LOCUS       sequence-250989-        8390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mgU2-47 scaRNA from a mU6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8390)
  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 8390)
  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-mgU2-47
FEATURES             Location/Qualifiers
     source          1..8390
                     /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        3256..3559
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3560..3763
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3797..4393
                     /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(3797..3816)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4293..4313
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4403..4456
                     /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      4463..4472
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4469..5179
                     /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(4614..4632)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4882..4901
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(5199..5232)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5288..5876
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5341..5361)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5759..5770)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5879..5895)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5880..5896)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6405..6585
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6647..7235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6727..6746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7406..8197,8198..8266))
                     /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     8029..8048
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8267..8371)
                     /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 tggagcaaag tgatgagtaa tactggctgg agccccaaag aggcacgtgt gtgtgtttgt
     3001 gtgtgtgtgt atatgcttgt cagtgcatgc acgtgtatgt ctgggagtac aaatgggtgc
     3061 gactggttgt agggaactag ctatgtgcct tctattaggc catgacagtc aaactgataa
     3121 gatctgattg ctttttttct cgagtactag gatccattag gcggccgcgt ggataaccgt
     3181 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
     3241 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     3301 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     3361 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     3421 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     3481 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     3541 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     3601 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     3661 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     3721 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     3781 gatccgctag cccaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     3841 cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     3901 caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     3961 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     4021 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     4081 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     4141 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     4201 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     4261 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     4321 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     4381 caagcccggt gccggatcgg gagagggcag aggaagtctg ctaacatgcg gtgacgtcga
     4441 ggagaatcct ggcccaccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc
     4501 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     4561 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     4621 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     4681 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     4741 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     4801 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     4861 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     4921 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     4981 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     5041 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     5101 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     5161 ggacgagctg tacaagtaag aattcgtcga gggacctaat aacttcgtat agcatacatt
     5221 atacgaagtt atacatgttt aagggttccg gttccactag gtacaattcg atatcaagct
     5281 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     5341 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     5401 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     5461 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5521 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5581 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5641 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     5701 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5761 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5821 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     5881 ataccgtcga cctcgatcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     5941 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     6001 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     6061 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata
     6121 tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag aactacacac
     6181 cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
     6241 agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga
     6301 gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc
     6361 tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag
     6421 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     6481 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     6541 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag
     6601 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6661 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6721 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6781 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6841 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6901 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6961 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7021 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7081 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7141 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7201 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7261 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7321 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7381 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7441 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7501 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7561 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7621 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7681 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7741 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7801 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7861 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7921 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7981 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8041 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8101 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8161 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8221 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8281 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     8341 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
//
