LOCUS       sequence-308326-        6452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impacts 1 uracil in a poly-uracil tract in 
            conjunctiuon with an immediately upstream hairpin 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 6452)
  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 6452)
  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(0)nts_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     605..624
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1035..1364)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1049..1184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1349..1369
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1397..1645
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1397..1417
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1568..1587
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    2091..2212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2261..2390
                     /label=AAV2 ITR
     rep_origin      complement(2521..2949)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2631..2652)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2843..2862
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2976..3080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3081..3149,3150..3941)
                     /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(3299..3318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4112..4700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4601..4620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5143..5277
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5702..5722)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5852..5917
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     primer_bind     complement(join(6449..6452,1..16))
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
       61 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
      121 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
      181 ggcgcgagcc cctgatgctc ttgtccagat catcctgatc gacaagaccg gcttccatcc
      241 gagtacgtgc tcgctcgatg cgatgttcgc ttggtggtcg aatgggcagg tagccggatc
      301 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
      361 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
      421 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
      481 ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
      541 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
      601 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
      661 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc
      721 cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc
      781 aaccttacca gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc
      841 ccagtctagc tatcggcatg taagcccact gcaagctacc tgctttctct ttgcgcttgc
      901 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
      961 cggactggct ttctacgtgt tccgcttcct ttagcagccc ttgcgccctg agtgcttgcg
     1021 gcagcgtgaa gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     1081 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     1141 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     1201 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     1261 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     1321 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagccggatc
     1381 caaggtcggg caggaagagg gcctatttcc catgattcct tcatatttgc atatacgata
     1441 caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca
     1501 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     1561 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     1621 tatcttgtgg aaaggacgaa acaccgtgct cgcttcggca gcacatatac tagtgattga
     1681 tgggttgcgg ctttctgctg caatctcgac ggccatcagt cgccggtccc aagcccggat
     1741 aaaatgggag ggggcgggaa accgcctaac catgccgact gatggcaggc ccggatagct
     1801 cagtcggtag agcagcggcc gcgcgattcg aggaaggagg tctgaggagg tcactgaatc
     1861 gcgcggccgc gggtccaggg ttcaagtccc tgttcgggcc tgccatcagt cggcgtggac
     1921 tgtagaacac tgccaatgcc ggtcccaagc ccggataaaa gtggagggta cagtccacgc
     1981 tctagagcgg acttcggtcc gctttttact aggacctgca ggcatgcaag cttgacgtcg
     2041 gttaccgata tccatatggc ggccgcatcg atctcgagcc gcggactagt aacttgttta
     2101 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2161 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     2221 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     2281 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     2341 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag agagggagtg
     2401 gccaaagatc tctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2461 cgcagcctga atggctaatg ggaaattgta aacgttaata ttttgttaat attttgttaa
     2521 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2581 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     2641 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2701 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2761 gtaaagcact aaatcggaac cctaaaggga tgccccgatt tagagcttga cggggaaagc
     2821 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     2881 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2941 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3001 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3061 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3121 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3181 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3241 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3301 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3361 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3421 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3481 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3541 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3601 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3661 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3721 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3781 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3841 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3901 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3961 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4021 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4081 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4141 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4201 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4261 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4321 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4381 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4441 cgtgcaacac agccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4501 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4561 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4621 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4681 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4741 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4801 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4861 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     4921 cgattcatta atgcagagat ctttggccac tccctctctg cgcgctcgct cgctcactga
     4981 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     5041 gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgga ggggtggagt
     5101 cgtgacgtga attacgtcat agggttaggg aggtcctgga tcgatccaga catgataaga
     5161 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     5221 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     5281 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     5341 agcaagtaaa acctctacaa atgtggtatg gctgattatg atctctagtc aaggcactat
     5401 acatcaaata ttccttatta acccctttac aaattaaaaa gctaaaggta cacaattttt
     5461 gagcatagtt attaatagca gacactctat gcctgtgtgg agtaagaaaa aacagtatgt
     5521 tatgattata actgttatgc ctacttataa aggttacaga atatttttcc ataattttct
     5581 tgtatagcag tgcagctttt tcctttgtgg tgtaaatagc aaagcaagca agagttctat
     5641 tactaaacac agcatgactc aaaaaactta gcaattctga aggaaagtcc ttggggtctt
     5701 ctacctttct cttctttttt ggaggagtag aatgttgaga gtcagcagta gcctcatcat
     5761 cactagatgg catttcttct gagcaaaaca ggttttcctc attaaaggca ttccaccact
     5821 gctcccattc atcagttcca taggttggaa tctaaaatac acaaacaatt agaatcagta
     5881 gtttaacaca ttatacactt aaaaatttta tatttacctt agagctttaa atctctgtag
     5941 gtagtttgtc caattatgtc acaccacaga agtaaggttc cttcacaaag atccgggacc
     6001 aaagcggcca tcgtgcctcc ccactcctgc agttcggggg catggatgcg cggatagccg
     6061 ctgctggttt cctggatgcc gacggatttg cactgccggt agaactccgc gaggtcgtcc
     6121 agcctcaggc agcagctgaa ccaactcgcg aggggatcga gcccggggtg ggcgaagaac
     6181 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     6241 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
     6301 gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     6361 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     6421 cagcccattc gccgccaagc tcttcagcaa ta
//
