LOCUS       sequence-250991-        8415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 5S-F30-Mango IV rRNA from a mU6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8415)
  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 8415)
  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-Mango IV
FEATURES             Location/Qualifiers
     source          1..8415
                     /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        3281..3584
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3585..3788
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3822..4418
                     /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(3822..3841)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4318..4338
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4428..4481
                     /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      4488..4497
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4494..5204
                     /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(4639..4657)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4907..4926
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(5224..5257)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5313..5901
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5366..5386)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5784..5795)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5904..5920)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5905..5921)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6430..6610
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6672..7260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6752..6771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7431..8222,8223..8291))
                     /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     8054..8073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8292..8396)
                     /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 gtgggtaccg agggagtggt gaggatgagg cgagtaccca
     3121 catactctga tgatccttcg ggatcattca tggcaatttt tttctcgagt actaggatcc
     3181 attaggcggc cgcgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     3241 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     3301 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     3361 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     3421 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     3481 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     3541 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     3601 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     3661 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     3721 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     3781 gcagagctgg tttagtgaac cgtcagatcc gctagcccac catgaccgag tacaagccca
     3841 cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt
     3901 tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca
     3961 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     4021 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     4081 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     4141 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     4201 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     4261 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     4321 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     4381 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccgg atcgggagag ggcagaggaa
     4441 gtctgctaac atgcggtgac gtcgaggaga atcctggccc accggtcgcc accatggtga
     4501 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     4561 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     4621 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     4681 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     4741 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     4801 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     4861 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     4921 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     4981 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     5041 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     5101 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     5161 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagaattc gtcgagggac
     5221 ctaataactt cgtatagcat acattatacg aagttataca tgtttaaggg ttccggttcc
     5281 actaggtaca attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     5341 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     5401 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     5461 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     5521 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     5581 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     5641 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     5701 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     5761 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     5821 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     5881 ccctttgggc cgcctccccg catcgatacc gtcgacctcg atcgagacct agaaaaacat
     5941 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     6001 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     6061 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     6121 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     6181 ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga
     6241 tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag
     6301 aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta
     6361 ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg
     6421 gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6481 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6541 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6601 atctctagca gcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6661 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6721 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6781 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6841 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6901 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6961 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7021 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7081 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     7141 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7201 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7261 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7321 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7381 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7441 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7501 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7561 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7621 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7681 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7741 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7801 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7861 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7921 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7981 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8041 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8101 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8161 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8221 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8281 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8341 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8401 tttccccgaa aagtg
//
