LOCUS       sequence-311729-        6250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 1 uracil 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 6250)
  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 6250)
  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-3_PolyU(1)_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    435..556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   605..734
                     /label=AAV2 ITR
     rep_origin      complement(828..1283)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(965..986)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1177..1196
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1310..1414
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1415..1483,1484..2275)
                     /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(1633..1652)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2446..3034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2935..2954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3303..3437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3862..3882)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4012..4077
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4497..5291)
                     /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(4608..4627)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5218..5237
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5648..5977)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5662..5797
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5962..5982
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6010..6250,1..8)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6010..6030
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6181..6200
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gaaacaccgt gctcgcttcg gcagcacata tactagcagc caactccaat cgacggccat
       61 cagtcgccgg tcccaagccc ggataaaatg ggagggggcg ggaaaccgcc taaccatgcc
      121 gactgatggc aggcccggat agctcagtcg gtagagcagc ggccgcgcga ttcgaggaag
      181 gaggtctgag gaggtcactg aatcgcgcgg ccgcgggtcc agggttcaag tccctgttcg
      241 ggcctgccat cagtcggcgt ggactgtaga acactgccaa tgccggtccc aagcccggat
      301 aaaagtggag ggtacagtcc acgctctaga gcggacttcg gtccgctttt tactaggacc
      361 tgcaggcatg caagcttgac gtcggttacc gatatccata tggcggccgc atcgatctcg
      421 agccgcggac tagtaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
      481 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
      541 tcatcaatgt atcttatcat gtcttacgta gataagtagc atggcgggtt aatcattaac
      601 tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
      661 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
      721 gagcgagcgc gcagagaggg acagatctgc agatctgctg gcgtaatagc gaagaggccc
      781 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt
      841 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
      901 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
      961 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     1021 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     1081 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     1141 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     1201 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     1261 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1321 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1381 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     1441 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1501 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1561 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1621 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     1681 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1741 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1801 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     1861 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     1921 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     1981 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2041 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2101 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     2161 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2221 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     2281 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     2341 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     2401 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2461 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2521 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2581 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2641 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2701 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2761 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2821 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2881 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2941 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3001 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3061 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3121 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     3181 cgagcgcagc gagtcagtga gcgagcgcgc agagagggag tggccaactc catcactagg
     3241 ggttcctgga ggggtggagt cgtgacgtga attacgtcat agggttaggg aggtcctgga
     3301 tcgatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     3361 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3421 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3481 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     3541 atctctagtc aaggcactat acatcaaata ttccttatta acccctttac aaattaaaaa
     3601 gctaaaggta cacaattttt gagcatagtt attaatagca gacactctat gcctgtgtgg
     3661 agtaagaaaa aacagtatgt tatgattata actgttatgc ctacttataa aggttacaga
     3721 atatttttcc ataattttct tgtatagcag tgcagctttt tcctttgtgg tgtaaatagc
     3781 aaagcaagca agagttctat tactaaacac agcatgactc aaaaaactta gcaattctga
     3841 aggaaagtcc ttggggtctt ctacctttct cttctttttt ggaggagtag aatgttgaga
     3901 gtcagcagta gcctcatcat cactagatgg catttcttct gagcaaaaca ggttttcctc
     3961 attaaaggca ttccaccact gctcccattc atcagttcca taggttggaa tctaaaatac
     4021 acaaacaatt agaatcagta gtttaacaca ttatacactt aaaaatttta tatttacctt
     4081 agagctttaa atctctgtag gtagtttgtc caattatgtc acaccacaga agtaaggttc
     4141 cttcacaaag atccggacca aagcggccat cgtgcctccc cactcctgca gttcgggggc
     4201 atggatgcgc ggatagccgc tgctggtttc ctggatgccg acggatttgc actgccggta
     4261 gaactccgcg aggtcgtcca gcctcaggca gcagctgaac caactcgcga ggggatcgag
     4321 cccggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc
     4381 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc
     4441 tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag
     4501 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     4561 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     4621 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     4681 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     4741 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc
     4801 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     4861 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     4921 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     4981 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     5041 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     5101 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     5161 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     5221 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     5281 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatcttga tcccctgcgc
     5341 catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta
     5401 ccagagggcg ccccagctgg caattccggt tcgcttgctg tccataaaac cgcccagtct
     5461 agctatcgcc atgtaagccc actgcaagct acctgctttc tctttgcgct tgcgttttcc
     5521 cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg
     5581 gctttctacg tgttccgctt cctttagcag cccttgcgcc ctgagtgctt gcggcagcgt
     5641 gaagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     5701 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     5761 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     5821 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     5881 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     5941 gggagcctgg ggactttcca caccctaact gacacacatt ccacagccgg atccaaggtc
     6001 gggcaggaag agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     6061 gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg
     6121 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     6181 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
     6241 tggaaaggac
//
