LOCUS       sequence-311745-        6269 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 6269)
  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 6269)
  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(1)_HairpinDistance(0)nts_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..83
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    587..708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   757..886
                     /label=AAV2 ITR
     rep_origin      complement(980..1435)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1117..1138)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1329..1348
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1462..1566
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1567..1635,1636..2427)
                     /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(1785..1804)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2598..3186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3087..3106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3455..3589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4014..4034)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4164..4229
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4649..5443)
                     /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
                     GLAPAELFARLKARMPDDEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4760..4779)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5370..5389
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5800..6129)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5814..5949
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6114..6134
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6162..6269,1..141)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6162..6182
                     /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 gattgatggg
      181 ttgcggcttt ctgctgcaat ctcgacggcc atcagtcgcc ggtcccaagc ccggataaaa
      241 tgggaggggg cgggaaaccg cctaaccatg ccgactgatg gcaggcccgg atagctcagt
      301 cggtagagca gcggccgcgc gattcgagga aggaggtctg aggaggtcac tgaatcgcgc
      361 ggccgcgggt ccagggttca agtccctgtt cgggcctgcc atcagtcggc gtggactgta
      421 gaacactgcc aatgccggtc ccaagcccgg ataaaagtgg agggtacagt ccacgctcta
      481 gagcggactt cggtccgctt tttactagga cctgcaggca tgcaagcttg acgtcggtta
      541 ccgatatcca tatggcggcc gcatcgatct cgagccgcgg actagtaact tgtttattgc
      601 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      661 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtcttacg
      721 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
      781 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
      841 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagagag ggacagatct
      901 gcagatctgc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
      961 cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     1021 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     1081 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     1141 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     1201 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     1261 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     1321 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     1381 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     1441 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1501 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1561 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1621 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1681 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1741 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1801 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1861 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1921 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1981 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     2041 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     2101 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     2161 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2221 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2281 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2341 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2401 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2461 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2521 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2581 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2641 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2701 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2761 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2821 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2881 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2941 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3001 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3061 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3121 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3181 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3241 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3301 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgagcgc
     3361 gcagagaggg agtggccaac tccatcacta ggggttcctg gaggggtgga gtcgtgacgt
     3421 gaattacgtc atagggttag ggaggtcctg gatcgatcca gacatgataa gatacattga
     3481 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3541 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3601 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     3661 aaacctctac aaatgtggta tggctgatta tgatctctag tcaaggcact atacatcaaa
     3721 tattccttat taaccccttt acaaattaaa aagctaaagg tacacaattt ttgagcatag
     3781 ttattaatag cagacactct atgcctgtgt ggagtaagaa aaaacagtat gttatgatta
     3841 taactgttat gcctacttat aaaggttaca gaatattttt ccataatttt cttgtatagc
     3901 agtgcagctt tttcctttgt ggtgtaaata gcaaagcaag caagagttct attactaaac
     3961 acagcatgac tcaaaaaact tagcaattct gaaggaaagt ccttggggtc ttctaccttt
     4021 ctcttctttt ttggaggagt agaatgttga gagtcagcag tagcctcatc atcactagat
     4081 ggcatttctt ctgagcaaaa caggttttcc tcattaaagg cattccacca ctgctcccat
     4141 tcatcagttc cataggttgg aatctaaaat acacaaacaa ttagaatcag tagtttaaca
     4201 cattatacac ttaaaaattt tatatttacc ttagagcttt aaatctctgt aggtagtttg
     4261 tccaattatg tcacaccaca gaagtaaggt tccttcacaa agatccggac caaagcggcc
     4321 atcgtgcctc cccactcctg cagttcgggg gcatggatgc gcggatagcc gctgctggtt
     4381 tcctggatgc cgacggattt gcactgccgg tagaactccg cgaggtcgtc cagcctcagg
     4441 cagcagctga accaactcgc gaggggatcg agcccggggt gggcgaagaa ctccagcatg
     4501 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     4561 ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg
     4621 tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     4681 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     4741 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     4801 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     4861 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gtcgtcgggc atgcgcgcct
     4921 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     4981 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     5041 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     5101 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     5161 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     5221 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac
     5281 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     5341 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     5401 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     5461 ctgtctcttg atcagatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca
     5521 tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg
     5581 gttcgcttgc tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag
     5641 ctacctgctt tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat
     5701 tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtgttccgc ttcctttagc
     5761 agcccttgcg ccctgagtgc ttgcggcagc gtgaagcttt ttgcaaaagc ctaggcctcc
     5821 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     5881 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     5941 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     6001 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     6061 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     6121 ctgacacaca ttccacagcc ggatccaagg tcgggcagga agagggccta tttcccatga
     6181 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga
     6241 ctgtaaacac aaagatatta gtacaaaat
//
