LOCUS       sequence-330471-        6067 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 4 uracil in a poly-uracil tract on human 
            polymerase III transcription from a GLN tRNA promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6067)
  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 6067)
  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: GLN, Linear-3, PolyU(4)
FEATURES             Location/Qualifiers
     source          1..6067
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..795)
                     /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(112..131)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     722..741
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1152..1481)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1166..1301
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1466..1486
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    2006..2127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2176..2305
                     /label=AAV2 ITR
     rep_origin      complement(2399..2854)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2536..2557)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2748..2767
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2881..2985
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2986..3054,3055..3846)
                     /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(3204..3223)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4017..4605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4506..4525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4874..5008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5433..5453)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5583..5648
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
ORIGIN
        1 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
       61 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
      121 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
      181 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
      241 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
      301 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
      361 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
      421 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
      481 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
      541 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
      601 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
      661 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
      721 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
      781 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct
      841 gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac
      901 cttaccagag ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca
      961 gtctagctat cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt
     1021 ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg
     1081 actggctttc tacgtgttcc gcttccttta gcagcccttg cgccctgagt gcttgcggca
     1141 gcgtgaagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
     1201 agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg
     1261 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     1321 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     1381 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     1441 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ccggatccaa
     1501 ggtcgggcag gaagtctctc gtcggttcca tggtgtaatg gttagcactc tggactctga
     1561 atccagcgat ccgagttcaa atctcggtgg aacctatata ctagcagcca actccaattt
     1621 tcgacggcca tcagtcgccg gtcccaagcc cggataaaat gggagggggc gggaaaccgc
     1681 ctaaccatgc cgactgatgg caggcccgga tagctcagtc ggtagagcag cggccgcgcg
     1741 attcgaggaa ggaggtctga ggaggtcact gaatcgcgcg gccgcgggtc cagggttcaa
     1801 gtccctgttc gggcctgcca tcagtcggcg tggactgtag aacactgcca atgccggtcc
     1861 caagcccgga taaaagtgga gggtacagtc cacgctctag agcggacttc ggtccgcttt
     1921 ttactaggac ctgcaggcat gcaagcttga cgtcggttac cgatatccat atggcggccg
     1981 catcgatctc gagccgcgga ctagtaactt gtttattgca gcttataatg gttacaaata
     2041 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2101 tttgtccaaa ctcatcaatg tatcttatca tgtcttacgt agataagtag catggcgggt
     2161 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     2221 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     2281 cctcagtgag cgagcgagcg cgcagagagg gacagatctg cagatctgct ggcgtaatag
     2341 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggaa
     2401 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2461 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     2521 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2581 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     2641 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2701 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2761 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2821 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     2881 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2941 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3001 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3061 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3121 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3181 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3241 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3301 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3361 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3421 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3481 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3541 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3601 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3661 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3721 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3781 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3841 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3901 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3961 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4021 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4081 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4141 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4201 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4261 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4321 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4381 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4441 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4501 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4561 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4621 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4681 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4741 agccgaacga ccgagcgcag cgagtcagtg agcgagcgcg cagagaggga gtggccaact
     4801 ccatcactag gggttcctgg aggggtggag tcgtgacgtg aattacgtca tagggttagg
     4861 gaggtcctgg atcgatccag acatgataag atacattgat gagtttggac aaaccacaac
     4921 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4981 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     5041 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     5101 ggctgattat gatctctagt caaggcacta tacatcaaat attccttatt aaccccttta
     5161 caaattaaaa agctaaaggt acacaatttt tgagcatagt tattaatagc agacactcta
     5221 tgcctgtgtg gagtaagaaa aaacagtatg ttatgattat aactgttatg cctacttata
     5281 aaggttacag aatatttttc cataattttc ttgtatagca gtgcagcttt ttcctttgtg
     5341 gtgtaaatag caaagcaagc aagagttcta ttactaaaca cagcatgact caaaaaactt
     5401 agcaattctg aaggaaagtc cttggggtct tctacctttc tcttcttttt tggaggagta
     5461 gaatgttgag agtcagcagt agcctcatca tcactagatg gcatttcttc tgagcaaaac
     5521 aggttttcct cattaaaggc attccaccac tgctcccatt catcagttcc ataggttgga
     5581 atctaaaata cacaaacaat tagaatcagt agtttaacac attatacact taaaaatttt
     5641 atatttacct tagagcttta aatctctgta ggtagtttgt ccaattatgt cacaccacag
     5701 aagtaaggtt ccttcacaaa gatccggacc aaagcggcca tcgtgcctcc ccactcctgc
     5761 agttcggggg catggatgcg cggatagccg ctgctggttt cctggatgcc gacggatttg
     5821 cactgccggt agaactccgc gaggtcgtcc agcctcaggc agcagctgaa ccaactcgcg
     5881 aggggatcga gcccggggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc
     5941 atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg
     6001 gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga
     6061 gtcccgc
//
