LOCUS       sequence-311752-        6249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 8 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 6249)
  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 6249)
  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(8)_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    434..555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   604..733
                     /label=AAV2 ITR
     rep_origin      complement(827..1282)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(964..985)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1176..1195
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1309..1413
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1414..1482,1483..2274)
                     /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(1632..1651)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2445..3033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2934..2953
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3302..3436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3861..3881)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4011..4076
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4496..5290)
                     /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(4607..4626)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5217..5236
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5647..5976)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5661..5796
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5961..5981
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6009..6249,1..8)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6009..6029
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6180..6199
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gaaacaccgt gctcgcttcg gcagcacata tactagttgg cttttttttc gacggccatc
       61 agtcgccggt cccaagcccg gataaaatgg gagggggcgg gaaaccgcct aaccatgccg
      121 actgatggca ggcccggata gctcagtcgg tagagcagcg gccgcgcgat tcgaggaagg
      181 aggtctgagg aggtcactga atcgcgcggc cgcgggtcca gggttcaagt ccctgttcgg
      241 gcctgccatc agtcggcgtg gactgtagaa cactgccaat gccggtccca agcccggata
      301 aaagtggagg gtacagtcca cgctctagag cggacttcgg tccgcttttt actaggacct
      361 gcaggcatgc aagcttgacg tcggttaccg atatccatat ggcggccgca tcgatctcga
      421 gccgcggact agtaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
      481 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
      541 catcaatgta tcttatcatg tcttacgtag ataagtagca tggcgggtta atcattaact
      601 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
      661 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
      721 agcgagcgcg cagagaggga cagatctgca gatctgctgg cgtaatagcg aagaggcccg
      781 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggaaat tgtaagcgtt
      841 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
      901 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
      961 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     1021 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     1081 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     1141 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     1201 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     1261 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     1321 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1381 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1441 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1501 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1561 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1621 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     1681 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1741 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1801 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     1861 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     1921 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     1981 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2041 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2101 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2161 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2221 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2281 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2341 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2401 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2461 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2521 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2581 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2641 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2701 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2761 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2821 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2881 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2941 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3001 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3061 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3121 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3181 gagcgcagcg agtcagtgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
     3241 gttcctggag gggtggagtc gtgacgtgaa ttacgtcata gggttaggga ggtcctggat
     3301 cgatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3361 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3421 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3481 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     3541 tctctagtca aggcactata catcaaatat tccttattaa cccctttaca aattaaaaag
     3601 ctaaaggtac acaatttttg agcatagtta ttaatagcag acactctatg cctgtgtgga
     3661 gtaagaaaaa acagtatgtt atgattataa ctgttatgcc tacttataaa ggttacagaa
     3721 tatttttcca taattttctt gtatagcagt gcagcttttt cctttgtggt gtaaatagca
     3781 aagcaagcaa gagttctatt actaaacaca gcatgactca aaaaacttag caattctgaa
     3841 ggaaagtcct tggggtcttc tacctttctc ttcttttttg gaggagtaga atgttgagag
     3901 tcagcagtag cctcatcatc actagatggc atttcttctg agcaaaacag gttttcctca
     3961 ttaaaggcat tccaccactg ctcccattca tcagttccat aggttggaat ctaaaataca
     4021 caaacaatta gaatcagtag tttaacacat tatacactta aaaattttat atttacctta
     4081 gagctttaaa tctctgtagg tagtttgtcc aattatgtca caccacagaa gtaaggttcc
     4141 ttcacaaaga tccggaccaa agcggccatc gtgcctcccc actcctgcag ttcgggggca
     4201 tggatgcgcg gatagccgct gctggtttcc tggatgccga cggatttgca ctgccggtag
     4261 aactccgcga ggtcgtccag cctcaggcag cagctgaacc aactcgcgag gggatcgagc
     4321 ccggggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
     4381 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
     4441 cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga
     4501 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     4561 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     4621 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     4681 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     4741 gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     4801 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     4861 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     4921 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     4981 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     5041 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     5101 cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     5161 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     5221 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     5281 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatcttgat cccctgcgcc
     5341 atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac
     5401 cagagggcgc cccagctggc aattccggtt cgcttgctgt ccataaaacc gcccagtcta
     5461 gctatcgcca tgtaagccca ctgcaagcta cctgctttct ctttgcgctt gcgttttccc
     5521 ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg
     5581 ctttctacgt gttccgcttc ctttagcagc ccttgcgccc tgagtgcttg cggcagcgtg
     5641 aagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     5701 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     5761 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     5821 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     5881 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5941 ggagcctggg gactttccac accctaactg acacacattc cacagccgga tccaaggtcg
     6001 ggcaggaaga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
     6061 ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt
     6121 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
     6181 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
     6241 ggaaaggac
//
