LOCUS       sequence-311781-        6242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 1 uracil 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 6242)
  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 6242)
  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(1)
FEATURES             Location/Qualifiers
     source          1..6242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   102..231
                     /label=AAV2 ITR
     rep_origin      complement(325..780)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(462..483)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     674..693
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        807..911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(912..980,981..1772)
                     /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(1130..1149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1943..2531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2432..2451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2800..2934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3359..3379)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          3509..3574
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(3994..4788)
                     /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(4105..4124)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4715..4734
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5145..5474)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5159..5294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5459..5479
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5507..5755
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5507..5527
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5678..5697
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    join(6174..6242,1..53)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
       61 ttacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga
      121 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
      181 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gagagggaca
      241 gatctgcaga tctgctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
      301 ttgcgcagcc tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt
      361 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
      421 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
      481 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
      541 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
      601 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
      661 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
      721 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
      781 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
      841 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      901 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      961 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1021 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1081 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1141 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1201 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1261 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1321 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1381 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1441 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1501 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1561 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1621 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     1681 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     1741 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1801 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1861 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1921 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1981 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2041 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2101 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2161 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2221 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2281 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2341 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2401 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2461 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2521 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2581 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     2641 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2701 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctggaggg gtggagtcgt
     2761 gacgtgaatt acgtcatagg gttagggagg tcctggatcg atccagacat gataagatac
     2821 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2881 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2941 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     3001 aagtaaaacc tctacaaatg tggtatggct gattatgatc tctagtcaag gcactataca
     3061 tcaaatattc cttattaacc cctttacaaa ttaaaaagct aaaggtacac aatttttgag
     3121 catagttatt aatagcagac actctatgcc tgtgtggagt aagaaaaaac agtatgttat
     3181 gattataact gttatgccta cttataaagg ttacagaata tttttccata attttcttgt
     3241 atagcagtgc agctttttcc tttgtggtgt aaatagcaaa gcaagcaaga gttctattac
     3301 taaacacagc atgactcaaa aaacttagca attctgaagg aaagtccttg gggtcttcta
     3361 cctttctctt cttttttgga ggagtagaat gttgagagtc agcagtagcc tcatcatcac
     3421 tagatggcat ttcttctgag caaaacaggt tttcctcatt aaaggcattc caccactgct
     3481 cccattcatc agttccatag gttggaatct aaaatacaca aacaattaga atcagtagtt
     3541 taacacatta tacacttaaa aattttatat ttaccttaga gctttaaatc tctgtaggta
     3601 gtttgtccaa ttatgtcaca ccacagaagt aaggttcctt cacaaagatc cggaccaaag
     3661 cggccatcgt gcctccccac tcctgcagtt cgggggcatg gatgcgcgga tagccgctgc
     3721 tggtttcctg gatgccgacg gatttgcact gccggtagaa ctccgcgagg tcgtccagcc
     3781 tcaggcagca gctgaaccaa ctcgcgaggg gatcgagccc ggggtgggcg aagaactcca
     3841 gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc
     3901 ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg
     3961 cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata
     4021 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
     4081 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
     4141 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
     4201 gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg
     4261 cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
     4321 atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc
     4381 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
     4441 catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac
     4501 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
     4561 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag
     4621 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa
     4681 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
     4741 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc
     4801 tcatcctgtc tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa
     4861 agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa
     4921 ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact
     4981 gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct
     5041 gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtgt tccgcttcct
     5101 ttagcagccc ttgcgccctg agtgcttgcg gcagcgtgaa gctttttgca aaagcctagg
     5161 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     5221 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     5281 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     5341 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     5401 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     5461 cctaactgac acacattcca cagccggatc caaggtcggg caggaagagg gcctatttcc
     5521 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa
     5581 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
     5641 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
     5701 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgtgct
     5761 cgcttcggca gcacatatac tagttggctc gacggccatc agtcgccggt cccaagcccg
     5821 gataaaatgg gagggggcgg gaaaccgcct aaccatgccg actgatggca ggcccggata
     5881 gctcagtcgg tagagcagcg gccgcgcgat tcgaggaagg aggtctgagg aggtcactga
     5941 atcgcgcggc cgcgggtcca gggttcaagt ccctgttcgg gcctgccatc agtcggcgtg
     6001 gactgtagaa cactgccaat gccggtccca agcccggata aaagtggagg gtacagtcca
     6061 cgctctagag cggacttcgg tccgcttttt actaggacct gcaggcatgc aagcttgacg
     6121 tcggttaccg atatccatat ggcggccgca tcgatctcga gccgcggact agtaacttgt
     6181 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6241 ca
//
