LOCUS       sequence-451662-        6072 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express tRNA-Corn as a reporter of U6 promoter activity in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6072)
  AUTHORS   Song W, Filonov GS, Kim H, Hirsch M, Li X, Moon JD, Jaffrey SR
  TITLE     Imaging RNA polymerase III transcription using a photostable 
            RNA-fluorophore complex.
  JOURNAL   Nat Chem Biol. 2017 Nov;13(11):1187-1194. doi: 
            10.1038/nchembio.2477. Epub 2017 Sep 25.
  PUBMED    28945233
REFERENCE   2  (bases 1 to 6072)
  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"; doi: 
            "10.1038/nchembio.2477"; journalName: "Nat Chem Biol"; date: 
            "2017-11"; volume: "13"; issue: "11"; pages: "1187-1194"
COMMENT     Sequence Label: pAV-U6+27-tCorn
FEATURES             Location/Qualifiers
     source          1..6072
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..20)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     611..630
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1041..1370)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1055..1190
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1355..1375
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1403..1651
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1403..1423
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1574..1593
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    1896..2017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2066..2195
                     /label=AAV2 ITR
     rep_origin      complement(2289..2744)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2426..2447)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2638..2657
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2771..2875
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2876..2944,2945..3736)
                     /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     complement(3094..3113)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3907..4495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4396..4415
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4768..4902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5327..5347)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5477..5542
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(join(5962..6072,1..684))
                     /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"
ORIGIN
        1 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
       61 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
      121 atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc
      181 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
      241 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
      301 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
      361 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
      421 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
      481 cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac
      541 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
      601 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
      661 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatct
      721 tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg
      781 cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa
      841 aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc
      901 gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg
      961 tttctgcgga ctggctttct acgtgttccg cttcctttag cagcccttgc gccctgagtg
     1021 cttgcggcag cgtgaagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     1081 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt
     1141 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
     1201 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
     1261 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
     1321 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacagc
     1381 cggatccaag gtcgggcagg aagagggcct atttcccatg attccttcat atttgcatat
     1441 acgatacaag gctgttagag agataattag aattaatttg actgtaaaca caaagatatt
     1501 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
     1561 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
     1621 tttatatatc ttgtggaaag gacgaaacac cgtgctcgct tcggcagcac atatactagt
     1681 cgacgcccgg atagctcagt cggtagagca gcggccgcga ggaaggaggt ctgaggaggt
     1741 cactgcggcc gcgggtccag ggttcaagtc cctgttcggg cgccatctag agcggacttc
     1801 ggtccgcttt ttactaggac ctgcaggcat gcaagcttga cgtcggttac cgatatccat
     1861 atggcggccg catcgatctc gagccgcgga ctagtaactt gtttattgca gcttataatg
     1921 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1981 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtcttacgt agataagtag
     2041 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     2101 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     2161 gcccgggcgg cctcagtgag cgagcgagcg cgcagagagg gacagatctg cagatctgct
     2221 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2281 gcgaatggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2341 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2401 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2461 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2521 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2581 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2641 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     2701 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     2761 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2821 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2881 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2941 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3001 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3061 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3121 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3181 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3241 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3301 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3361 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3421 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3481 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3541 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3601 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3661 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3721 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3781 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3841 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3901 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3961 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4021 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4081 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4141 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4201 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4261 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4321 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4381 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4441 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4501 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4561 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4621 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgagcgag cgcgcagaga
     4681 gggagtggcc aactccatca ctaggggttc ctggaggggt ggagtcgtga cgtgaattac
     4741 gtcatagggt tagggaggtc ctggatcgat ccagacatga taagatacat tgatgagttt
     4801 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     4861 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     4921 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     4981 tacaaatgtg gtatggctga ttatgatctc tagtcaaggc actatacatc aaatattcct
     5041 tattaacccc tttacaaatt aaaaagctaa aggtacacaa tttttgagca tagttattaa
     5101 tagcagacac tctatgcctg tgtggagtaa gaaaaaacag tatgttatga ttataactgt
     5161 tatgcctact tataaaggtt acagaatatt tttccataat tttcttgtat agcagtgcag
     5221 ctttttcctt tgtggtgtaa atagcaaagc aagcaagagt tctattacta aacacagcat
     5281 gactcaaaaa acttagcaat tctgaaggaa agtccttggg gtcttctacc tttctcttct
     5341 tttttggagg agtagaatgt tgagagtcag cagtagcctc atcatcacta gatggcattt
     5401 cttctgagca aaacaggttt tcctcattaa aggcattcca ccactgctcc cattcatcag
     5461 ttccataggt tggaatctaa aatacacaaa caattagaat cagtagttta acacattata
     5521 cacttaaaaa ttttatattt accttagagc tttaaatctc tgtaggtagt ttgtccaatt
     5581 atgtcacacc acagaagtaa ggttccttca caaagatccg gaccaaagcg gccatcgtgc
     5641 ctccccactc ctgcagttcg ggggcatgga tgcgcggata gccgctgctg gtttcctgga
     5701 tgccgacgga tttgcactgc cggtagaact ccgcgaggtc gtccagcctc aggcagcagc
     5761 tgaaccaact cgcgagggga tcgagcccgg ggtgggcgaa gaactccagc atgagatccc
     5821 cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat
     5881 agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca
     5941 tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
     6001 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
     6061 aagctcttca gc
//
