LOCUS       sequence-311746-        6246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 1 uracil in a poly-uracil tract on human 
            polymerase III transcription from a U6 promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6246)
  AUTHORS   Verosloff MS, Corcoran WK, Dolberg TB, Bushhouse DZ, Leonard JN, 
            Lucks JB
  TITLE     RNA Sequence and Structure Determinants of Pol III Transcriptional 
            Termination in Human Cells.
  JOURNAL   J Mol Biol. 2021 Jun 25;433(13):166978. doi: 
            10.1016/j.jmb.2021.166978. Epub 2021 Apr 1.
  PUBMED    33811918
REFERENCE   2  (bases 1 to 6246)
  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: "J Mol 
            Biol."; date: "2021-06-25"; volume: "433(13):166978. doi"; pages: " 
            10.1016/j.jmb.2021.166978. Epub 2021 Apr 1"
COMMENT     Sequence Label:  pAV-U6+27-Linear-2_PolyU(1)_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     307..327
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        355..603
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     355..375
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     526..545
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    1026..1147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   1196..1325
                     /label=AAV2 ITR
     rep_origin      complement(1419..1874)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1556..1577)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1768..1787
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1901..2005
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2006..2074,2075..2866)
                     /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(2224..2243)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3037..3625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3526..3545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3894..4028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4453..4473)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4603..4668
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(5088..5882)
                     /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(5199..5218)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5809..5828
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6239..6246,1..322))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
       61 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
      121 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
      181 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
      241 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
      301 ttccacaccc taactgacac acattccaca gccggatcca aggtcgggca ggaagagggc
      361 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt
      421 agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa
      481 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt
      541 accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac
      601 accgtgctcg cttcggcagc acatatacta gtgattgatg tcgacggcca tcagtcgccg
      661 gtcccaagcc cggataaaat gggagggggc gggaaaccgc ctaaccatgc cgactgatgg
      721 caggcccgga tagctcagtc ggtagagcag cggccgcgcg attcgaggaa ggaggtctga
      781 ggaggtcact gaatcgcgcg gccgcgggtc cagggttcaa gtccctgttc gggcctgcca
      841 tcagtcggcg tggactgtag aacactgcca atgccggtcc caagcccgga taaaagtgga
      901 gggtacagtc cacgctctag agcggacttc ggtccgcttt ttactaggac ctgcaggcat
      961 gcaagcttga cgtcggttac cgatatccat atggcggccg catcgatctc gagccgcgga
     1021 ctagtaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1081 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1141 tatcttatca tgtcttacgt agataagtag catggcgggt taatcattaa ctacaaggaa
     1201 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     1261 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     1321 cgcagagagg gacagatctg cagatctgct ggcgtaatag cgaagaggcc cgcaccgatc
     1381 gcccttccca acagttgcgc agcctgaatg gcgaatggaa attgtaagcg ttaatatttt
     1441 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     1501 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     1561 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     1621 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     1681 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     1741 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     1801 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     1861 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1921 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1981 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2041 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2101 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2161 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2221 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2281 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2341 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2401 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2461 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2521 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2581 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2641 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2701 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2761 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2821 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2881 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2941 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3001 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3061 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3121 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3181 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3241 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3301 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3361 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3421 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3481 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3541 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3601 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3661 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3721 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3781 cgagtcagtg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgg
     3841 aggggtggag tcgtgacgtg aattacgtca tagggttagg gaggtcctgg atcgatccag
     3901 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3961 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     4021 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     4081 aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat gatctctagt
     4141 caaggcacta tacatcaaat attccttatt aaccccttta caaattaaaa agctaaaggt
     4201 acacaatttt tgagcatagt tattaatagc agacactcta tgcctgtgtg gagtaagaaa
     4261 aaacagtatg ttatgattat aactgttatg cctacttata aaggttacag aatatttttc
     4321 cataattttc ttgtatagca gtgcagcttt ttcctttgtg gtgtaaatag caaagcaagc
     4381 aagagttcta ttactaaaca cagcatgact caaaaaactt agcaattctg aaggaaagtc
     4441 cttggggtct tctacctttc tcttcttttt tggaggagta gaatgttgag agtcagcagt
     4501 agcctcatca tcactagatg gcatttcttc tgagcaaaac aggttttcct cattaaaggc
     4561 attccaccac tgctcccatt catcagttcc ataggttgga atctaaaata cacaaacaat
     4621 tagaatcagt agtttaacac attatacact taaaaatttt atatttacct tagagcttta
     4681 aatctctgta ggtagtttgt ccaattatgt cacaccacag aagtaaggtt ccttcacaaa
     4741 gatccggacc aaagcggcca tcgtgcctcc ccactcctgc agttcggggg catggatgcg
     4801 cggatagccg ctgctggttt cctggatgcc gacggatttg cactgccggt agaactccgc
     4861 gaggtcgtcc agcctcaggc agcagctgaa ccaactcgcg aggggatcga gcccggggtg
     4921 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     4981 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     5041 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     5101 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     5161 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     5221 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     5281 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     5341 ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     5401 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     5461 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     5521 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     5581 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     5641 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc
     5701 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     5761 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     5821 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     5881 atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc
     5941 cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc
     6001 gccccagctg gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc
     6061 catgtaagcc cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag
     6121 atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac
     6181 gtgttccgct tcctttagca gcccttgcgc cctgagtgct tgcggcagcg tgaagctttt
     6241 tgcaaa
//
