LOCUS       sequence-311759-        6276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impacts 4 uracils in a poly-uracil tract in 
            conjunctiuon with a hairpin 19 nts upstream 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 6276)
  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 6276)
  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-3_PolyU(4)_HairpinDistance(19)nts_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..111
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    622..743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   792..921
                     /label=AAV2 ITR
     rep_origin      complement(1015..1470)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1152..1173)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1364..1383
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1497..1601
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1602..1670,1671..2462)
                     /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(1820..1839)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2633..3221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3122..3141
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3490..3624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4049..4069)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4199..4264
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4684..5478)
                     /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(4795..4814)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5405..5424
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5835..6164)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5849..5984
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6149..6169
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6197..6276,1..169)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6197..6217
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
       61 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
      121 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg tgctcgcttc
      181 ggcagcacat aggttgcggc tttctgctgc aatcaacaag cagccaactc caattttcga
      241 cggccatcag tcgccggtcc caagcccgga taaaatggga gggggcggga aaccgcctaa
      301 ccatgccgac tgatggcagg cccggatagc tcagtcggta gagcagcggc cgcgcgattc
      361 gaggaaggag gtctgaggag gtcactgaat cgcgcggccg cgggtccagg gttcaagtcc
      421 ctgttcgggc ctgccatcag tcggcgtgga ctgtagaaca ctgccaatgc cggtcccaag
      481 cccggataaa agtggagggt acagtccacg ctctagagcg gacttcggtc cgctttttac
      541 taggacctgc aggcatgcaa gcttgacgtc ggttaccgat atccatatgg cggccgcatc
      601 gatctcgagc cgcggactag taacttgttt attgcagctt ataatggtta caaataaagc
      661 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
      721 tccaaactca tcaatgtatc ttatcatgtc ttacgtagat aagtagcatg gcgggttaat
      781 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
      841 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
      901 agtgagcgag cgagcgcgca gagagggaca gatctgcaga tctgctggcg taatagcgaa
      961 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggaaattg
     1021 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     1081 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     1141 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     1201 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     1261 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     1321 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     1381 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     1441 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     1501 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1561 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1621 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1681 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     1741 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     1801 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     1861 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     1921 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     1981 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2041 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2101 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2161 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2221 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2281 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2341 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2401 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2461 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2521 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2581 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2641 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2701 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2761 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2821 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2881 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2941 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3001 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3061 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3121 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3181 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3241 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3301 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3361 gaacgaccga gcgcagcgag tcagtgagcg agcgcgcaga gagggagtgg ccaactccat
     3421 cactaggggt tcctggaggg gtggagtcgt gacgtgaatt acgtcatagg gttagggagg
     3481 tcctggatcg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3541 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3601 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3661 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     3721 gattatgatc tctagtcaag gcactataca tcaaatattc cttattaacc cctttacaaa
     3781 ttaaaaagct aaaggtacac aatttttgag catagttatt aatagcagac actctatgcc
     3841 tgtgtggagt aagaaaaaac agtatgttat gattataact gttatgccta cttataaagg
     3901 ttacagaata tttttccata attttcttgt atagcagtgc agctttttcc tttgtggtgt
     3961 aaatagcaaa gcaagcaaga gttctattac taaacacagc atgactcaaa aaacttagca
     4021 attctgaagg aaagtccttg gggtcttcta cctttctctt cttttttgga ggagtagaat
     4081 gttgagagtc agcagtagcc tcatcatcac tagatggcat ttcttctgag caaaacaggt
     4141 tttcctcatt aaaggcattc caccactgct cccattcatc agttccatag gttggaatct
     4201 aaaatacaca aacaattaga atcagtagtt taacacatta tacacttaaa aattttatat
     4261 ttaccttaga gctttaaatc tctgtaggta gtttgtccaa ttatgtcaca ccacagaagt
     4321 aaggttcctt cacaaagatc cggaccaaag cggccatcgt gcctccccac tcctgcagtt
     4381 cgggggcatg gatgcgcgga tagccgctgc tggtttcctg gatgccgacg gatttgcact
     4441 gccggtagaa ctccgcgagg tcgtccagcc tcaggcagca gctgaaccaa ctcgcgaggg
     4501 gatcgagccc ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     4561 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     4621 cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc
     4681 cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     4741 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     4801 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     4861 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     4921 cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg
     4981 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     5041 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     5101 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     5161 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     5221 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     5281 ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     5341 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     5401 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     5461 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc
     5521 cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc
     5581 aaccttacca gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc
     5641 ccagtctagc tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc
     5701 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
     5761 cggactggct ttctacgtgt tccgcttcct ttagcagccc ttgcgccctg agtgcttgcg
     5821 gcagcgtgaa gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     5881 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     5941 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     6001 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     6061 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     6121 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagccggatc
     6181 caaggtcggg caggaagagg gcctatttcc catgattcct tcatatttgc atatacgata
     6241 caaggctgtt agagagataa ttagaattaa tttgac
//
