LOCUS       sequence-330476-        6241 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 6241)
  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 6241)
  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+27-Linear-1, PolyU(0)
FEATURES             Location/Qualifiers
     source          1..6241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..330)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      15..150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     315..335
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        363..611
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     363..383
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     534..553
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    1029..1150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   1199..1328
                     /label=AAV2 ITR
     rep_origin      complement(1422..1877)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1559..1580)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1771..1790
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1904..2008
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2009..2077,2078..2869)
                     /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(2227..2246)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3040..3628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3529..3548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3897..4031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4456..4476)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4606..4671
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(5091..5885)
                     /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(5202..5221)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5812..5831
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
       61 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
      121 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
      181 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
      241 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
      301 tggggacttt ccacacccta actgacacac attccacagc cggatccaag gtcgggcagg
      361 aagagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
      421 agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
      481 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
      541 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
      601 gacgaaacac cgtgctcgct tcggcagcac atatactagt tggccgacgg ccatcagtcg
      661 ccggtcccaa gcccggataa aatgggaggg ggcgggaaac cgcctaacca tgccgactga
      721 tggcaggccc ggatagctca gtcggtagag cagcggccgc gcgattcgag gaaggaggtc
      781 tgaggaggtc actgaatcgc gcggccgcgg gtccagggtt caagtccctg ttcgggcctg
      841 ccatcagtcg gcgtggactg tagaacactg ccaatgccgg tcccaagccc ggataaaagt
      901 ggagggtaca gtccacgctc tagagcggac ttcggtccgc tttttactag gacctgcagg
      961 catgcaagct tgacgtcggt taccgatatc catatggcgg ccgcatcgat ctcgagccgc
     1021 ggactagtaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     1081 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     1141 atgtatctta tcatgtctta cgtagataag tagcatggcg ggttaatcat taactacaag
     1201 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     1261 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     1321 gcgcgcagag agggacagat ctgcagatct gctggcgtaa tagcgaagag gcccgcaccg
     1381 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaaattgtaa gcgttaatat
     1441 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     1501 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     1561 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     1621 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     1681 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     1741 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     1801 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     1861 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1921 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     1981 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2041 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2101 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2161 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2221 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     2281 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2341 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2401 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2461 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2521 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2581 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2641 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2701 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2761 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2821 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2881 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     2941 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3001 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3061 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3121 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3181 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3241 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3301 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3361 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3421 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3481 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3541 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3601 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3661 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3721 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3781 cagcgagtca gtgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
     3841 tggaggggtg gagtcgtgac gtgaattacg tcatagggtt agggaggtcc tggatcgatc
     3901 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     3961 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     4021 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     4081 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatctct
     4141 agtcaaggca ctatacatca aatattcctt attaacccct ttacaaatta aaaagctaaa
     4201 ggtacacaat ttttgagcat agttattaat agcagacact ctatgcctgt gtggagtaag
     4261 aaaaaacagt atgttatgat tataactgtt atgcctactt ataaaggtta cagaatattt
     4321 ttccataatt ttcttgtata gcagtgcagc tttttccttt gtggtgtaaa tagcaaagca
     4381 agcaagagtt ctattactaa acacagcatg actcaaaaaa cttagcaatt ctgaaggaaa
     4441 gtccttgggg tcttctacct ttctcttctt ttttggagga gtagaatgtt gagagtcagc
     4501 agtagcctca tcatcactag atggcatttc ttctgagcaa aacaggtttt cctcattaaa
     4561 ggcattccac cactgctccc attcatcagt tccataggtt ggaatctaaa atacacaaac
     4621 aattagaatc agtagtttaa cacattatac acttaaaaat tttatattta ccttagagct
     4681 ttaaatctct gtaggtagtt tgtccaatta tgtcacacca cagaagtaag gttccttcac
     4741 aaagatccgg accaaagcgg ccatcgtgcc tccccactcc tgcagttcgg gggcatggat
     4801 gcgcggatag ccgctgctgg tttcctggat gccgacggat ttgcactgcc ggtagaactc
     4861 cgcgaggtcg tccagcctca ggcagcagct gaaccaactc gcgaggggat cgagcccggg
     4921 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
     4981 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga
     5041 tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac
     5101 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     5161 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     5221 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     5281 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
     5341 tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     5401 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     5461 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     5521 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     5581 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     5641 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     5701 tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     5761 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     5821 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     5881 atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag
     5941 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     6001 ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat
     6061 cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc
     6121 cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc
     6181 tacgtgttcc gcttccttta gcagcccttg cgccctgagt gcttgcggca gcgtgaagct
     6241 t
//
