LOCUS       sequence-311734-        6246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 5 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 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-1_PolyU(5)_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    431..552
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   601..730
                     /label=AAV2 ITR
     rep_origin      complement(824..1279)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(961..982)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1173..1192
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1306..1410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1411..1479,1480..2271)
                     /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(1629..1648)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2442..3030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2931..2950
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3299..3433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3858..3878)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4008..4073
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4493..5287)
                     /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(4604..4623)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5214..5233
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5644..5973)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5658..5793
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5958..5978
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6006..6246,1..8)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6006..6026
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6177..6196
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gaaacaccgt gctcgcttcg gcagcacata tactagttgg ctttttcgac ggccatcagt
       61 cgccggtccc aagcccggat aaaatgggag ggggcgggaa accgcctaac catgccgact
      121 gatggcaggc ccggatagct cagtcggtag agcagcggcc gcgcgattcg aggaaggagg
      181 tctgaggagg tcactgaatc gcgcggccgc gggtccaggg ttcaagtccc tgttcgggcc
      241 tgccatcagt cggcgtggac tgtagaacac tgccaatgcc ggtcccaagc ccggataaaa
      301 gtggagggta cagtccacgc tctagagcgg acttcggtcc gctttttact aggacctgca
      361 ggcatgcaag cttgacgtcg gttaccgata tccatatggc ggccgcatcg atctcgagcc
      421 gcggactagt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
      481 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
      541 caatgtatct tatcatgtct tacgtagata agtagcatgg cgggttaatc attaactaca
      601 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
      661 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
      721 gagcgcgcag agagggacag atctgcagat ctgctggcgt aatagcgaag aggcccgcac
      781 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt aagcgttaat
      841 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
      901 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
      961 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     1021 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     1081 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     1141 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     1201 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     1261 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1321 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1381 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1441 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1501 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1561 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1621 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1681 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1741 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1801 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1861 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1921 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1981 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2041 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2101 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2161 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2221 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2281 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2341 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2401 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2461 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2521 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2581 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2641 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2701 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2761 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2821 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2881 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2941 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3001 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3061 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3121 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3181 cgcagcgagt cagtgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     3241 cctggagggg tggagtcgtg acgtgaatta cgtcataggg ttagggaggt cctggatcga
     3301 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3361 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3421 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     3481 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatct
     3541 ctagtcaagg cactatacat caaatattcc ttattaaccc ctttacaaat taaaaagcta
     3601 aaggtacaca atttttgagc atagttatta atagcagaca ctctatgcct gtgtggagta
     3661 agaaaaaaca gtatgttatg attataactg ttatgcctac ttataaaggt tacagaatat
     3721 ttttccataa ttttcttgta tagcagtgca gctttttcct ttgtggtgta aatagcaaag
     3781 caagcaagag ttctattact aaacacagca tgactcaaaa aacttagcaa ttctgaagga
     3841 aagtccttgg ggtcttctac ctttctcttc ttttttggag gagtagaatg ttgagagtca
     3901 gcagtagcct catcatcact agatggcatt tcttctgagc aaaacaggtt ttcctcatta
     3961 aaggcattcc accactgctc ccattcatca gttccatagg ttggaatcta aaatacacaa
     4021 acaattagaa tcagtagttt aacacattat acacttaaaa attttatatt taccttagag
     4081 ctttaaatct ctgtaggtag tttgtccaat tatgtcacac cacagaagta aggttccttc
     4141 acaaagatcc ggaccaaagc ggccatcgtg cctccccact cctgcagttc gggggcatgg
     4201 atgcgcggat agccgctgct ggtttcctgg atgccgacgg atttgcactg ccggtagaac
     4261 tccgcgaggt cgtccagcct caggcagcag ctgaaccaac tcgcgagggg atcgagcccg
     4321 gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc
     4381 cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt
     4441 gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga
     4501 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     4561 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     4621 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     4681 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat
     4741 cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     4801 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     4861 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     4921 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
     4981 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     5041 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     5101 cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     5161 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     5221 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     5281 caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc ctgcgccatc
     5341 agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag
     5401 agggcgcccc agctggcaat tccggttcgc ttgctgtcca taaaaccgcc cagtctagct
     5461 atcgccatgt aagcccactg caagctacct gctttctctt tgcgcttgcg ttttcccttg
     5521 tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt
     5581 tctacgtgtt ccgcttcctt tagcagccct tgcgccctga gtgcttgcgg cagcgtgaag
     5641 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     5701 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     5761 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     5821 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     5881 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     5941 gcctggggac tttccacacc ctaactgaca cacattccac agccggatcc aaggtcgggc
     6001 aggaagaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     6061 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     6121 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     6181 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     6241 aaggac
//
