LOCUS       sequence-311769-        6245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 4 uracils 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 6245)
  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 6245)
  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)_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    671..792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   841..970
                     /label=AAV2 ITR
     rep_origin      complement(1064..1519)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1201..1222)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1413..1432
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1546..1650
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1651..1719,1720..2511)
                     /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(1869..1888)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2682..3270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3171..3190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3539..3673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4098..4118)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4248..4313
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4733..5527)
                     /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(4844..4863)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5454..5473
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5884..6213)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5898..6033
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6198..6218
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg tgctcgcttc ggcagcacat atactagttg gcttttcgac ggccatcagt
      301 cgccggtccc aagcccggat aaaatgggag ggggcgggaa accgcctaac catgccgact
      361 gatggcaggc ccggatagct cagtcggtag agcagcggcc gcgcgattcg aggaaggagg
      421 tctgaggagg tcactgaatc gcgcggccgc gggtccaggg ttcaagtccc tgttcgggcc
      481 tgccatcagt cggcgtggac tgtagaacac tgccaatgcc ggtcccaagc ccggataaaa
      541 gtggagggta cagtccacgc tctagagcgg acttcggtcc gctttttact aggacctgca
      601 ggcatgcaag cttgacgtcg gttaccgata tccatatggc ggccgcatcg atctcgagcc
      661 gcggactagt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
      721 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
      781 caatgtatct tatcatgtct tacgtagata agtagcatgg cgggttaatc attaactaca
      841 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
      901 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
      961 gagcgcgcag agagggacag atctgcagat ctgctggcgt aatagcgaag aggcccgcac
     1021 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt aagcgttaat
     1081 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     1141 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     1201 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     1261 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     1321 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     1381 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     1441 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     1501 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1561 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1621 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1681 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1741 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1801 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1861 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1921 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1981 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2041 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2101 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2161 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     2221 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2281 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2341 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2401 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2461 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2521 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2581 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2761 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2821 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2881 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2941 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3001 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3061 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3241 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3301 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3361 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3421 cgcagcgagt cagtgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     3481 cctggagggg tggagtcgtg acgtgaatta cgtcataggg ttagggaggt cctggatcga
     3541 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3601 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3661 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3721 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatct
     3781 ctagtcaagg cactatacat caaatattcc ttattaaccc ctttacaaat taaaaagcta
     3841 aaggtacaca atttttgagc atagttatta atagcagaca ctctatgcct gtgtggagta
     3901 agaaaaaaca gtatgttatg attataactg ttatgcctac ttataaaggt tacagaatat
     3961 ttttccataa ttttcttgta tagcagtgca gctttttcct ttgtggtgta aatagcaaag
     4021 caagcaagag ttctattact aaacacagca tgactcaaaa aacttagcaa ttctgaagga
     4081 aagtccttgg ggtcttctac ctttctcttc ttttttggag gagtagaatg ttgagagtca
     4141 gcagtagcct catcatcact agatggcatt tcttctgagc aaaacaggtt ttcctcatta
     4201 aaggcattcc accactgctc ccattcatca gttccatagg ttggaatcta aaatacacaa
     4261 acaattagaa tcagtagttt aacacattat acacttaaaa attttatatt taccttagag
     4321 ctttaaatct ctgtaggtag tttgtccaat tatgtcacac cacagaagta aggttccttc
     4381 acaaagatcc ggaccaaagc ggccatcgtg cctccccact cctgcagttc gggggcatgg
     4441 atgcgcggat agccgctgct ggtttcctgg atgccgacgg atttgcactg ccggtagaac
     4501 tccgcgaggt cgtccagcct caggcagcag ctgaaccaac tcgcgagggg atcgagcccg
     4561 gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc
     4621 cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt
     4681 gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga
     4741 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     4801 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     4861 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     4921 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat
     4981 cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     5041 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     5101 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     5161 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
     5221 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     5281 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     5341 cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     5401 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     5461 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     5521 caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc ctgcgccatc
     5581 agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag
     5641 agggcgcccc agctggcaat tccggttcgc ttgctgtcca taaaaccgcc cagtctagct
     5701 atcgccatgt aagcccactg caagctacct gctttctctt tgcgcttgcg ttttcccttg
     5761 tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt
     5821 tctacgtgtt ccgcttcctt tagcagccct tgcgccctga gtgcttgcgg cagcgtgaag
     5881 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     5941 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     6001 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     6061 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     6121 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     6181 gcctggggac tttccacacc ctaactgaca cacattccac agccggatcc aaggtcgggc
     6241 aggaa
//
