LOCUS       sequence-311777-        6269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impacts 1 uracil 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 6269)
  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 6269)
  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(1)_HairpinDistance(10)nts_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    695..816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   865..994
                     /label=AAV2 ITR
     rep_origin      complement(1088..1543)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1225..1246)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1437..1456
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1570..1674
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1675..1743,1744..2535)
                     /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(1893..1912)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2706..3294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3195..3214
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3563..3697
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4122..4142)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4272..4337
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4757..5551)
                     /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(4868..4887)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5478..5497
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5908..6237)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5922..6057
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6222..6242
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg tgctcgcttc ggcagcacat atactaggtt gcggctttct gctgcaatcg
      301 tgattgatgt cgacggccat cagtcgccgg tcccaagccc ggataaaatg ggagggggcg
      361 ggaaaccgcc taaccatgcc gactgatggc aggcccggat agctcagtcg gtagagcagc
      421 ggccgcgcga ttcgaggaag gaggtctgag gaggtcactg aatcgcgcgg ccgcgggtcc
      481 agggttcaag tccctgttcg ggcctgccat cagtcggcgt ggactgtaga acactgccaa
      541 tgccggtccc aagcccggat aaaagtggag ggtacagtcc acgctctaga gcggacttcg
      601 gtccgctttt tactaggacc tgcaggcatg caagcttgac gtcggttacc gatatccata
      661 tggcggccgc atcgatctcg agccgcggac tagtaacttg tttattgcag cttataatgg
      721 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
      781 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtcttacgta gataagtagc
      841 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
      901 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
      961 cccgggcggc ctcagtgagc gagcgagcgc gcagagaggg acagatctgc agatctgctg
     1021 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     1081 cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     1141 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     1201 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     1261 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     1321 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     1381 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     1441 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     1501 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     1561 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1621 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1681 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1741 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1801 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1861 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1921 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1981 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2041 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2101 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2161 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2221 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2281 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2341 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2401 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2461 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2521 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2581 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2641 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2701 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2761 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2821 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2881 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2941 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3001 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3061 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3121 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3181 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3241 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3301 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3361 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3421 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgagcgcgc agagagggag
     3481 tggccaactc catcactagg ggttcctgga ggggtggagt cgtgacgtga attacgtcat
     3541 agggttaggg aggtcctgga tcgatccaga catgataaga tacattgatg agtttggaca
     3601 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3661 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3721 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     3781 atgtggtatg gctgattatg atctctagtc aaggcactat acatcaaata ttccttatta
     3841 acccctttac aaattaaaaa gctaaaggta cacaattttt gagcatagtt attaatagca
     3901 gacactctat gcctgtgtgg agtaagaaaa aacagtatgt tatgattata actgttatgc
     3961 ctacttataa aggttacaga atatttttcc ataattttct tgtatagcag tgcagctttt
     4021 tcctttgtgg tgtaaatagc aaagcaagca agagttctat tactaaacac agcatgactc
     4081 aaaaaactta gcaattctga aggaaagtcc ttggggtctt ctacctttct cttctttttt
     4141 ggaggagtag aatgttgaga gtcagcagta gcctcatcat cactagatgg catttcttct
     4201 gagcaaaaca ggttttcctc attaaaggca ttccaccact gctcccattc atcagttcca
     4261 taggttggaa tctaaaatac acaaacaatt agaatcagta gtttaacaca ttatacactt
     4321 aaaaatttta tatttacctt agagctttaa atctctgtag gtagtttgtc caattatgtc
     4381 acaccacaga agtaaggttc cttcacaaag atccggacca aagcggccat cgtgcctccc
     4441 cactcctgca gttcgggggc atggatgcgc ggatagccgc tgctggtttc ctggatgccg
     4501 acggatttgc actgccggta gaactccgcg aggtcgtcca gcctcaggca gcagctgaac
     4561 caactcgcga ggggatcgag cccggggtgg gcgaagaact ccagcatgag atccccgcgc
     4621 tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag
     4681 gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg
     4741 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     4801 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     4861 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     4921 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     4981 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga
     5041 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     5101 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     5161 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     5221 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     5281 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     5341 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
     5401 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     5461 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     5521 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     5581 cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt
     5641 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg
     5701 tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct acctgctttc
     5761 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc
     5821 agcaccgttt ctgcggactg gctttctacg tgttccgctt cctttagcag cccttgcgcc
     5881 ctgagtgctt gcggcagcgt gaagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     5941 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     6001 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     6061 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     6121 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     6181 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     6241 ccacagccgg atccaaggtc gggcaggaa
//
