LOCUS       sequence-324257-        6451 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 6451)
  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 6451)
  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..6451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     605..624
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1035..1364)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1049..1184
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1349..1369
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1397..1645
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1397..1417
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1568..1587
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     polyA_signal    2090..2211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   2260..2389
                     /label=AAV2 ITR
     rep_origin      complement(2520..2948)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2630..2651)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2842..2861
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2975..3079
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3080..3148,3149..3940)
                     /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(3298..3317)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4111..4699
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4600..4619
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5142..5276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5701..5721)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5851..5916
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     primer_bind     complement(join(6448..6451,1..16))
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
       61 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
      121 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
      181 ggcgcgagcc cctgatgctc ttgtccagat catcctgatc gacaagaccg gcttccatcc
      241 gagtacgtgc tcgctcgatg cgatgttcgc ttggtggtcg aatgggcagg tagccggatc
      301 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
      361 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
      421 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
      481 ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
      541 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
      601 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
      661 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc
      721 cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc
      781 aaccttacca gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc
      841 ccagtctagc tatcggcatg taagcccact gcaagctacc tgctttctct ttgcgcttgc
      901 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
      961 cggactggct ttctacgtgt tccgcttcct ttagcagccc ttgcgccctg agtgcttgcg
     1021 gcagcgtgaa gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     1081 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     1141 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     1201 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     1261 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     1321 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagccggatc
     1381 caaggtcggg caggaagagg gcctatttcc catgattcct tcatatttgc atatacgata
     1441 caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca
     1501 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     1561 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     1621 tatcttgtgg aaaggacgaa acaccgtgct cgcttcggca gcacatatac tagtgattga
     1681 tgggttgcgg ctttctgctg caatccgacg gccatcagtc gccggtccca agcccggata
     1741 aaatgggagg gggcgggaaa ccgcctaacc atgccgactg atggcaggcc cggatagctc
     1801 agtcggtaga gcagcggccg cgcgattcga ggaaggaggt ctgaggaggt cactgaatcg
     1861 cgcggccgcg ggtccagggt tcaagtccct gttcgggcct gccatcagtc ggcgtggact
     1921 gtagaacact gccaatgccg gtcccaagcc cggataaaag tggagggtac agtccacgct
     1981 ctagagcgga cttcggtccg ctttttacta ggacctgcag gcatgcaagc ttgacgtcgg
     2041 ttaccgatat ccatatggcg gccgcatcga tctcgagccg cggactagta acttgtttat
     2101 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2161 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctt
     2221 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     2281 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     2341 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcaga gagggagtgg
     2401 ccaaagatct ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2461 gcagcctgaa tggctaatgg gaaattgtaa acgttaatat tttgttaata ttttgttaaa
     2521 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     2581 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     2641 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     2701 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     2761 taaagcacta aatcggaacc ctaaagggat gccccgattt agagcttgac ggggaaagcc
     2821 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     2881 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     2941 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3001 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3061 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3121 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3181 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3241 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3301 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3361 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3421 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3481 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3541 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3601 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3661 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3721 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3781 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3841 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3901 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3961 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4021 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4081 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4141 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4201 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     4261 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4321 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4381 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4441 gtgcaacaca gccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4501 gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4561 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4621 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4681 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4741 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4801 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4861 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     4921 gattcattaa tgcagagatc tttggccact ccctctctgc gcgctcgctc gctcactgag
     4981 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     5041 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctggag gggtggagtc
     5101 gtgacgtgaa ttacgtcata gggttaggga ggtcctggat cgatccagac atgataagat
     5161 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     5221 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     5281 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     5341 gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tctctagtca aggcactata
     5401 catcaaatat tccttattaa cccctttaca aattaaaaag ctaaaggtac acaatttttg
     5461 agcatagtta ttaatagcag acactctatg cctgtgtgga gtaagaaaaa acagtatgtt
     5521 atgattataa ctgttatgcc tacttataaa ggttacagaa tatttttcca taattttctt
     5581 gtatagcagt gcagcttttt cctttgtggt gtaaatagca aagcaagcaa gagttctatt
     5641 actaaacaca gcatgactca aaaaacttag caattctgaa ggaaagtcct tggggtcttc
     5701 tacctttctc ttcttttttg gaggagtaga atgttgagag tcagcagtag cctcatcatc
     5761 actagatggc atttcttctg agcaaaacag gttttcctca ttaaaggcat tccaccactg
     5821 ctcccattca tcagttccat aggttggaat ctaaaataca caaacaatta gaatcagtag
     5881 tttaacacat tatacactta aaaattttat atttacctta gagctttaaa tctctgtagg
     5941 tagtttgtcc aattatgtca caccacagaa gtaaggttcc ttcacaaaga tccgggacca
     6001 aagcggccat cgtgcctccc cactcctgca gttcgggggc atggatgcgc ggatagccgc
     6061 tgctggtttc ctggatgccg acggatttgc actgccggta gaactccgcg aggtcgtcca
     6121 gcctcaggca gcagctgaac caactcgcga ggggatcgag cccggggtgg gcgaagaact
     6181 ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga
     6241 agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg
     6301 tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     6361 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     6421 agcccattcg ccgccaagct cttcagcaat a
//
