LOCUS       sequence-311732-        6276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impacts 4 uracils in a poly-uracil tract in 
            conjunctiuon with a hairpin 10 nts upstream 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 6276)
  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 6276)
  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-3_PolyU(4)_HairpinDistance(10)nts_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   7..136
                     /label=AAV2 ITR
     rep_origin      complement(230..685)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(367..388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     579..598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        712..816
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(817..885,886..1677)
                     /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(1035..1054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1848..2436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2337..2356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2705..2839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3264..3284)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          3414..3479
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(3899..4693)
                     /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(4010..4029)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4620..4639
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5050..5379)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5064..5199
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5364..5384
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5412..5660
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5412..5432
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5583..5602
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    6113..6234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
       61 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
      121 gcgagcgagc gcgcagagag ggacagatct gcagatctgc tggcgtaata gcgaagaggc
      181 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga aattgtaagc
      241 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
      301 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
      361 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
      421 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
      481 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
      541 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
      601 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
      661 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
      721 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
      781 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
      841 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
      901 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
      961 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1021 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     1081 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1141 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1201 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     1261 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     1321 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     1381 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     1441 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     1501 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     1561 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     1621 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     1681 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     1741 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     1801 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1861 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1921 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1981 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2041 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2101 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2161 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2221 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2281 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2341 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2401 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2461 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2521 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     2581 accgagcgca gcgagtcagt gagcgagcgc gcagagaggg agtggccaac tccatcacta
     2641 ggggttcctg gaggggtgga gtcgtgacgt gaattacgtc atagggttag ggaggtcctg
     2701 gatcgatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2761 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2821 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2881 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     2941 tgatctctag tcaaggcact atacatcaaa tattccttat taaccccttt acaaattaaa
     3001 aagctaaagg tacacaattt ttgagcatag ttattaatag cagacactct atgcctgtgt
     3061 ggagtaagaa aaaacagtat gttatgatta taactgttat gcctacttat aaaggttaca
     3121 gaatattttt ccataatttt cttgtatagc agtgcagctt tttcctttgt ggtgtaaata
     3181 gcaaagcaag caagagttct attactaaac acagcatgac tcaaaaaact tagcaattct
     3241 gaaggaaagt ccttggggtc ttctaccttt ctcttctttt ttggaggagt agaatgttga
     3301 gagtcagcag tagcctcatc atcactagat ggcatttctt ctgagcaaaa caggttttcc
     3361 tcattaaagg cattccacca ctgctcccat tcatcagttc cataggttgg aatctaaaat
     3421 acacaaacaa ttagaatcag tagtttaaca cattatacac ttaaaaattt tatatttacc
     3481 ttagagcttt aaatctctgt aggtagtttg tccaattatg tcacaccaca gaagtaaggt
     3541 tccttcacaa agatccggac caaagcggcc atcgtgcctc cccactcctg cagttcgggg
     3601 gcatggatgc gcggatagcc gctgctggtt tcctggatgc cgacggattt gcactgccgg
     3661 tagaactccg cgaggtcgtc cagcctcagg cagcagctga accaactcgc gaggggatcg
     3721 agcccggggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
     3781 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
     3841 tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc
     3901 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     3961 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     4021 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
     4081 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     4141 cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga
     4201 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     4261 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     4321 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     4381 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     4441 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     4501 ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     4561 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     4621 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     4681 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatctt gatcccctgc
     4741 gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct
     4801 taccagaggg cgccccagct ggcaattccg gttcgcttgc tgtccataaa accgcccagt
     4861 ctagctatcg ccatgtaagc ccactgcaag ctacctgctt tctctttgcg cttgcgtttt
     4921 cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac
     4981 tggctttcta cgtgttccgc ttcctttagc agcccttgcg ccctgagtgc ttgcggcagc
     5041 gtgaagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     5101 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     5161 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     5221 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     5281 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5341 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagcc ggatccaagg
     5401 tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     5461 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
     5521 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     5581 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     5641 tgtggaaagg acgaaacacc gtgctcgctt cggcagcaca tatactagca gggttgcggc
     5701 tttctgctgc aatcccaact ccaattttcg acggccatca gtcgccggtc ccaagcccgg
     5761 ataaaatggg agggggcggg aaaccgccta accatgccga ctgatggcag gcccggatag
     5821 ctcagtcggt agagcagcgg ccgcgcgatt cgaggaagga ggtctgagga ggtcactgaa
     5881 tcgcgcggcc gcgggtccag ggttcaagtc cctgttcggg cctgccatca gtcggcgtgg
     5941 actgtagaac actgccaatg ccggtcccaa gcccggataa aagtggaggg tacagtccac
     6001 gctctagagc ggacttcggt ccgcttttta ctaggacctg caggcatgca agcttgacgt
     6061 cggttaccga tatccatatg gcggccgcat cgatctcgag ccgcggacta gtaacttgtt
     6121 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6181 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6241 cttacgtaga taagtagcat ggcgggttaa tcatta
//
