LOCUS       sequence-311770-        6247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 6 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 6247)
  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 6247)
  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(6)_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     249..269
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        297..545
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     297..317
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     468..487
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    969..1090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   1139..1268
                     /label=AAV2 ITR
     rep_origin      complement(1362..1817)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1499..1520)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1711..1730
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1844..1948
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1949..2017,2018..2809)
                     /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(2167..2186)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2980..3568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3469..3488
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3837..3971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4396..4416)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4546..4611
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(5031..5825)
                     /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
                     GLAPAELLARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5142..5161)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5752..5771
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6182..6247,1..264))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6196..6247,1..84)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
       61 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
      121 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
      181 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      241 ctttccacac cctaactgac acacattcca cagccggatc caaggtcggg caggaagagg
      301 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
      361 ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
      421 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
      481 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
      541 acaccgtgct cgcttcggca gcacatatac tagttggctt ttttcgacgg ccatcagtcg
      601 ccggtcccaa gcccggataa aatgggaggg ggcgggaaac cgcctaacca tgccgactga
      661 tggcaggccc ggatagctca gtcggtagag cagcggccgc gcgattcgag gaaggaggtc
      721 tgaggaggtc actgaatcgc gcggccgcgg gtccagggtt caagtccctg ttcgggcctg
      781 ccatcagtcg gcgtggactg tagaacactg ccaatgccgg tcccaagccc ggataaaagt
      841 ggagggtaca gtccacgctc tagagcggac ttcggtccgc tttttactag gacctgcagg
      901 catgcaagct tgacgtcggt taccgatatc catatggcgg ccgcatcgat ctcgagccgc
      961 ggactagtaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     1021 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     1081 atgtatctta tcatgtctta cgtagataag tagcatggcg ggttaatcat taactacaag
     1141 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     1201 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     1261 gcgcgcagag agggacagat ctgcagatct gctggcgtaa tagcgaagag gcccgcaccg
     1321 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaaattgtaa gcgttaatat
     1381 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1441 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     1501 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1561 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     1621 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     1681 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     1741 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     1801 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1861 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1921 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     1981 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2041 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2101 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2161 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     2221 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2281 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2341 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2401 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2461 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2521 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2581 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2641 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2701 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2761 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2821 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     2881 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     2941 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3001 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3061 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3121 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3181 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3241 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3301 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3361 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3421 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3481 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3541 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3601 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3661 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3721 cagcgagtca gtgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     3781 tggaggggtg gagtcgtgac gtgaattacg tcatagggtt agggaggtcc tggatcgatc
     3841 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     3901 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     3961 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     4021 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatctct
     4081 agtcaaggca ctatacatca aatattcctt attaacccct ttacaaatta aaaagctaaa
     4141 ggtacacaat ttttgagcat agttattaat agcagacact ctatgcctgt gtggagtaag
     4201 aaaaaacagt atgttatgat tataactgtt atgcctactt ataaaggtta cagaatattt
     4261 ttccataatt ttcttgtata gcagtgcagc tttttccttt gtggtgtaaa tagcaaagca
     4321 agcaagagtt ctattactaa acacagcatg actcaaaaaa cttagcaatt ctgaaggaaa
     4381 gtccttgggg tcttctacct ttctcttctt ttttggagga gtagaatgtt gagagtcagc
     4441 agtagcctca tcatcactag atggcatttc ttctgagcaa aacaggtttt cctcattaaa
     4501 ggcattccac cactgctccc attcatcagt tccataggtt ggaatctaaa atacacaaac
     4561 aattagaatc agtagtttaa cacattatac acttaaaaat tttatattta ccttagagct
     4621 ttaaatctct gtaggtagtt tgtccaatta tgtcacacca cagaagtaag gttccttcac
     4681 aaagatccgg accaaagcgg ccatcgtgcc tccccactcc tgcagttcgg gggcatggat
     4741 gcgcggatag ccgctgctgg tttcctggat gccgacggat ttgcactgcc ggtagaactc
     4801 cgcgaggtcg tccagcctca ggcagcagct gaaccaactc gcgaggggat cgagcccggg
     4861 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
     4921 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga
     4981 tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac
     5041 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     5101 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     5161 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     5221 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
     5281 tcgccgtcgg gcatgcgcgc cttgagcctg gctaacagtt cggctggcgc gagcccctga
     5341 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     5401 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     5461 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     5521 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     5581 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     5641 tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     5701 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     5761 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     5821 atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag
     5881 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     5941 ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat
     6001 cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc
     6061 cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc
     6121 tacgtgttcc gcttccttta gcagcccttg cgccctgagt gcttgcggca gcgtgaagct
     6181 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     6241 ccgaggc
//
