LOCUS       sequence-250987-        8388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 5S-F30-Control rRNA from a mU6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8388)
  AUTHORS   Autour A, C Y Jeng S, D Cawte A, Abdolahzadeh A, Galli A, 
            Panchapakesan SSS, Rueda D, Ryckelynck M, Unrau PJ
  TITLE     Fluorogenic RNA Mango aptamers for imaging small non-coding RNAs in 
            mammalian cells.
  JOURNAL   Nat Commun. 2018 Feb 13;9(1):656. doi: 10.1038/s41467-018-02993-8.
  PUBMED    29440634
REFERENCE   2  (bases 1 to 8388)
  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: "Nat 
            Commun."; date: "2018-02-13"; volume: "9(1):656. doi"; pages: " 
            10.1038/s41467-018-02993-8"
COMMENT     Sequence Label: pSLQ-5S-F30-Control
FEATURES             Location/Qualifiers
     source          1..8388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        247..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        628..826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             844..1024
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1071..1196
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1693..1926
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2111..2155
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2453..2570
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2823..2856)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2860..2880
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     enhancer        3254..3557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3558..3761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3795..4391
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3795..3814)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4291..4311
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4401..4454
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      4461..4470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4467..5177
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4612..4630)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4880..4899
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(5197..5230)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5286..5874
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5339..5359)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5757..5768)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5877..5893)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5878..5894)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6403..6583
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6645..7233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6725..6744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7404..8195,8196..8264))
                     /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     8027..8046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8265..8369)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      841 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      901 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      961 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     1021 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     1081 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     1141 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     1201 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1261 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1321 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1381 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1441 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1501 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg
     1561 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     1621 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     1681 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     1741 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     1801 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     1861 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     1921 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     1981 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     2041 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     2101 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     2161 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     2221 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     2281 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     2341 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     2401 atcggcactg cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa
     2461 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     2521 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     2581 acagggacag cagagatcca gtttggttag taccgggccc gctctagaga tccgacgcgc
     2641 catctctagg cccgcgccgg ccccctcgca cggacttgtg ggagaagctc ggctactccc
     2701 ctgccccggt taatttgcat ataatatttc ctagtaacta tagaggctta atgtgcgata
     2761 aaagacagat aatctgttct ttttaatact agctacattt tacatgatag gcttggattt
     2821 ctataacttc gtatagcata cattatacga agttataaac agcacaaaag gaaactcacc
     2881 ctaactgtaa agtaattgtg tgttttgaga ctataagtat cccttggaga accaccttgt
     2941 tgggtctacg gccataccac cctgaacgcg cccgatctcg tctgatctcg gaagctaagc
     3001 agggtcgggc ctggttagta cttggatggg agaccgcctg ggaataccgg gtgctgtagg
     3061 cgtcgacttg ccatgtgtat gtggggaaac ccacatactc tgatgatcct tcgggatcat
     3121 tcatggcaat ttttttctcg agtactagga tccattaggc ggccgcgtgg ataaccgtat
     3181 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     3241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     3301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     3361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     3421 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     3481 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     3541 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     3601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     3661 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     3721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     3781 tccgctagcc caccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     3841 tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     3901 ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     3961 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     4021 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     4081 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     4141 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     4201 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     4261 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     4321 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     4381 agcccggtgc cggatcggga gagggcagag gaagtctgct aacatgcggt gacgtcgagg
     4441 agaatcctgg cccaccggtc gccaccatgg tgagcaaggg cgaggaggat aacatggcca
     4501 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     4561 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     4621 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     4681 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     4741 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     4801 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     4861 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     4921 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     4981 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     5041 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     5101 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     5161 acgagctgta caagtaagaa ttcgtcgagg gacctaataa cttcgtatag catacattat
     5221 acgaagttat acatgtttaa gggttccggt tccactaggt acaattcgat atcaagctta
     5281 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5341 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5401 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5461 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5521 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5581 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     5641 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     5701 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5761 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5821 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     5881 accgtcgacc tcgatcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     5941 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     6001 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     6061 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc
     6121 cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca
     6181 gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     6241 caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc
     6301 ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta
     6361 gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac
     6421 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     6481 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6541 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagcatgtg agcaaaaggc
     6601 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6661 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6721 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6781 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6841 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6901 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6961 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7021 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7081 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7141 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7201 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7261 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7321 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7381 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7441 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7501 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7561 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7621 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7681 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7741 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7801 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7861 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7921 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7981 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8041 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8101 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8161 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8221 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8281 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8341 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtg
//
