LOCUS       sequence-330474-        6063 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 GLNYtRNA promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6063)
  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 6063)
  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: GLY, Linear-3, PolyU(1)
FEATURES             Location/Qualifiers
     source          1..6063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   87..216
                     /label=AAV2 ITR
     polyA_signal    265..386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(902..922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        907..1236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1087..1222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1593..2387
                     /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(1647..1666)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2257..2276
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     intron          2807..2872
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3002..3022
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3447..3581
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3850..4438)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3930..3949)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4609..5400,5401..5469))
                     /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     5232..5251
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5470..5574)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5601..6056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5688..5707)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5898..5919
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
       61 cagcagatct gcagatctgt ccctctctgc gcgctcgctc gctcactgag gccgcccggg
      121 caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
      181 gagagggagt ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat
      241 gctacttatc tacgtaagac atgataagat acattgatga gtttggacaa accacaacta
      301 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
      361 ccattataag ctgcaataaa caagttacta gtccgcggct cgagatcgat gcggccgcca
      421 tatggatatc ggtaaccgac gtcaagcttg catgcctgca ggtcctagta aaaagcggac
      481 cgaagtccgc tctagagcgt ggactgtacc ctccactttt atccgggctt gggaccggca
      541 ttggcagtgt tctacagtcc acgccgactg atggcaggcc cgaacaggga cttgaaccct
      601 ggacccgcgg ccgcgcgatt cagtgacctc ctcagacctc cttcctcgaa tcgcgcggcc
      661 gctgctctac cgactgagct atccgggcct gccatcagtc ggcatggtta ggcggtttcc
      721 cgccccctcc cattttatcc gggcttggga ccggcgactg atggccgtcg attggagttg
      781 gctgctagta tattgcattg gccgggaatc gaacccgggc ctcccgcgtg gcaggcgaga
      841 attctaccac tgaaccacca atgcgacgag agacttcctg cccgaccttg gatccggctg
      901 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      961 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     1021 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     1081 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     1141 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     1201 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttcacgctgc cgcaagcact
     1261 cagggcgcaa gggctgctaa aggaagcgga acacgtagaa agccagtccg cagaaacggt
     1321 gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa
     1381 agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat
     1441 ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct
     1501 gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat
     1561 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     1621 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     1681 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     1741 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     1801 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     1861 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     1921 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     1981 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     2041 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     2101 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     2161 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     2221 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     2281 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     2341 attcgcagca catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     2401 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     2461 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     2521 ccagcgcggg gatctcatgc tggagttctt cgcccacccc gggctcgatc ccctcgcgag
     2581 ttggttcagc tgctgcctga ggctggacga cctcgcggag ttctaccggc agtgcaaatc
     2641 cgtcggcatc caggaaacca gcagcggcta tccgcgcatc catgcccccg aactgcagga
     2701 gtggggaggc acgatggccg ctttggtccg gatctttgtg aaggaacctt acttctgtgg
     2761 tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa atataaaatt
     2821 tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt agattccaac
     2881 ctatggaact gatgaatggg agcagtggtg gaatgccttt aatgaggaaa acctgttttg
     2941 ctcagaagaa atgccatcta gtgatgatga ggctactgct gactctcaac attctactcc
     3001 tccaaaaaag aagagaaagg tagaagaccc caaggacttt ccttcagaat tgctaagttt
     3061 tttgagtcat gctgtgttta gtaatagaac tcttgcttgc tttgctattt acaccacaaa
     3121 ggaaaaagct gcactgctat acaagaaaat tatggaaaaa tattctgtaa cctttataag
     3181 taggcataac agttataatc ataacatact gttttttctt actccacaca ggcatagagt
     3241 gtctgctatt aataactatg ctcaaaaatt gtgtaccttt agctttttaa tttgtaaagg
     3301 ggttaataag gaatatttga tgtatagtgc cttgactaga gatcataatc agccatacca
     3361 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3421 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3481 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3541 gtttgtccaa actcatcaat gtatcttatc atgtctggat cgatccagga cctccctaac
     3601 cctatgacgt aattcacgtc acgactccac ccctccagga acccctagtg atggagttgg
     3661 ccactccctc tctgcgcgct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3721 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3781 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3841 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3901 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3961 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4021 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4081 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4141 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4201 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4261 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4321 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4381 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4441 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4501 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4561 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4621 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4681 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4741 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4801 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4861 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4921 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4981 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5041 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5101 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5161 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5221 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5281 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5341 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5401 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5461 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5521 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5581 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5641 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5701 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5761 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5821 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5881 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5941 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6001 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat
     6061 tcg
//
