LOCUS       sequence-311780-        6268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impacts 4 uracils 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 6268)
  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 6268)
  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(4)_HairpinDistance(10)nts_Tornado-Corn
FEATURES             Location/Qualifiers
     source          1..6268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..264
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     16..36
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     187..206
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    709..830
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   879..1008
                     /label=AAV2 ITR
     rep_origin      complement(1102..1557)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1239..1260)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1451..1470
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1584..1688
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1689..1757,1758..2549)
                     /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(1907..1926)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2720..3308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3209..3228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3577..3711
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4136..4156)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4286..4351
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4771..5565)
                     /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(4882..4901)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5492..5511
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5922..6251)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5936..6071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6236..6256
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aaggtcgggc aggaagaggg cctatttccc atgattcctt catatttgca tatacgatac
       61 aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa
      121 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
      181 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
      241 atcttgtgga aaggacgaaa caccgtgctc gcttcggcag cacatatggt tgcggctttc
      301 tgctgcaatc actagttggc ttttcgacgg ccatcagtcg ccggtcccaa gcccggataa
      361 aatgggaggg ggcgggaaac cgcctaacca tgccgactga tggcaggccc ggatagctca
      421 gtcggtagag cagcggccgc gcgattcgag gaaggaggtc tgaggaggtc actgaatcgc
      481 gcggccgcgg gtccagggtt caagtccctg ttcgggcctg ccatcagtcg gcgtggactg
      541 tagaacactg ccaatgccgg tcccaagccc ggataaaagt ggagggtaca gtccacgctc
      601 tagagcggac ttcggtccgc tttttactag gacctgcagg catgcaagct tgacgtcggt
      661 taccgatatc catatggcgg ccgcatcgat ctcgagccgc ggactagtaa cttgtttatt
      721 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
      781 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctta
      841 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
      901 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
      961 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagag agggacagat
     1021 ctgcagatct gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     1081 cgcagcctga atggcgaatg gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1141 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1201 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1261 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1321 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1381 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1441 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1501 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     1561 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1621 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1681 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1741 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1801 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1861 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     1921 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1981 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2041 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2101 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2161 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2221 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2281 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2341 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2401 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2461 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2521 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2581 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2641 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2701 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2761 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2821 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2881 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2941 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3001 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3061 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3121 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3181 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3241 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3301 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3361 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3421 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagc
     3481 gcgcagagag ggagtggcca actccatcac taggggttcc tggaggggtg gagtcgtgac
     3541 gtgaattacg tcatagggtt agggaggtcc tggatcgatc cagacatgat aagatacatt
     3601 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3661 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3721 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     3781 taaaacctct acaaatgtgg tatggctgat tatgatctct agtcaaggca ctatacatca
     3841 aatattcctt attaacccct ttacaaatta aaaagctaaa ggtacacaat ttttgagcat
     3901 agttattaat agcagacact ctatgcctgt gtggagtaag aaaaaacagt atgttatgat
     3961 tataactgtt atgcctactt ataaaggtta cagaatattt ttccataatt ttcttgtata
     4021 gcagtgcagc tttttccttt gtggtgtaaa tagcaaagca agcaagagtt ctattactaa
     4081 acacagcatg actcaaaaaa cttagcaatt ctgaaggaaa gtccttgggg tcttctacct
     4141 ttctcttctt ttttggagga gtagaatgtt gagagtcagc agtagcctca tcatcactag
     4201 atggcatttc ttctgagcaa aacaggtttt cctcattaaa ggcattccac cactgctccc
     4261 attcatcagt tccataggtt ggaatctaaa atacacaaac aattagaatc agtagtttaa
     4321 cacattatac acttaaaaat tttatattta ccttagagct ttaaatctct gtaggtagtt
     4381 tgtccaatta tgtcacacca cagaagtaag gttccttcac aaagatccgg accaaagcgg
     4441 ccatcgtgcc tccccactcc tgcagttcgg gggcatggat gcgcggatag ccgctgctgg
     4501 tttcctggat gccgacggat ttgcactgcc ggtagaactc cgcgaggtcg tccagcctca
     4561 ggcagcagct gaaccaactc gcgaggggat cgagcccggg gtgggcgaag aactccagca
     4621 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     4681 acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt
     4741 ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa
     4801 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     4861 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     4921 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     4981 attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc
     5041 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     5101 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     5161 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     5221 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     5281 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     5341 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc
     5401 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     5461 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     5521 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
     5581 tcctgtctct tgatcagatc ttgatcccct gcgccatcag atccttggcg gcaagaaagc
     5641 catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag ctggcaattc
     5701 cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa gcccactgca
     5761 agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac
     5821 attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtgttcc gcttccttta
     5881 gcagcccttg cgccctgagt gcttgcggca gcgtgaagct ttttgcaaaa gcctaggcct
     5941 ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct
     6001 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
     6061 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg
     6121 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa
     6181 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
     6241 aactgacaca cattccacag ccggatcc
//
