LOCUS       sequence-329223-        6268 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 6268)
  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 6268)
  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-2, PolyU(0), Hairpin Distance ...
FEATURES             Location/Qualifiers
     source          1..6268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(54..509)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(191..212)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     403..422
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        536..640
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(641..709,710..1501)
                     /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(859..878)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1672..2260
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2161..2180
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2529..2663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3088..3108)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          3238..3303
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(3723..4517)
                     /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(3834..3853)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4444..4463
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4874..5203)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4888..5023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5188..5208
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5236..5484
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5236..5256
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5407..5426
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    5929..6050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   6099..6228
                     /label=AAV2 ITR
ORIGIN
        1 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt
       61 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
      121 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
      181 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
      241 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
      301 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
      361 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
      421 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
      481 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
      541 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      601 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
      661 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
      721 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
      781 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
      841 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
      901 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
      961 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     1021 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     1081 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     1141 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     1201 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     1261 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     1321 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     1381 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     1441 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1501 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     1561 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     1621 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1681 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1741 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1801 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1861 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1921 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1981 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2041 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2101 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2161 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2221 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2281 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2341 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2401 aacgaccgag cgcagcgagt cagtgagcga gcgcgcagag agggagtggc caactccatc
     2461 actaggggtt cctggagggg tggagtcgtg acgtgaatta cgtcataggg ttagggaggt
     2521 cctggatcga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2581 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2641 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2701 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     2761 attatgatct ctagtcaagg cactatacat caaatattcc ttattaaccc ctttacaaat
     2821 taaaaagcta aaggtacaca atttttgagc atagttatta atagcagaca ctctatgcct
     2881 gtgtggagta agaaaaaaca gtatgttatg attataactg ttatgcctac ttataaaggt
     2941 tacagaatat ttttccataa ttttcttgta tagcagtgca gctttttcct ttgtggtgta
     3001 aatagcaaag caagcaagag ttctattact aaacacagca tgactcaaaa aacttagcaa
     3061 ttctgaagga aagtccttgg ggtcttctac ctttctcttc ttttttggag gagtagaatg
     3121 ttgagagtca gcagtagcct catcatcact agatggcatt tcttctgagc aaaacaggtt
     3181 ttcctcatta aaggcattcc accactgctc ccattcatca gttccatagg ttggaatcta
     3241 aaatacacaa acaattagaa tcagtagttt aacacattat acacttaaaa attttatatt
     3301 taccttagag ctttaaatct ctgtaggtag tttgtccaat tatgtcacac cacagaagta
     3361 aggttccttc acaaagatcc ggaccaaagc ggccatcgtg cctccccact cctgcagttc
     3421 gggggcatgg atgcgcggat agccgctgct ggtttcctgg atgccgacgg atttgcactg
     3481 ccggtagaac tccgcgaggt cgtccagcct caggcagcag ctgaaccaac tcgcgagggg
     3541 atcgagcccg gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca
     3601 gccggcgtcc cggaaaaaga ttccgaagcc caacctttca tagaaggcgg cggtggaatc
     3661 gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc
     3721 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     3781 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     3841 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     3901 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     3961 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
     4021 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     4081 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     4141 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     4201 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     4261 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     4321 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     4381 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     4441 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     4501 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc
     4561 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     4621 accttaccag agggcgcccc agctggcaat tccggttcgc ttgctgtcca taaaaccgcc
     4681 cagtctagct atcgccatgt aagcccactg caagctacct gctttctctt tgcgcttgcg
     4741 ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc
     4801 ggactggctt tctacgtgtt ccgcttcctt tagcagccct tgcgccctga gtgcttgcgg
     4861 cagcgtgaag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga
     4921 atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat
     4981 ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg
     5041 ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     5101 ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc
     5161 ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac agccggatcc
     5221 aaggtcgggc aggaagaggg cctatttccc atgattcctt catatttgca tatacgatac
     5281 aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa
     5341 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
     5401 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
     5461 atcttgtgga aaggacgaaa caccgtgctc gcttcggcag cacatatact agtgattgat
     5521 gggttgcggc tttctgctgc aatccgacgg ccatcagtcg ccggtcccaa gcccggataa
     5581 aatgggaggg ggcgggaaac cgcctaacca tgccgactga tggcaggccc ggatagctca
     5641 gtcggtagag cagcggccgc gcgattcgag gaaggaggtc tgaggaggtc actgaatcgc
     5701 gcggccgcgg gtccagggtt caagtccctg ttcgggcctg ccatcagtcg gcgtggactg
     5761 tagaacactg ccaatgccgg tcccaagccc ggataaaagt ggagggtaca gtccacgctc
     5821 tagagcggac ttcggtccgc tttttactag gacctgcagg catgcaagct tgacgtcggt
     5881 taccgatatc catatggcgg ccgcatcgat ctcgagccgc ggactagtaa cttgtttatt
     5941 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6001 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctta
     6061 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     6121 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     6181 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagag agggacagat
     6241 ctgcagatct gctggcgtaa tagcgaag
//
