LOCUS       sequence-330472-        6089 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 GLY tRNA promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6089)
  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 6089)
  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), Hairpin Distance = 0nt
FEATURES             Location/Qualifiers
     source          1..6089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..21)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          151..216
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(636..1430)
                     /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(747..766)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1357..1376
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1787..2116)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1801..1936
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2101..2121
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    2663..2784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2833..2962
                     /label=AAV2 ITR
     rep_origin      complement(3056..3511)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3193..3214)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3405..3424
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3538..3642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3643..3711,3712..4503)
                     /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(3861..3880)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4674..5262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5163..5182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5531..5665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tacctttctc ttcttttttg gaggagtaga atgttgagag tcagcagtag cctcatcatc
       61 actagatggc atttcttctg agcaaaacag gttttcctca ttaaaggcat tccaccactg
      121 ctcccattca tcagttccat aggttggaat ctaaaataca caaacaatta gaatcagtag
      181 tttaacacat tatacactta aaaattttat atttacctta gagctttaaa tctctgtagg
      241 tagtttgtcc aattatgtca caccacagaa gtaaggttcc ttcacaaaga tccggaccaa
      301 agcggccatc gtgcctcccc actcctgcag ttcgggggca tggatgcgcg gatagccgct
      361 gctggtttcc tggatgccga cggatttgca ctgccggtag aactccgcga ggtcgtccag
      421 cctcaggcag cagctgaacc aactcgcgag gggatcgagc ccggggtggg cgaagaactc
      481 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
      541 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
      601 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
      661 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
      721 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
      781 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
      841 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
      901 cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
      961 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     1021 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     1081 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     1141 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     1201 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc
     1261 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     1321 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     1381 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     1441 cctcatcctg tctcttgatc agatcttgat cccctgcgcc atcagatcct tggcggcaag
     1501 aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc
     1561 aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca
     1621 ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag
     1681 ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt gttccgcttc
     1741 ctttagcagc ccttgcgccc tgagtgcttg cggcagcgtg aagctttttg caaaagccta
     1801 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     1861 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     1921 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     1981 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     2041 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     2101 accctaactg acacacattc cacagccgga tccaaggtcg ggcaggaagt ctctcgtcgc
     2161 attggtggtt cagtggtaga attctcgcct gccacgcggg aggcccgggt tcgattcccg
     2221 gccaatgcaa tatactagca gccaactcca aggttgcggc tttctgctgc aatcttttcg
     2281 acggccatca gtcgccggtc ccaagcccgg ataaaatggg agggggcggg aaaccgccta
     2341 accatgccga ctgatggcag gcccggatag ctcagtcggt agagcagcgg ccgcgcgatt
     2401 cgaggaagga ggtctgagga ggtcactgaa tcgcgcggcc gcgggtccag ggttcaagtc
     2461 cctgttcggg cctgccatca gtcggcgtgg actgtagaac actgccaatg ccggtcccaa
     2521 gcccggataa aagtggaggg tacagtccac gctctagagc ggacttcggt ccgcttttta
     2581 ctaggacctg caggcatgca agcttgacgt cggttaccga tatccatatg gcggccgcat
     2641 cgatctcgag ccgcggacta gtaacttgtt tattgcagct tataatggtt acaaataaag
     2701 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2761 gtccaaactc atcaatgtat cttatcatgt cttacgtaga taagtagcat ggcgggttaa
     2821 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     2881 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     2941 cagtgagcga gcgagcgcgc agagagggac agatctgcag atctgctggc gtaatagcga
     3001 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggaaatt
     3061 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3121 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3181 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3241 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3301 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3361 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3421 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3481 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3541 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3601 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3661 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3721 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3781 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3841 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3901 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3961 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4021 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4081 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4141 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4201 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4261 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4321 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4381 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4441 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4501 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4561 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4621 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4681 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4741 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4801 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4861 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4921 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4981 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5041 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5101 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5161 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5221 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5281 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5341 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5401 cgaacgaccg agcgcagcga gtcagtgagc gagcgcgcag agagggagtg gccaactcca
     5461 tcactagggg ttcctggagg ggtggagtcg tgacgtgaat tacgtcatag ggttagggag
     5521 gtcctggatc gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     5581 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     5641 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     5701 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     5761 tgattatgat ctctagtcaa ggcactatac atcaaatatt ccttattaac ccctttacaa
     5821 attaaaaagc taaaggtaca caatttttga gcatagttat taatagcaga cactctatgc
     5881 ctgtgtggag taagaaaaaa cagtatgtta tgattataac tgttatgcct acttataaag
     5941 gttacagaat atttttccat aattttcttg tatagcagtg cagctttttc ctttgtggtg
     6001 taaatagcaa agcaagcaag agttctatta ctaaacacag catgactcaa aaaacttagc
     6061 aattctgaag gaaagtcctt ggggtcttc
//
