LOCUS       sequence-329690-        6245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tests for the impact of a synthetic linear tract 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 6245)
  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 6245)
  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)
FEATURES             Location/Qualifiers
     source          1..6245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(103..558)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(240..261)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     452..471
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        585..689
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(690..758,759..1550)
                     /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(908..927)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1721..2309
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2210..2229
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2578..2712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3137..3157)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          3287..3352
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(3772..4566)
                     /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(3883..3902)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4493..4512
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4923..5252)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4937..5072
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5237..5257
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5285..5533
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5285..5305
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5456..5475
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    5955..6076
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   join(6125..6245,1..9)
                     /label=AAV2 ITR
ORIGIN
        1 agcgcgcaga gagggacaga tctgcagatc tgctggcgta atagcgaaga ggcccgcacc
       61 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggaaattgta agcgttaata
      121 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
      181 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
      241 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
      301 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
      361 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
      421 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
      481 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
      541 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
      601 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      661 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
      721 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
      781 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
      841 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
      901 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
      961 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1021 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1081 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1141 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1201 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     1261 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1321 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     1381 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     1441 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     1501 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     1561 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1621 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1681 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1741 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1801 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1861 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1921 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     1981 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2041 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2101 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2161 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2221 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2281 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2341 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2401 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2461 gcagcgagtc agtgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
     2521 ctggaggggt ggagtcgtga cgtgaattac gtcatagggt tagggaggtc ctggatcgat
     2581 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2641 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2701 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     2761 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatctc
     2821 tagtcaaggc actatacatc aaatattcct tattaacccc tttacaaatt aaaaagctaa
     2881 aggtacacaa tttttgagca tagttattaa tagcagacac tctatgcctg tgtggagtaa
     2941 gaaaaaacag tatgttatga ttataactgt tatgcctact tataaaggtt acagaatatt
     3001 tttccataat tttcttgtat agcagtgcag ctttttcctt tgtggtgtaa atagcaaagc
     3061 aagcaagagt tctattacta aacacagcat gactcaaaaa acttagcaat tctgaaggaa
     3121 agtccttggg gtcttctacc tttctcttct tttttggagg agtagaatgt tgagagtcag
     3181 cagtagcctc atcatcacta gatggcattt cttctgagca aaacaggttt tcctcattaa
     3241 aggcattcca ccactgctcc cattcatcag ttccataggt tggaatctaa aatacacaaa
     3301 caattagaat cagtagttta acacattata cacttaaaaa ttttatattt accttagagc
     3361 tttaaatctc tgtaggtagt ttgtccaatt atgtcacacc acagaagtaa ggttccttca
     3421 caaagatccg gaccaaagcg gccatcgtgc ctccccactc ctgcagttcg ggggcatgga
     3481 tgcgcggata gccgctgctg gtttcctgga tgccgacgga tttgcactgc cggtagaact
     3541 ccgcgaggtc gtccagcctc aggcagcagc tgaaccaact cgcgagggga tcgagcccgg
     3601 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     3661 ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg
     3721 atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa
     3781 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     3841 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     3901 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     3961 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc
     4021 ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     4081 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
     4141 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     4201 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     4261 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     4321 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     4381 gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     4441 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     4501 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     4561 aatcatgcga aacgatcctc atcctgtctc ttgatcagat cttgatcccc tgcgccatca
     4621 gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga
     4681 gggcgcccca gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta
     4741 tcgccatgta agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt
     4801 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     4861 ctacgtgttc cgcttccttt agcagccctt gcgccctgag tgcttgcggc agcgtgaagc
     4921 tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag
     4981 gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa
     5041 tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt
     5101 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     5161 ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     5221 cctggggact ttccacaccc taactgacac acattccaca gccggatcca aggtcgggca
     5281 ggaagagggc ctatttccca tgattccttc atatttgcat atacgataca aggctgttag
     5341 agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
     5401 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
     5461 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
     5521 aggacgaaac accgtgctcg cttcggcagc acatatacta gtgattgatg cgacggccat
     5581 cagtcgccgg tcccaagccc ggataaaatg ggagggggcg ggaaaccgcc taaccatgcc
     5641 gactgatggc aggcccggat agctcagtcg gtagagcagc ggccgcgcga ttcgaggaag
     5701 gaggtctgag gaggtcactg aatcgcgcgg ccgcgggtcc agggttcaag tccctgttcg
     5761 ggcctgccat cagtcggcgt ggactgtaga acactgccaa tgccggtccc aagcccggat
     5821 aaaagtggag ggtacagtcc acgctctaga gcggacttcg gtccgctttt tactaggacc
     5881 tgcaggcatg caagcttgac gtcggttacc gatatccata tggcggccgc atcgatctcg
     5941 agccgcggac tagtaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6001 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6061 tcatcaatgt atcttatcat gtcttacgta gataagtagc atggcgggtt aatcattaac
     6121 tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     6181 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     6241 gagcg
//
