LOCUS       sequence-329692-        6064 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 GLN tRNA promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6064)
  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 6064)
  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(1)
FEATURES             Location/Qualifiers
     source          1..6064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(195..650)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(332..353)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     544..563
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        677..781
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(782..850,851..1642)
                     /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(1000..1019)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1813..2401
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2302..2321
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2670..2804
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3229..3249)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          3379..3444
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(3864..4658)
                     /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
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQHIAFYRLLDEFF"
     primer_bind     complement(3975..3994)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4585..4604
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5015..5344)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5029..5164
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5329..5349
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    5866..5987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   join(6036..6064,1..101)
                     /label=AAV2 ITR
ORIGIN
        1 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
       61 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gagagggaca gatctgcaga
      121 tctgctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
      181 tgaatggcga atggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
      241 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
      301 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
      361 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
      421 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
      481 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
      541 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
      601 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
      661 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      721 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      781 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
      841 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
      901 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
      961 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1021 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1081 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1141 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     1201 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     1261 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1321 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     1381 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     1441 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     1501 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     1561 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     1621 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     1681 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1741 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1801 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1861 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1921 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     1981 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2041 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2101 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2161 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2221 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2281 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2341 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2401 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2461 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     2521 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg agcgcgcaga
     2581 gagggagtgg ccaactccat cactaggggt tcctggaggg gtggagtcgt gacgtgaatt
     2641 acgtcatagg gttagggagg tcctggatcg atccagacat gataagatac attgatgagt
     2701 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     2761 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     2821 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     2881 tctacaaatg tggtatggct gattatgatc tctagtcaag gcactataca tcaaatattc
     2941 cttattaacc cctttacaaa ttaaaaagct aaaggtacac aatttttgag catagttatt
     3001 aatagcagac actctatgcc tgtgtggagt aagaaaaaac agtatgttat gattataact
     3061 gttatgccta cttataaagg ttacagaata tttttccata attttcttgt atagcagtgc
     3121 agctttttcc tttgtggtgt aaatagcaaa gcaagcaaga gttctattac taaacacagc
     3181 atgactcaaa aaacttagca attctgaagg aaagtccttg gggtcttcta cctttctctt
     3241 cttttttgga ggagtagaat gttgagagtc agcagtagcc tcatcatcac tagatggcat
     3301 ttcttctgag caaaacaggt tttcctcatt aaaggcattc caccactgct cccattcatc
     3361 agttccatag gttggaatct aaaatacaca aacaattaga atcagtagtt taacacatta
     3421 tacacttaaa aattttatat ttaccttaga gctttaaatc tctgtaggta gtttgtccaa
     3481 ttatgtcaca ccacagaagt aaggttcctt cacaaagatc cggaccaaag cggccatcgt
     3541 gcctccccac tcctgcagtt cgggggcatg gatgcgcgga tagccgctgc tggtttcctg
     3601 gatgccgacg gatttgcact gccggtagaa ctccgcgagg tcgtccagcc tcaggcagca
     3661 gctgaaccaa ctcgcgaggg gatcgagccc ggggtgggcg aagaactcca gcatgagatc
     3721 cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc
     3781 atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt
     3841 catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg
     3901 tgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg
     3961 ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca
     4021 cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc
     4081 aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc
     4141 ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg
     4201 acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg
     4261 aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat
     4321 actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat
     4381 agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc
     4441 gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac
     4501 aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca
     4561 tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg
     4621 gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc
     4681 tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag
     4741 tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa ttccggttcg
     4801 cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc
     4861 tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
     4921 cggggtcagc accgtttctg cggactggct ttctacgtgt tccgcttcct ttagcagccc
     4981 ttgcgccctg agtgcttgcg gcagcgtgaa gctttttgca aaagcctagg cctccaaaaa
     5041 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     5101 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     5161 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     5221 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     5281 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     5341 acacattcca cagccggatc caaggtcggg caggaagtct ctcgtcggtt ccatggtgta
     5401 atggttagca ctctggactc tgaatccagc gatccgagtt caaatctcgg tggaacctat
     5461 atactagcag ccaactccaa tcgacggcca tcagtcgccg gtcccaagcc cggataaaat
     5521 gggagggggc gggaaaccgc ctaaccatgc cgactgatgg caggcccgga tagctcagtc
     5581 ggtagagcag cggccgcgcg attcgaggaa ggaggtctga ggaggtcact gaatcgcgcg
     5641 gccgcgggtc cagggttcaa gtccctgttc gggcctgcca tcagtcggcg tggactgtag
     5701 aacactgcca atgccggtcc caagcccgga taaaagtgga gggtacagtc cacgctctag
     5761 agcggacttc ggtccgcttt ttactaggac ctgcaggcat gcaagcttga cgtcggttac
     5821 cgatatccat atggcggccg catcgatctc gagccgcgga ctagtaactt gtttattgca
     5881 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5941 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtcttacgt
     6001 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     6061 cact
//
