LOCUS       sequence-330477-        6087 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 GLN tRNA promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6087)
  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 6087)
  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(1), Hairpin Distance = 0nt
FEATURES             Location/Qualifiers
     source          1..6087
                     /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(890..910)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        895..1224
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1075..1210
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1581..2375
                     /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(1635..1654)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2245..2264
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     intron          2795..2860
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2990..3010
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3435..3569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3838..4426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3918..3937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4597..5388,5389..5457))
                     /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     5220..5239
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5458..5562)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5589..6044
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5676..5695)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5886..5907
                     /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 gtcgagattg cagcagaaag ccgcaacctt ggagttggct gctagtatat
      781 aggttccacc gagatttgaa ctcggatcgc tggattcaga gtccagagtg ctaaccatta
      841 caccatggaa ccgacgagag acttcctgcc cgaccttgga tccggctgtg gaatgtgtgt
      901 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
      961 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     1021 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     1081 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1141 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     1201 tttggaggcc taggcttttg caaaaagctt cacgctgccg caagcactca gggcgcaagg
     1261 gctgctaaag gaagcggaac acgtagaaag ccagtccgca gaaacggtgc tgaccccgga
     1321 tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg
     1381 tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg
     1441 aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact
     1501 ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga
     1561 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     1621 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     1681 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     1741 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     1801 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     1861 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     1921 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     1981 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     2041 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     2101 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     2161 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     2221 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     2281 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     2341 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     2401 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     2461 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     2521 tctcatgctg gagttcttcg cccaccccgg gctcgatccc ctcgcgagtt ggttcagctg
     2581 ctgcctgagg ctggacgacc tcgcggagtt ctaccggcag tgcaaatccg tcggcatcca
     2641 ggaaaccagc agcggctatc cgcgcatcca tgcccccgaa ctgcaggagt ggggaggcac
     2701 gatggccgct ttggtccgga tctttgtgaa ggaaccttac ttctgtggtg tgacataatt
     2761 ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt taagtgtata
     2821 atgtgttaaa ctactgattc taattgtttg tgtattttag attccaacct atggaactga
     2881 tgaatgggag cagtggtgga atgcctttaa tgaggaaaac ctgttttgct cagaagaaat
     2941 gccatctagt gatgatgagg ctactgctga ctctcaacat tctactcctc caaaaaagaa
     3001 gagaaaggta gaagacccca aggactttcc ttcagaattg ctaagttttt tgagtcatgc
     3061 tgtgtttagt aatagaactc ttgcttgctt tgctatttac accacaaagg aaaaagctgc
     3121 actgctatac aagaaaatta tggaaaaata ttctgtaacc tttataagta ggcataacag
     3181 ttataatcat aacatactgt tttttcttac tccacacagg catagagtgt ctgctattaa
     3241 taactatgct caaaaattgt gtacctttag ctttttaatt tgtaaagggg ttaataagga
     3301 atatttgatg tatagtgcct tgactagaga tcataatcag ccataccaca tttgtagagg
     3361 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3421 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3481 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3541 tcatcaatgt atcttatcat gtctggatcg atccaggacc tccctaaccc tatgacgtaa
     3601 ttcacgtcac gactccaccc ctccaggaac ccctagtgat ggagttggcc actccctctc
     3661 tgcgcgctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3721 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3781 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3841 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3901 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3961 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4021 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4081 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4141 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4201 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4261 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4321 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4381 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4441 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4501 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4561 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4621 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4681 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4741 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4801 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4861 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4921 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4981 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5041 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5101 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5161 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5221 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5281 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5341 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5401 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5461 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5521 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5581 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5641 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5701 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5761 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5821 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5881 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5941 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     6001 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg
     6061 ctgcgcaact gttgggaagg gcgatcg
//
