LOCUS       sequence-173710-        6813 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  used for downstream cloning, contains CMV promoter with NheI and 
            XhoI cloning sites flanking a partial insert that gets excised when 
            adding an insert of choice.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6813)
  AUTHORS   Goldfarb KC, Cech TR
  TITLE     Targeted CRISPR disruption reveals a role for RNase MRP RNA in human
            preribosomal RNA processing.
  JOURNAL   Genes Dev. 2017 Jan 1;31(1):59-71. doi: 10.1101/gad.286963.116. Epub
            2017 Jan 23.
  PUBMED    28115465
REFERENCE   2  (bases 1 to 6813)
  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"; doi: 
            "10.1101/gad.286963.116"; journalName: "Genes Dev"; date: 
            "2017-01-1- 1"; volume: "31"; issue: "1"; pages: "59-71"
COMMENT     Sequence Label: psglessClonePX
FEATURES             Location/Qualifiers
     source          1..6813
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     polyA_signal    904..952
                     /note="synthetic polyadenylation signal"
     protein_bind    complement(1008..1041)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1097..1685
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1150..1170)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1568..1579)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1688..1704)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1689..1705)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2214..2394
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(2456..3044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2536..2555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3215..4006,4007..4075))
                     /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     3838..3857
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4076..4180)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4255..4274)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4446..4825
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4827..5025
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4980..5000
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5043..5223
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5270..5395
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5892..6125
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6310..6354
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    6652..6769
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgaagt tagtgataat
       61 gggattggtt ctgatattga gctgcttaga gcagctgttt tgaaaaatgg gggtgggtgg
      121 catggtcttg ggtgggttgg agaaggcaaa tggattgtga agaaaggtaa tgttagctca
      181 gccgggaaat gtttgagctg tgatgagcat ttggcttgtg ttgataccaa tgaggtagag
      241 actgaagatt ttgtgaattc tttggtgact ttggctatgg agaggaaggc aaagatgaat
      301 tcatgcgagc ctatggcgga tttcagtgag tttcaggagt ggcttgaaaa gcatggagat
      361 tacgaagcta tactagatgg agcaaatatt gggctctacc aacaaaattt tgcagatggt
      421 ggtttcagtc taccacagct tgaagctgta gtgaaggaac tttacaataa aagtggtagc
      481 aaaaaacagc cgctaattct cttgcacaag aaacgggtca atgcgctttt agaaaacccg
      541 aatcaccgga atcttgtgga agaatggatt aataacaatg tcctgtatgc gactccacca
      601 ggttccaatg atgattggta ttggctctac gctgctgcta aactcaagtg tttacttgtg
      661 actaatgatg agatgagaga tcacattttt gaactattaa gtaacagttt cttccaaaaa
      721 tggaaagaaa gacatcaagt tcggtttacc tttgtgaaag gctgtctcaa gcttgaaatg
      781 ccacctcctt tttcagttgt gatccaggaa tcggagaaag gatcatggca cgttccaatc
      841 acgagtcaag acaaggagga atctttaaga agttggatgt gcattacaag gcagagttca
      901 tagaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgtatagcat
      961 acattatacg aagttatgga tcctcgagca attcgtcgag ggacctaata acttcgtata
     1021 gcatacatta tacgaagtta tacatgttta agggttccgg ttccactagg tacaattcga
     1081 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1141 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1201 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     1261 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     1321 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     1381 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     1441 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     1501 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     1561 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     1621 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     1681 cccgcatcga taccgtcgac ctcgatcgag acctagaaaa acatggagca atcacaagta
     1741 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
     1801 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
     1861 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     1921 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga
     1981 actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc tacaagctag
     2041 taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc cgcttgttac
     2101 accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag tggaggtttg
     2161 acagccgcct agcatttcat cacatggccc gagagctgca tccggactgt actgggtctc
     2221 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     2281 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     2341 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagcatgt
     2401 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2461 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2521 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2581 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2641 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2701 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2761 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2821 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2881 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2941 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3001 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3061 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3121 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3181 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3241 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3301 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3361 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3421 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3481 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3541 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3601 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3661 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3721 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3781 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3841 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3901 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3961 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4021 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4081 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4141 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4201 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     4261 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     4321 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     4381 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     4441 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     4501 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     4561 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     4621 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     4681 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     4741 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     4801 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     4861 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     4921 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     4981 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta
     5041 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5101 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     5161 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     5221 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     5281 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     5341 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     5401 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     5461 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     5521 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     5581 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     5641 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     5701 gcaaaacaaa agtaagacca ccgcacagca agcggccggc cgctgatctt cagacctgga
     5761 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     5821 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     5881 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     5941 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     6001 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     6061 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     6121 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     6181 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     6241 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     6301 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     6361 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     6421 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     6481 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
     6541 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
     6601 tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag
     6661 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     6721 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     6781 cagggacagc agagatccag tttggttagt acc
//
