221 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2281 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     2341 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2401 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2461 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2521 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2581 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     2641 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     2701 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     2761 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     2821 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     2881 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     2941 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3001 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3061 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3121 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3181 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3241 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     3301 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3361 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3421 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3481 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3541 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3601 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3661 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3721 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3781 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3841 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3901 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3961 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4021 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4081 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4141 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4201 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4261 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4321 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4381 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4441 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4501 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4561 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4621 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4681 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4741 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4801 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4861 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4921 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4981 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5041 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5101 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5161 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5221 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5281 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5341 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5401 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5461 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5521 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5581 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5641 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5701 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5761 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5821 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5881 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5941 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6001 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
