LOCUS       sequence-478423-        7141 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Expresses C-FLAG tagged Rmp64 (CMV-RMP64-FLAG).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7141)
  AUTHORS   Che R, Mirani B, Panah M, Chen X, Luo H, Alexandrov A
  TITLE     Identification of Two Elusive Human Ribonuclease MRP-Specific 
            Protein Components.
  JOURNAL   bioRxiv [Preprint]. 2025 Jan 22:2025.01.19.633795. doi: 
            10.1101/2025.01.19.633795.
  PUBMED    39896489
REFERENCE   2  (bases 1 to 7141)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Jan 22:2025.01.19.633795. doi: 
            10.1101/2025.01.19.633795."
COMMENT     Sequence Label: CMV-RMP64-FLAG (pAVA3889)
FEATURES             Location/Qualifiers
     source          1..7141
                     /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             2629..2652
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2694..2712)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2695..2712)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2732..2749)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2738..2962
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3008..3436
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3095..3114)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3305..3326
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3445..3465)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3450..3779
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3630..3765
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3692..3711
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3846..4640
                     /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(3900..3919)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4510..4529
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4814..4935
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4851..4870)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4905..4924
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4984..5000)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4984..5000)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4997..5019)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5008..5024
                     /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(5032..5038,5039..5056,5057..5062))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5077..5098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5215..5232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5386..5974)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5466..5485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6145..6936,6937..7005))
                     /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     6768..6787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7006..7110)
                     /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 ccaccatgat ggctgcggtg ccgccgggcc tggagccgtg gaaccgtgtg
      961 agaatcccta aggcggggaa ccgcagcgca gtgacagtgc agaaccccgg cgcggccctt
     1021 gacctttgca ttgcagctgt aattaaagaa tgccatctcg tcatactgtc gctgaagagc
     1081 caaaccttag atgcagaaac agatgtgtta tgtgcagtcc tttacagcaa tcacaacaga
     1141 atgggccgcc acaaacccca tttggccctc aaacaggttg agcaatgttt aaagcgtttg
     1201 aaaaacatga atttggaggg ctcaattcaa gacctgtttg agttgttttc ttccaatgaa
     1261 aatcagccct taactaccaa agtatgtgtt gtccccagtc agccagtggt ggagttggtg
     1321 ttgatgaagg ttttgggagc ctgcaagttg ttgctccgct tgttggactg ctgctgcaaa
     1381 acttttcttt tgactgtgaa acatctaggt ttgcaagagt tcattatttt aaaccttgtg
     1441 atggttgggc tggtgagcag gttatgggtt ctctataaag gtgtcttaaa aaggttgatt
     1501 ttgttatatg agcctttgtt tggattgctt caagaggtcg ctaggattca accaatgcct
     1561 tacttcaaag attttacctt tccttctgat atcactgaat ttttaggaca gccatatttt
     1621 gaagccttta agaaaaaaat gcctatagct tttgcagcta aaggaataaa taaattgcta
     1681 aataaactgt ttttaataaa tgagcagtca ccaagagcca gtgaagaaac cttgcttgga
     1741 atttcaaaaa aagctaaaca aatgaagatc aatgtacaga ataatgtgga tcttggacag
     1801 ccagtaaaga ataagagagt cttcaaagaa gagtcatcag aatttgatgt gagggctttc
     1861 tgcaaccagc tgaaacacaa agctactcag gagaccagtt ttgattttaa atgttctcaa
     1921 tccagactaa agacaaccaa gtattcttct cagaaagtga taggaactcc tcatgccaaa
     1981 agttttgtgc aaagattccg agaggctgag tccttcacac aactttctga agaaatccag
     2041 atggcagttg tatggtgcag gagcaaaaaa ctcaaggctc aggccatttt tctgggtaac
     2101 aaacttctta aaagcaaccg gcttaaacat ctggaagctc aaggtactag tttgccaaag
     2161 aaactagagt gcataaaaac gtctatttgc aaccaccttc ttcgtggctc aggtatcaaa
     2221 acttcaaagc atcatctgag acagagaaga tcacagaata aatttttacg gagacaaagg
     2281 aaaccacaga gaaagttgca gtcgactctt ttaagggaaa ttcagcagtt ctctcaaggg
     2341 actcggaaga gtgctacaga taccagtgct aagtggagac tctcacactg tactgtgcat
     2401 agaactgatc tctaccctaa cagtaagcag ctcttgaata gtggagtttc aatgcctgtc
     2461 atacaaacta aggagaaaat gattcatgaa aatcttagag gcatccatga aaatgaaact
     2521 gattcgtgga cggtgatgca aataaataaa aacagtacat caggaaccat taaggagaca
     2581 gatgacattg atgatatttt tgctttaatg ggagttatcg atgtcgagga ttacaaggat
     2641 gacgacgata agtagggccc ggtaccctcg agcatgcatc tagagggccc tattctatag
     2701 tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     2761 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2821 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2881 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2941 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3001 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3061 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3121 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3181 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3241 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3301 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3361 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3421 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3481 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3541 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3601 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3661 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3721 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3781 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     3841 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3901 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3961 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4021 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4081 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4141 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4201 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4261 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4321 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4381 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4441 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4501 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4561 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4621 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4681 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4741 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4801 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4861 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4921 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4981 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5041 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5101 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5161 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5221 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5281 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5341 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5401 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5461 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5521 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5581 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5641 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5701 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5761 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5821 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5881 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5941 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6001 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6061 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6121 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6181 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6241 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6301 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6361 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6421 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6481 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6541 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6601 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6661 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6721 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6781 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6841 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6901 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6961 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7021 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7081 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7141 c
//
