LOCUS       sequence-330470-        6086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 1 uracil in conjunction with a hairpin 
            structure in a poly-uracil tract on human polymerase III 
            transcription from a GLY tRNA promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6086)
  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 6086)
  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), Hairpin Distance = 0nt
FEATURES             Location/Qualifiers
     source          1..6086
                     /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    2025..2146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2195..2324
                     /label=AAV2 ITR
     rep_origin      complement(2418..2873)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2555..2576)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2767..2786
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2900..3004
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3005..3073,3074..3865)
                     /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(3223..3242)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4036..4624
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4525..4544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4893..5027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5452..5472)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5602..5667
                     /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 gtcgcattgg tggttcagtg gtagaattct cgcctgccac
     1561 gcgggaggcc cgggttcgat tcccggccaa tgcaatatac tagcagccaa ctccaaggtt
     1621 gcggctttct gctgcaatct cgacggccat cagtcgccgg tcccaagccc ggataaaatg
     1681 ggagggggcg ggaaaccgcc taaccatgcc gactgatggc aggcccggat agctcagtcg
     1741 gtagagcagc ggccgcgcga ttcgaggaag gaggtctgag gaggtcactg aatcgcgcgg
     1801 ccgcgggtcc agggttcaag tccctgttcg ggcctgccat cagtcggcgt ggactgtaga
     1861 acactgccaa tgccggtccc aagcccggat aaaagtggag ggtacagtcc acgctctaga
     1921 gcggacttcg gtccgctttt tactaggacc tgcaggcatg caagcttgac gtcggttacc
     1981 gatatccata tggcggccgc atcgatctcg agccgcggac tagtaacttg tttattgcag
     2041 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2101 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtcttacgta
     2161 gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc
     2221 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     2281 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagagaggg acagatctgc
     2341 agatctgctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     2401 gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2461 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2521 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2581 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2641 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2701 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2761 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2821 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     2881 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2941 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3001 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3061 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3121 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3181 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3241 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3301 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3361 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3421 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3481 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3541 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3601 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3661 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3721 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3781 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3841 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3901 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3961 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4021 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4081 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4141 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4201 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4261 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4321 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4381 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4441 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4501 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4561 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4621 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4681 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4741 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgagcgcgc
     4801 agagagggag tggccaactc catcactagg ggttcctgga ggggtggagt cgtgacgtga
     4861 attacgtcat agggttaggg aggtcctgga tcgatccaga catgataaga tacattgatg
     4921 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     4981 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     5041 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     5101 acctctacaa atgtggtatg gctgattatg atctctagtc aaggcactat acatcaaata
     5161 ttccttatta acccctttac aaattaaaaa gctaaaggta cacaattttt gagcatagtt
     5221 attaatagca gacactctat gcctgtgtgg agtaagaaaa aacagtatgt tatgattata
     5281 actgttatgc ctacttataa aggttacaga atatttttcc ataattttct tgtatagcag
     5341 tgcagctttt tcctttgtgg tgtaaatagc aaagcaagca agagttctat tactaaacac
     5401 agcatgactc aaaaaactta gcaattctga aggaaagtcc ttggggtctt ctacctttct
     5461 cttctttttt ggaggagtag aatgttgaga gtcagcagta gcctcatcat cactagatgg
     5521 catttcttct gagcaaaaca ggttttcctc attaaaggca ttccaccact gctcccattc
     5581 atcagttcca taggttggaa tctaaaatac acaaacaatt agaatcagta gtttaacaca
     5641 ttatacactt aaaaatttta tatttacctt agagctttaa atctctgtag gtagtttgtc
     5701 caattatgtc acaccacaga agtaaggttc cttcacaaag atccggacca aagcggccat
     5761 cgtgcctccc cactcctgca gttcgggggc atggatgcgc ggatagccgc tgctggtttc
     5821 ctggatgccg acggatttgc actgccggta gaactccgcg aggtcgtcca gcctcaggca
     5881 gcagctgaac caactcgcga ggggatcgag cccggggtgg gcgaagaact ccagcatgag
     5941 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     6001 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     6061 ggtcatttcg aaccccagag tcccgc
//
