LOCUS       sequence-311768-        6249 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 6249)
  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 6249)
  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-2_PolyU(4)_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    112..233
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   282..411
                     /label=AAV2 ITR
     rep_origin      complement(505..960)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(642..663)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     854..873
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        987..1091
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1092..1160,1161..1952)
                     /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(1310..1329)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2123..2711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2612..2631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2980..3114
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3539..3559)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          3689..3754
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4174..4968)
                     /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(4285..4304)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4895..4914
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5325..5654)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5339..5474
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5639..5659
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5687..5935
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5687..5707
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5858..5877
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 ctctagagcg gacttcggtc cgctttttac taggacctgc aggcatgcaa gcttgacgtc
       61 ggttaccgat atccatatgg cggccgcatc gatctcgagc cgcggactag taacttgttt
      121 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
      181 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
      241 ttacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga
      301 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
      361 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gagagggaca
      421 gatctgcaga tctgctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
      481 ttgcgcagcc tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt
      541 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
      601 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
      661 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
      721 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
      781 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
      841 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
      901 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
      961 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1021 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1081 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1141 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1201 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1261 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1321 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1381 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1441 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1501 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1561 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1621 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1681 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1741 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1801 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     1861 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     1921 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1981 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2041 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2101 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2161 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2221 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2281 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2341 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2401 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2461 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2521 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2581 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2641 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2701 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2761 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     2821 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2881 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctggaggg gtggagtcgt
     2941 gacgtgaatt acgtcatagg gttagggagg tcctggatcg atccagacat gataagatac
     3001 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3061 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3121 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     3181 aagtaaaacc tctacaaatg tggtatggct gattatgatc tctagtcaag gcactataca
     3241 tcaaatattc cttattaacc cctttacaaa ttaaaaagct aaaggtacac aatttttgag
     3301 catagttatt aatagcagac actctatgcc tgtgtggagt aagaaaaaac agtatgttat
     3361 gattataact gttatgccta cttataaagg ttacagaata tttttccata attttcttgt
     3421 atagcagtgc agctttttcc tttgtggtgt aaatagcaaa gcaagcaaga gttctattac
     3481 taaacacagc atgactcaaa aaacttagca attctgaagg aaagtccttg gggtcttcta
     3541 cctttctctt cttttttgga ggagtagaat gttgagagtc agcagtagcc tcatcatcac
     3601 tagatggcat ttcttctgag caaaacaggt tttcctcatt aaaggcattc caccactgct
     3661 cccattcatc agttccatag gttggaatct aaaatacaca aacaattaga atcagtagtt
     3721 taacacatta tacacttaaa aattttatat ttaccttaga gctttaaatc tctgtaggta
     3781 gtttgtccaa ttatgtcaca ccacagaagt aaggttcctt cacaaagatc cggaccaaag
     3841 cggccatcgt gcctccccac tcctgcagtt cgggggcatg gatgcgcgga tagccgctgc
     3901 tggtttcctg gatgccgacg gatttgcact gccggtagaa ctccgcgagg tcgtccagcc
     3961 tcaggcagca gctgaaccaa ctcgcgaggg gatcgagccc ggggtgggcg aagaactcca
     4021 gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc
     4081 ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg
     4141 cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata
     4201 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
     4261 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
     4321 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
     4381 gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg
     4441 cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
     4501 atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc
     4561 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
     4621 catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac
     4681 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
     4741 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag
     4801 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa
     4861 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
     4921 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc
     4981 tcatcctgtc tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa
     5041 agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa
     5101 ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact
     5161 gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct
     5221 gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtgt tccgcttcct
     5281 ttagcagccc ttgcgccctg agtgcttgcg gcagcgtgaa gctttttgca aaagcctagg
     5341 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     5401 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     5461 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     5521 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     5581 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     5641 cctaactgac acacattcca cagccggatc caaggtcggg caggaagagg gcctatttcc
     5701 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa
     5761 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
     5821 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
     5881 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgtgct
     5941 cgcttcggca gcacatatac tagtgattga tgttttcgac ggccatcagt cgccggtccc
     6001 aagcccggat aaaatgggag ggggcgggaa accgcctaac catgccgact gatggcaggc
     6061 ccggatagct cagtcggtag agcagcggcc gcgcgattcg aggaaggagg tctgaggagg
     6121 tcactgaatc gcgcggccgc gggtccaggg ttcaagtccc tgttcgggcc tgccatcagt
     6181 cggcgtggac tgtagaacac tgccaatgcc ggtcccaagc ccggataaaa gtggagggta
     6241 cagtccacg
//
