LOCUS       sequence-329417-        6264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of a synthetic linear tract in conjunction with
            a hairpin motif on human polymerase III transcription from a U6 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6264)
  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 6264)
  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: pAV-U6+27Linear-1, PolyU(0), Hairpin Distance ...
FEATURES             Location/Qualifiers
     source          1..6264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..250
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2..22
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     173..192
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    691..812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   861..990
                     /label=AAV2 ITR
     rep_origin      complement(1084..1539)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1221..1242)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1433..1452
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1566..1670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1671..1739,1740..2531)
                     /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(1889..1908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2702..3290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3191..3210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3559..3693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4118..4138)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4268..4333
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(4753..5547)
                     /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(4864..4883)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5474..5493
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5904..6233)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5918..6053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6218..6238
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
       61 gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
      121 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
      181 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
      241 acgaaacacc gtgctcgctt cggcagcaca tatactagtt ggcggttgcg gctttctgct
      301 gcaatccgac ggccatcagt cgccggtccc aagcccggat aaaatgggag ggggcgggaa
      361 accgcctaac catgccgact gatggcaggc ccggatagct cagtcggtag agcagcggcc
      421 gcgcgattcg aggaaggagg tctgaggagg tcactgaatc gcgcggccgc gggtccaggg
      481 ttcaagtccc tgttcgggcc tgccatcagt cggcgtggac tgtagaacac tgccaatgcc
      541 ggtcccaagc ccggataaaa gtggagggta cagtccacgc tctagagcgg acttcggtcc
      601 gctttttact aggacctgca ggcatgcaag cttgacgtcg gttaccgata tccatatggc
      661 ggccgcatcg atctcgagcc gcggactagt aacttgttta ttgcagctta taatggttac
      721 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
      781 tgtggtttgt ccaaactcat caatgtatct tatcatgtct tacgtagata agtagcatgg
      841 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
      901 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
      961 ggcggcctca gtgagcgagc gagcgcgcag agagggacag atctgcagat ctgctggcgt
     1021 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     1081 tggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     1141 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     1201 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     1261 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     1321 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     1381 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     1441 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     1501 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     1561 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1621 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1681 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1741 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1801 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1861 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     1921 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1981 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2041 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2101 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2161 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2221 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2281 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2341 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2401 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2461 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2521 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2581 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2641 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2701 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2761 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2821 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2881 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2941 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3001 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3061 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3121 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3181 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3241 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3301 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3361 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3421 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga gcgcgcagag agggagtggc
     3481 caactccatc actaggggtt cctggagggg tggagtcgtg acgtgaatta cgtcataggg
     3541 ttagggaggt cctggatcga tccagacatg ataagataca ttgatgagtt tggacaaacc
     3601 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3661 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3721 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3781 ggtatggctg attatgatct ctagtcaagg cactatacat caaatattcc ttattaaccc
     3841 ctttacaaat taaaaagcta aaggtacaca atttttgagc atagttatta atagcagaca
     3901 ctctatgcct gtgtggagta agaaaaaaca gtatgttatg attataactg ttatgcctac
     3961 ttataaaggt tacagaatat ttttccataa ttttcttgta tagcagtgca gctttttcct
     4021 ttgtggtgta aatagcaaag caagcaagag ttctattact aaacacagca tgactcaaaa
     4081 aacttagcaa ttctgaagga aagtccttgg ggtcttctac ctttctcttc ttttttggag
     4141 gagtagaatg ttgagagtca gcagtagcct catcatcact agatggcatt tcttctgagc
     4201 aaaacaggtt ttcctcatta aaggcattcc accactgctc ccattcatca gttccatagg
     4261 ttggaatcta aaatacacaa acaattagaa tcagtagttt aacacattat acacttaaaa
     4321 attttatatt taccttagag ctttaaatct ctgtaggtag tttgtccaat tatgtcacac
     4381 cacagaagta aggttccttc acaaagatcc ggaccaaagc ggccatcgtg cctccccact
     4441 cctgcagttc gggggcatgg atgcgcggat agccgctgct ggtttcctgg atgccgacgg
     4501 atttgcactg ccggtagaac tccgcgaggt cgtccagcct caggcagcag ctgaaccaac
     4561 tcgcgagggg atcgagcccg gggtgggcga agaactccag catgagatcc ccgcgctgga
     4621 ggatcatcca gccggcgtcc cggaaaaaga ttccgaagcc caacctttca tagaaggcgg
     4681 cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc
     4741 ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     4801 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     4861 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
     4921 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     4981 gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag
     5041 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     5101 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     5161 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     5221 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     5281 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     5341 ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt
     5401 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     5461 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     5521 tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga
     5581 tcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca
     5641 gggcttccca accttaccag agggcgcccc agctggcaat tccggttcgc ttgctgtcca
     5701 taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct gctttctctt
     5761 tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca
     5821 ccgtttctgc ggactggctt tctacgtgtt ccgcttcctt tagcagccct tgcgccctga
     5881 gtgcttgcgg cagcgtgaag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac
     5941 tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt
     6001 agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga
     6061 gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     6121 gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc
     6181 atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac
     6241 agccggatcc aaggtcgggc agga
//
