LOCUS       sequence-311735-        6252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 7 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 6252)
  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 6252)
  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(7)_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    437..558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   607..736
                     /label=AAV2 ITR
     rep_origin      complement(830..1285)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(967..988)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1179..1198
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1312..1416
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1417..1485,1486..2277)
                     /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(1635..1654)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2448..3036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2937..2956
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3305..3439
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3864..3884)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4014..4079
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4499..5293)
                     /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(4610..4629)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5220..5239
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5650..5979)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5664..5799
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5964..5984
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6012..6252,1..8)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6012..6032
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6183..6202
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gaaacaccgt gctcgcttcg gcagcacata tactagtgat tgatgttttt ttcgacggcc
       61 atcagtcgcc ggtcccaagc ccggataaaa tgggaggggg cgggaaaccg cctaaccatg
      121 ccgactgatg gcaggcccgg atagctcagt cggtagagca gcggccgcgc gattcgagga
      181 aggaggtctg aggaggtcac tgaatcgcgc ggccgcgggt ccagggttca agtccctgtt
      241 cgggcctgcc atcagtcggc gtggactgta gaacactgcc aatgccggtc ccaagcccgg
      301 ataaaagtgg agggtacagt ccacgctcta gagcggactt cggtccgctt tttactagga
      361 cctgcaggca tgcaagcttg acgtcggtta ccgatatcca tatggcggcc gcatcgatct
      421 cgagccgcgg actagtaact tgtttattgc agcttataat ggttacaaat aaagcaatag
      481 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
      541 actcatcaat gtatcttatc atgtcttacg tagataagta gcatggcggg ttaatcatta
      601 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
      661 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
      721 gcgagcgagc gcgcagagag ggacagatct gcagatctgc tggcgtaata gcgaagaggc
      781 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga aattgtaagc
      841 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
      901 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
      961 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     1021 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     1081 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     1141 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     1201 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     1261 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     1321 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1381 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1441 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1501 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1561 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1621 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     1681 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     1741 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     1801 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     1861 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     1921 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     1981 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2041 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2101 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2161 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2221 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2281 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2341 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2401 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2461 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2521 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2581 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2641 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2701 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2761 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2821 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2881 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2941 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3001 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3061 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3121 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3181 accgagcgca gcgagtcagt gagcgagcgc gcagagaggg agtggccaac tccatcacta
     3241 ggggttcctg gaggggtgga gtcgtgacgt gaattacgtc atagggttag ggaggtcctg
     3301 gatcgatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3361 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3421 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3481 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     3541 tgatctctag tcaaggcact atacatcaaa tattccttat taaccccttt acaaattaaa
     3601 aagctaaagg tacacaattt ttgagcatag ttattaatag cagacactct atgcctgtgt
     3661 ggagtaagaa aaaacagtat gttatgatta taactgttat gcctacttat aaaggttaca
     3721 gaatattttt ccataatttt cttgtatagc agtgcagctt tttcctttgt ggtgtaaata
     3781 gcaaagcaag caagagttct attactaaac acagcatgac tcaaaaaact tagcaattct
     3841 gaaggaaagt ccttggggtc ttctaccttt ctcttctttt ttggaggagt agaatgttga
     3901 gagtcagcag tagcctcatc atcactagat ggcatttctt ctgagcaaaa caggttttcc
     3961 tcattaaagg cattccacca ctgctcccat tcatcagttc cataggttgg aatctaaaat
     4021 acacaaacaa ttagaatcag tagtttaaca cattatacac ttaaaaattt tatatttacc
     4081 ttagagcttt aaatctctgt aggtagtttg tccaattatg tcacaccaca gaagtaaggt
     4141 tccttcacaa agatccggac caaagcggcc atcgtgcctc cccactcctg cagttcgggg
     4201 gcatggatgc gcggatagcc gctgctggtt tcctggatgc cgacggattt gcactgccgg
     4261 tagaactccg cgaggtcgtc cagcctcagg cagcagctga accaactcgc gaggggatcg
     4321 agcccggggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
     4381 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
     4441 tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc
     4501 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     4561 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     4621 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
     4681 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     4741 cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga
     4801 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     4861 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     4921 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     4981 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     5041 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     5101 ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     5161 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     5221 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     5281 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatctt gatcccctgc
     5341 gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct
     5401 taccagaggg cgccccagct ggcaattccg gttcgcttgc tgtccataaa accgcccagt
     5461 ctagctatcg ccatgtaagc ccactgcaag ctacctgctt tctctttgcg cttgcgtttt
     5521 cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac
     5581 tggctttcta cgtgttccgc ttcctttagc agcccttgcg ccctgagtgc ttgcggcagc
     5641 gtgaagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     5701 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     5761 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     5821 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     5881 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5941 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagcc ggatccaagg
     6001 tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     6061 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
     6121 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     6181 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     6241 tgtggaaagg ac
//
