LOCUS       sequence-311750-        6248 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 6248)
  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 6248)
  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(7)_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..27
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     repeat_region   258..387
                     /label=AAV2 ITR
     polyA_signal    436..557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(982..1230)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1040..1059)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1210..1230)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1258..1278)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1263..1592
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1443..1578
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1949..2743
                     /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(2003..2022)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2613..2632
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     intron          3163..3228
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3358..3378
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3803..3937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4206..4794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4286..4305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4965..5756,5757..5825))
                     /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     5588..5607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5826..5930)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5957..6248,1..164)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6044..6063)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
       61 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
      121 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg
      181 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagcagatc
      241 tgcagatctg tccctctctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
      301 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
      361 tggccaactc catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat
      421 ctacgtaaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
      481 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
      541 gctgcaataa acaagttact agtccgcggc tcgagatcga tgcggccgcc atatggatat
      601 cggtaaccga cgtcaagctt gcatgcctgc aggtcctagt aaaaagcgga ccgaagtccg
      661 ctctagagcg tggactgtac cctccacttt tatccgggct tgggaccggc attggcagtg
      721 ttctacagtc cacgccgact gatggcaggc ccgaacaggg acttgaaccc tggacccgcg
      781 gccgcgcgat tcagtgacct cctcagacct ccttcctcga atcgcgcggc cgctgctcta
      841 ccgactgagc tatccgggcc tgccatcagt cggcatggtt aggcggtttc ccgccccctc
      901 ccattttatc cgggcttggg accggcgact gatggccgtc gaaaaaaagc caactagtat
      961 atgtgctgcc gaagcgagca cggtgtttcg tcctttccac aagatatata aagccaagaa
     1021 atcgaaatac tttcaagtta cggtaagcat atgatagtcc attttaaaac ataattttaa
     1081 aactgcaaac tacccaagaa attattactt tctacgtcac gtattttgta ctaatatctt
     1141 tgtgtttaca gtcaaattaa ttctaattat ctctctaaca gccttgtatc gtatatgcaa
     1201 atatgaagga atcatgggaa ataggccctc ttcctgcccg accttggatc cggctgtgga
     1261 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     1321 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     1381 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     1441 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     1501 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     1561 gaggcttttt tggaggccta ggcttttgca aaaagcttca cgctgccgca agcactcagg
     1621 gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga aacggtgctg
     1681 accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag
     1741 aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac
     1801 agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
     1861 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga
     1921 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     1981 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     2041 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     2101 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     2161 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     2221 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     2281 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     2341 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     2401 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     2461 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     2521 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     2581 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     2641 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     2701 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     2761 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     2821 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     2881 cgcggggatc tcatgctgga gttcttcgcc caccccgggc tcgatcccct cgcgagttgg
     2941 ttcagctgct gcctgaggct ggacgacctc gcggagttct accggcagtg caaatccgtc
     3001 ggcatccagg aaaccagcag cggctatccg cgcatccatg cccccgaact gcaggagtgg
     3061 ggaggcacga tggccgcttt ggtccggatc tttgtgaagg aaccttactt ctgtggtgtg
     3121 acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta
     3181 agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat
     3241 ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca
     3301 gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca
     3361 aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg
     3421 agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa
     3481 aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt tataagtagg
     3541 cataacagtt ataatcataa catactgttt tttcttactc cacacaggca tagagtgtct
     3601 gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg taaaggggtt
     3661 aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt
     3721 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3781 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3841 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3901 gtccaaactc atcaatgtat cttatcatgt ctggatcgat ccaggacctc cctaacccta
     3961 tgacgtaatt cacgtcacga ctccacccct ccaggaaccc ctagtgatgg agttggccac
     4021 tccctctctg cgcgctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4081 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4141 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4201 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4261 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4321 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4381 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4441 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4501 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4561 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4621 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4681 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4741 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4801 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4861 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4921 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4981 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5041 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5101 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5161 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5221 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5281 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5341 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5401 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5461 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5521 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5581 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5641 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5701 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5761 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5821 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5881 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5941 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     6001 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6061 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6121 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6181 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6241 acgttctt
//
