LOCUS       sequence-311762-        6268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impacts 4 uracils in a poly-uracil tract in 
            conjunctiuon with an immediately upstream hairpin have on human 
            polymerase III transcription elongation from a U6 promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6268)
  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 6268)
  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-1_PolyU(4)
            _HairpinDistance(0...
FEATURES             Location/Qualifiers
     source          1..6268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..330)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      15..150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     315..335
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        363..611
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     363..383
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     534..553
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    1056..1177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   1226..1355
                     /label=AAV2 ITR
     rep_origin      complement(1449..1904)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1586..1607)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1798..1817
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1931..2035
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2036..2104,2105..2896)
                     /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(2254..2273)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3067..3655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3556..3575
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3924..4058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4483..4503)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4633..4698
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(5118..5912)
                     /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(5229..5248)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5839..5858
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
       61 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
      121 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
      181 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
      241 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
      301 tggggacttt ccacacccta actgacacac attccacagc cggatccaag gtcgggcagg
      361 aagagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
      421 agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
      481 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
      541 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
      601 gacgaaacac cgtgctcgct tcggcagcac atatactagt tggcggttgc ggctttctgc
      661 tgcaatcttt tcgacggcca tcagtcgccg gtcccaagcc cggataaaat gggagggggc
      721 gggaaaccgc ctaaccatgc cgactgatgg caggcccgga tagctcagtc ggtagagcag
      781 cggccgcgcg attcgaggaa ggaggtctga ggaggtcact gaatcgcgcg gccgcgggtc
      841 cagggttcaa gtccctgttc gggcctgcca tcagtcggcg tggactgtag aacactgcca
      901 atgccggtcc caagcccgga taaaagtgga gggtacagtc cacgctctag agcggacttc
      961 ggtccgcttt ttactaggac ctgcaggcat gcaagcttga cgtcggttac cgatatccat
     1021 atggcggccg catcgatctc gagccgcgga ctagtaactt gtttattgca gcttataatg
     1081 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1141 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtcttacgt agataagtag
     1201 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     1261 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     1321 gcccgggcgg cctcagtgag cgagcgagcg cgcagagagg gacagatctg cagatctgct
     1381 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     1441 gcgaatggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     1501 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     1561 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     1621 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     1681 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     1741 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     1801 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     1861 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     1921 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1981 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2041 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2101 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2161 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2221 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2281 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2341 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2401 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2461 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2521 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2581 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2641 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2701 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2761 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2821 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2881 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2941 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3001 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3061 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3121 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3181 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3241 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3301 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3361 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3421 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3481 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3541 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3601 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3661 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3721 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3781 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgagcgcg cagagaggga
     3841 gtggccaact ccatcactag gggttcctgg aggggtggag tcgtgacgtg aattacgtca
     3901 tagggttagg gaggtcctgg atcgatccag acatgataag atacattgat gagtttggac
     3961 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     4021 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     4081 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     4141 aatgtggtat ggctgattat gatctctagt caaggcacta tacatcaaat attccttatt
     4201 aaccccttta caaattaaaa agctaaaggt acacaatttt tgagcatagt tattaatagc
     4261 agacactcta tgcctgtgtg gagtaagaaa aaacagtatg ttatgattat aactgttatg
     4321 cctacttata aaggttacag aatatttttc cataattttc ttgtatagca gtgcagcttt
     4381 ttcctttgtg gtgtaaatag caaagcaagc aagagttcta ttactaaaca cagcatgact
     4441 caaaaaactt agcaattctg aaggaaagtc cttggggtct tctacctttc tcttcttttt
     4501 tggaggagta gaatgttgag agtcagcagt agcctcatca tcactagatg gcatttcttc
     4561 tgagcaaaac aggttttcct cattaaaggc attccaccac tgctcccatt catcagttcc
     4621 ataggttgga atctaaaata cacaaacaat tagaatcagt agtttaacac attatacact
     4681 taaaaatttt atatttacct tagagcttta aatctctgta ggtagtttgt ccaattatgt
     4741 cacaccacag aagtaaggtt ccttcacaaa gatccggacc aaagcggcca tcgtgcctcc
     4801 ccactcctgc agttcggggg catggatgcg cggatagccg ctgctggttt cctggatgcc
     4861 gacggatttg cactgccggt agaactccgc gaggtcgtcc agcctcaggc agcagctgaa
     4921 ccaactcgcg aggggatcga gcccggggtg ggcgaagaac tccagcatga gatccccgcg
     4981 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     5041 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     5101 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     5161 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     5221 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     5281 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     5341 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg
     5401 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     5461 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     5521 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     5581 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     5641 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
     5701 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg
     5761 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
     5821 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     5881 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
     5941 tcagatcttg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact
     6001 ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct
     6061 gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt
     6121 ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt
     6181 cagcaccgtt tctgcggact ggctttctac gtgttccgct tcctttagca gcccttgcgc
     6241 cctgagtgct tgcggcagcg tgaagctt
//
