LOCUS       sequence-311767-        6265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impacts 1 uracil in a poly-uracil tract in 
            conjunctiuon with an immediately upstream hairpin 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 6265)
  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 6265)
  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)_HairpinDistance(0)nts_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        45..293
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     45..65
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     216..235
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    735..856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   905..1034
                     /label=AAV2 ITR
     rep_origin      complement(1128..1583)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1265..1286)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1477..1496
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1610..1714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1715..1783,1784..2575)
                     /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(1933..1952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2746..3334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3235..3254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3603..3737
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4162..4182)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4312..4377
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4797..5591)
                     /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(4908..4927)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5518..5537
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5948..6265,1..12))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5962..6097
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     join(6262..6265,1..17)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 taactgacac acattccaca gccggatcca aggtcgggca ggaagagggc ctatttccca
       61 tgattccttc atatttgcat atacgataca aggctgttag agagataatt agaattaatt
      121 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
      181 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
      241 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgtgctcg
      301 cttcggcagc acatatacta gttggcggtt gcggctttct gctgcaatct cgacggccat
      361 cagtcgccgg tcccaagccc ggataaaatg ggagggggcg ggaaaccgcc taaccatgcc
      421 gactgatggc aggcccggat agctcagtcg gtagagcagc ggccgcgcga ttcgaggaag
      481 gaggtctgag gaggtcactg aatcgcgcgg ccgcgggtcc agggttcaag tccctgttcg
      541 ggcctgccat cagtcggcgt ggactgtaga acactgccaa tgccggtccc aagcccggat
      601 aaaagtggag ggtacagtcc acgctctaga gcggacttcg gtccgctttt tactaggacc
      661 tgcaggcatg caagcttgac gtcggttacc gatatccata tggcggccgc atcgatctcg
      721 agccgcggac tagtaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
      781 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
      841 tcatcaatgt atcttatcat gtcttacgta gataagtagc atggcgggtt aatcattaac
      901 tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
      961 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     1021 gagcgagcgc gcagagaggg acagatctgc agatctgctg gcgtaatagc gaagaggccc
     1081 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt
     1141 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     1201 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     1261 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     1321 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     1381 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     1441 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     1501 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     1561 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1621 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1681 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     1741 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1801 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1861 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1921 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     1981 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2041 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2101 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2161 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2221 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     2281 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2341 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2401 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     2461 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2521 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     2581 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     2641 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     2701 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2761 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2821 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2881 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2941 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3001 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3061 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3121 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3181 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3241 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3301 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3361 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3421 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     3481 cgagcgcagc gagtcagtga gcgagcgcgc agagagggag tggccaactc catcactagg
     3541 ggttcctgga ggggtggagt cgtgacgtga attacgtcat agggttaggg aggtcctgga
     3601 tcgatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3661 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3721 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3781 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     3841 atctctagtc aaggcactat acatcaaata ttccttatta acccctttac aaattaaaaa
     3901 gctaaaggta cacaattttt gagcatagtt attaatagca gacactctat gcctgtgtgg
     3961 agtaagaaaa aacagtatgt tatgattata actgttatgc ctacttataa aggttacaga
     4021 atatttttcc ataattttct tgtatagcag tgcagctttt tcctttgtgg tgtaaatagc
     4081 aaagcaagca agagttctat tactaaacac agcatgactc aaaaaactta gcaattctga
     4141 aggaaagtcc ttggggtctt ctacctttct cttctttttt ggaggagtag aatgttgaga
     4201 gtcagcagta gcctcatcat cactagatgg catttcttct gagcaaaaca ggttttcctc
     4261 attaaaggca ttccaccact gctcccattc atcagttcca taggttggaa tctaaaatac
     4321 acaaacaatt agaatcagta gtttaacaca ttatacactt aaaaatttta tatttacctt
     4381 agagctttaa atctctgtag gtagtttgtc caattatgtc acaccacaga agtaaggttc
     4441 cttcacaaag atccggacca aagcggccat cgtgcctccc cactcctgca gttcgggggc
     4501 atggatgcgc ggatagccgc tgctggtttc ctggatgccg acggatttgc actgccggta
     4561 gaactccgcg aggtcgtcca gcctcaggca gcagctgaac caactcgcga ggggatcgag
     4621 cccggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc
     4681 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc
     4741 tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag
     4801 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     4861 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     4921 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     4981 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     5041 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc
     5101 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     5161 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     5221 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     5281 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     5341 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     5401 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     5461 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     5521 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     5581 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatcttga tcccctgcgc
     5641 catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta
     5701 ccagagggcg ccccagctgg caattccggt tcgcttgctg tccataaaac cgcccagtct
     5761 agctatcgcc atgtaagccc actgcaagct acctgctttc tctttgcgct tgcgttttcc
     5821 cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg
     5881 gctttctacg tgttccgctt cctttagcag cccttgcgcc ctgagtgctt gcggcagcgt
     5941 gaagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     6001 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     6061 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     6121 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     6181 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     6241 gggagcctgg ggactttcca caccc
//
