LOCUS       sequence-330473-        6066 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 GLY tRNA promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6066)
  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 6066)
  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(4)
FEATURES             Location/Qualifiers
     source          1..6066
                     /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
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQHIAFYRLLDEFF"
     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    2005..2126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2175..2304
                     /label=AAV2 ITR
     rep_origin      complement(2398..2853)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2535..2556)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2747..2766
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2880..2984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2985..3053,3054..3845)
                     /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(3203..3222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4016..4604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4505..4524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4873..5007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5432..5452)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5582..5647
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
ORIGIN
        1 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgtgc 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 ctccaatttt
     1621 cgacggccat cagtcgccgg tcccaagccc ggataaaatg ggagggggcg ggaaaccgcc
     1681 taaccatgcc gactgatggc aggcccggat agctcagtcg gtagagcagc ggccgcgcga
     1741 ttcgaggaag gaggtctgag gaggtcactg aatcgcgcgg ccgcgggtcc agggttcaag
     1801 tccctgttcg ggcctgccat cagtcggcgt ggactgtaga acactgccaa tgccggtccc
     1861 aagcccggat aaaagtggag ggtacagtcc acgctctaga gcggacttcg gtccgctttt
     1921 tactaggacc tgcaggcatg caagcttgac gtcggttacc gatatccata tggcggccgc
     1981 atcgatctcg agccgcggac tagtaacttg tttattgcag cttataatgg ttacaaataa
     2041 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     2101 ttgtccaaac tcatcaatgt atcttatcat gtcttacgta gataagtagc atggcgggtt
     2161 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2221 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2281 ctcagtgagc gagcgagcgc gcagagaggg acagatctgc agatctgctg gcgtaatagc
     2341 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa
     2401 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     2461 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     2521 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2581 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     2641 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2701 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     2761 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     2821 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     2881 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2941 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     3001 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3061 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3121 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3181 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3241 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3301 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3361 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3421 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3481 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3541 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3601 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3661 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3721 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3781 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3841 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3901 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3961 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4021 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4081 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4141 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4201 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4261 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4321 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4381 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4441 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4501 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4561 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4621 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4681 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4741 gccgaacgac cgagcgcagc gagtcagtga gcgagcgcgc agagagggag tggccaactc
     4801 catcactagg ggttcctgga ggggtggagt cgtgacgtga attacgtcat agggttaggg
     4861 aggtcctgga tcgatccaga catgataaga tacattgatg agtttggaca aaccacaact
     4921 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4981 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     5041 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     5101 gctgattatg atctctagtc aaggcactat acatcaaata ttccttatta acccctttac
     5161 aaattaaaaa gctaaaggta cacaattttt gagcatagtt attaatagca gacactctat
     5221 gcctgtgtgg agtaagaaaa aacagtatgt tatgattata actgttatgc ctacttataa
     5281 aggttacaga atatttttcc ataattttct tgtatagcag tgcagctttt tcctttgtgg
     5341 tgtaaatagc aaagcaagca agagttctat tactaaacac agcatgactc aaaaaactta
     5401 gcaattctga aggaaagtcc ttggggtctt ctacctttct cttctttttt ggaggagtag
     5461 aatgttgaga gtcagcagta gcctcatcat cactagatgg catttcttct gagcaaaaca
     5521 ggttttcctc attaaaggca ttccaccact gctcccattc atcagttcca taggttggaa
     5581 tctaaaatac acaaacaatt agaatcagta gtttaacaca ttatacactt aaaaatttta
     5641 tatttacctt agagctttaa atctctgtag gtagtttgtc caattatgtc acaccacaga
     5701 agtaaggttc cttcacaaag atccggacca aagcggccat cgtgcctccc cactcctgca
     5761 gttcgggggc atggatgcgc ggatagccgc tgctggtttc ctggatgccg acggatttgc
     5821 actgccggta gaactccgcg aggtcgtcca gcctcaggca gcagctgaac caactcgcga
     5881 ggggatcgag cccggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     5941 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     6001 aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag
     6061 tcccgc
//
