LOCUS       sequence-329691-        6090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of 4 uracil in conjunction with a hairpin 
            structure 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 6090)
  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 6090)
  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), Hairpin Distance = 0nt
FEATURES             Location/Qualifiers
     source          1..6090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   51..180
                     /label=AAV2 ITR
     polyA_signal    229..350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(893..913)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        898..1227
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1078..1213
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1584..2378
                     /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(1638..1657)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2248..2267
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     intron          2798..2863
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2993..3013
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3438..3572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3841..4429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3921..3940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4600..5391,5392..5460))
                     /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     5223..5242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5461..5565)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5592..6047
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5679..5698)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5889..5910
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtgcgggcct cttcgctatt acgccagcag atctgcagat ctgtccctct ctgcgcgctc
       61 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg
      121 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      181 tgtagttaat gattaacccg ccatgctact tatctacgta agacatgata agatacattg
      241 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      301 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt actagtccgc
      361 ggctcgagat cgatgcggcc gccatatgga tatcggtaac cgacgtcaag cttgcatgcc
      421 tgcaggtcct agtaaaaagc ggaccgaagt ccgctctaga gcgtggactg taccctccac
      481 ttttatccgg gcttgggacc ggcattggca gtgttctaca gtccacgccg actgatggca
      541 ggcccgaaca gggacttgaa ccctggaccc gcggccgcgc gattcagtga cctcctcaga
      601 cctccttcct cgaatcgcgc ggccgctgct ctaccgactg agctatccgg gcctgccatc
      661 agtcggcatg gttaggcggt ttcccgcccc ctcccatttt atccgggctt gggaccggcg
      721 actgatggcc gtcgaaaaga ttgcagcaga aagccgcaac cttggagttg gctgctagta
      781 tataggttcc accgagattt gaactcggat cgctggattc agagtccaga gtgctaacca
      841 ttacaccatg gaaccgacga gagacttcct gcccgacctt ggatccggct gtggaatgtg
      901 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
      961 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     1021 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     1081 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     1141 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     1201 ttttttggag gcctaggctt ttgcaaaaag cttcacgctg ccgcaagcac tcagggcgca
     1261 agggctgcta aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg tgctgacccc
     1321 ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc
     1381 aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa
     1441 gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa
     1501 actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga tctgatcaag
     1561 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     1621 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     1681 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     1741 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     1801 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     1861 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     1921 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     1981 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     2041 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     2101 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     2161 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     2221 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     2281 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     2341 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     2401 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     2461 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     2521 ggatctcatg ctggagttct tcgcccaccc cgggctcgat cccctcgcga gttggttcag
     2581 ctgctgcctg aggctggacg acctcgcgga gttctaccgg cagtgcaaat ccgtcggcat
     2641 ccaggaaacc agcagcggct atccgcgcat ccatgccccc gaactgcagg agtggggagg
     2701 cacgatggcc gctttggtcc ggatctttgt gaaggaacct tacttctgtg gtgtgacata
     2761 attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt
     2821 ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa cctatggaac
     2881 tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa aacctgtttt gctcagaaga
     2941 aatgccatct agtgatgatg aggctactgc tgactctcaa cattctactc ctccaaaaaa
     3001 gaagagaaag gtagaagacc ccaaggactt tccttcagaa ttgctaagtt ttttgagtca
     3061 tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc
     3121 tgcactgcta tacaagaaaa ttatggaaaa atattctgta acctttataa gtaggcataa
     3181 cagttataat cataacatac tgttttttct tactccacac aggcatagag tgtctgctat
     3241 taataactat gctcaaaaat tgtgtacctt tagcttttta atttgtaaag gggttaataa
     3301 ggaatatttg atgtatagtg ccttgactag agatcataat cagccatacc acatttgtag
     3361 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3421 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3481 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3541 aactcatcaa tgtatcttat catgtctgga tcgatccagg acctccctaa ccctatgacg
     3601 taattcacgt cacgactcca cccctccagg aacccctagt gatggagttg gccactccct
     3661 ctctgcgcgc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3721 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3781 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3841 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3901 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3961 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4021 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4081 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4141 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4201 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4261 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4321 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4381 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4441 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4501 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4561 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4621 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4681 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4741 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4801 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4861 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4921 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4981 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5041 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5101 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5161 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5221 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5281 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5341 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5401 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5461 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5521 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5581 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5641 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5701 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5761 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5821 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5881 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5941 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6001 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca ttcgccattc
     6061 aggctgcgca actgttggga agggcgatcg
//
