LOCUS       sequence-313478-        6243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 2 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 6243)
  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 6243)
  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(2)
FEATURES             Location/Qualifiers
     source          1..6243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   35..164
                     /label=AAV2 ITR
     rep_origin      complement(258..713)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(395..416)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     607..626
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        740..844
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(845..913,914..1705)
                     /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(1063..1082)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1876..2464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2365..2384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2733..2867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3292..3312)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          3442..3507
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(3927..4721)
                     /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(4038..4057)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4648..4667
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5078..5407)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5092..5227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5392..5412
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5440..5688
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5440..5460
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5611..5630
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    6108..6229
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc
       61 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
      121 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagagaggg acagatctgc
      181 agatctgctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
      241 gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
      301 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
      361 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
      421 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
      481 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
      541 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
      601 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
      661 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
      721 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      781 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      841 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      901 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      961 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1021 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1081 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1141 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1201 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1261 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1321 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1381 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1441 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1501 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1561 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1621 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1681 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1741 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1801 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1861 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1921 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1981 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2041 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2101 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2161 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2221 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2281 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2341 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2401 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2461 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2521 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2581 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgagcgcgc
     2641 agagagggag tggccaactc catcactagg ggttcctgga ggggtggagt cgtgacgtga
     2701 attacgtcat agggttaggg aggtcctgga tcgatccaga catgataaga tacattgatg
     2761 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2821 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2881 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     2941 acctctacaa atgtggtatg gctgattatg atctctagtc aaggcactat acatcaaata
     3001 ttccttatta acccctttac aaattaaaaa gctaaaggta cacaattttt gagcatagtt
     3061 attaatagca gacactctat gcctgtgtgg agtaagaaaa aacagtatgt tatgattata
     3121 actgttatgc ctacttataa aggttacaga atatttttcc ataattttct tgtatagcag
     3181 tgcagctttt tcctttgtgg tgtaaatagc aaagcaagca agagttctat tactaaacac
     3241 agcatgactc aaaaaactta gcaattctga aggaaagtcc ttggggtctt ctacctttct
     3301 cttctttttt ggaggagtag aatgttgaga gtcagcagta gcctcatcat cactagatgg
     3361 catttcttct gagcaaaaca ggttttcctc attaaaggca ttccaccact gctcccattc
     3421 atcagttcca taggttggaa tctaaaatac acaaacaatt agaatcagta gtttaacaca
     3481 ttatacactt aaaaatttta tatttacctt agagctttaa atctctgtag gtagtttgtc
     3541 caattatgtc acaccacaga agtaaggttc cttcacaaag atccggacca aagcggccat
     3601 cgtgcctccc cactcctgca gttcgggggc atggatgcgc ggatagccgc tgctggtttc
     3661 ctggatgccg acggatttgc actgccggta gaactccgcg aggtcgtcca gcctcaggca
     3721 gcagctgaac caactcgcga ggggatcgag cccggggtgg gcgaagaact ccagcatgag
     3781 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     3841 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     3901 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     3961 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     4021 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
     4081 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     4141 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
     4201 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     4261 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     4321 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     4381 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     4441 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     4501 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     4561 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     4621 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     4681 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     4741 gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc
     4801 cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt
     4861 tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct
     4921 acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     4981 atccggggtc agcaccgttt ctgcggactg gctttctacg tgttccgctt cctttagcag
     5041 cccttgcgcc ctgagtgctt gcggcagcgt gaagcttttt gcaaaagcct aggcctccaa
     5101 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     5161 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     5221 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     5281 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
     5341 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
     5401 gacacacatt ccacagccgg atccaaggtc gggcaggaag agggcctatt tcccatgatt
     5461 ccttcatatt tgcatatacg atacaaggct gttagagaga taattagaat taatttgact
     5521 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
     5581 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
     5641 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgt gctcgcttcg
     5701 gcagcacata tactagttgg cttcgacggc catcagtcgc cggtcccaag cccggataaa
     5761 atgggagggg gcgggaaacc gcctaaccat gccgactgat ggcaggcccg gatagctcag
     5821 tcggtagagc agcggccgcg cgattcgagg aaggaggtct gaggaggtca ctgaatcgcg
     5881 cggccgcggg tccagggttc aagtccctgt tcgggcctgc catcagtcgg cgtggactgt
     5941 agaacactgc caatgccggt cccaagcccg gataaaagtg gagggtacag tccacgctct
     6001 agagcggact tcggtccgct ttttactagg acctgcaggc atgcaagctt gacgtcggtt
     6061 accgatatcc atatggcggc cgcatcgatc tcgagccgcg gactagtaac ttgtttattg
     6121 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6181 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtcttac
     6241 gta
//
