LOCUS       sequence-311748-        6244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 3 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 6244)
  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 6244)
  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(3)
FEATURES             Location/Qualifiers
     source          1..6244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..83
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    562..683
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   732..861
                     /label=AAV2 ITR
     rep_origin      complement(955..1410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1092..1113)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1304..1323
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1437..1541
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1542..1610,1611..2402)
                     /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(1760..1779)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2573..3161
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3062..3081
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3430..3564
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3989..4009)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4139..4204
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4624..5418)
                     /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(4735..4754)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5345..5364
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5775..6104)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5789..5924
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6089..6109
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6137..6244,1..141)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6137..6157
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
       61 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
      121 ttgtggaaag gacgaaacac cgtgctcgct tcggcagcac atatactagt tggctttcga
      181 cggccatcag tcgccggtcc caagcccgga taaaatggga gggggcggga aaccgcctaa
      241 ccatgccgac tgatggcagg cccggatagc tcagtcggta gagcagcggc cgcgcgattc
      301 gaggaaggag gtctgaggag gtcactgaat cgcgcggccg cgggtccagg gttcaagtcc
      361 ctgttcgggc ctgccatcag tcggcgtgga ctgtagaaca ctgccaatgc cggtcccaag
      421 cccggataaa agtggagggt acagtccacg ctctagagcg gacttcggtc cgctttttac
      481 taggacctgc aggcatgcaa gcttgacgtc ggttaccgat atccatatgg cggccgcatc
      541 gatctcgagc cgcggactag taacttgttt attgcagctt ataatggtta caaataaagc
      601 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
      661 tccaaactca tcaatgtatc ttatcatgtc ttacgtagat aagtagcatg gcgggttaat
      721 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
      781 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
      841 agtgagcgag cgagcgcgca gagagggaca gatctgcaga tctgctggcg taatagcgaa
      901 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggaaattg
      961 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     1021 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     1081 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     1141 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     1201 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     1261 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     1321 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     1381 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     1441 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1501 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1561 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1621 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     1681 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     1741 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     1801 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     1861 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     1921 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     1981 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2041 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2101 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2161 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2221 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2281 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2341 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2401 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2461 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2521 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2581 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2641 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2701 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2761 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2821 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2881 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2941 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3001 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3061 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3121 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3181 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3241 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3301 gaacgaccga gcgcagcgag tcagtgagcg agcgcgcaga gagggagtgg ccaactccat
     3361 cactaggggt tcctggaggg gtggagtcgt gacgtgaatt acgtcatagg gttagggagg
     3421 tcctggatcg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3481 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3541 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3601 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
     3661 gattatgatc tctagtcaag gcactataca tcaaatattc cttattaacc cctttacaaa
     3721 ttaaaaagct aaaggtacac aatttttgag catagttatt aatagcagac actctatgcc
     3781 tgtgtggagt aagaaaaaac agtatgttat gattataact gttatgccta cttataaagg
     3841 ttacagaata tttttccata attttcttgt atagcagtgc agctttttcc tttgtggtgt
     3901 aaatagcaaa gcaagcaaga gttctattac taaacacagc atgactcaaa aaacttagca
     3961 attctgaagg aaagtccttg gggtcttcta cctttctctt cttttttgga ggagtagaat
     4021 gttgagagtc agcagtagcc tcatcatcac tagatggcat ttcttctgag caaaacaggt
     4081 tttcctcatt aaaggcattc caccactgct cccattcatc agttccatag gttggaatct
     4141 aaaatacaca aacaattaga atcagtagtt taacacatta tacacttaaa aattttatat
     4201 ttaccttaga gctttaaatc tctgtaggta gtttgtccaa ttatgtcaca ccacagaagt
     4261 aaggttcctt cacaaagatc cggaccaaag cggccatcgt gcctccccac tcctgcagtt
     4321 cgggggcatg gatgcgcgga tagccgctgc tggtttcctg gatgccgacg gatttgcact
     4381 gccggtagaa ctccgcgagg tcgtccagcc tcaggcagca gctgaaccaa ctcgcgaggg
     4441 gatcgagccc ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     4501 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     4561 cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc
     4621 cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
     4681 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
     4741 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
     4801 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
     4861 cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg
     4921 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
     4981 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
     5041 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
     5101 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
     5161 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
     5221 ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
     5281 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
     5341 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
     5401 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc
     5461 cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc
     5521 aaccttacca gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc
     5581 ccagtctagc tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc
     5641 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
     5701 cggactggct ttctacgtgt tccgcttcct ttagcagccc ttgcgccctg agtgcttgcg
     5761 gcagcgtgaa gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     5821 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     5881 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     5941 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     6001 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     6061 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagccggatc
     6121 caaggtcggg caggaagagg gcctatttcc catgattcct tcatatttgc atatacgata
     6181 caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca
     6241 aaat
//
