LOCUS       sequence-170165-        7409 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7409)
  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 7409)
  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..7409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        95..398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        399..602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     polyA_signal    1489..1537
                     /note="synthetic polyadenylation signal"
     protein_bind    complement(1593..1626)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1682..2270
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1735..1755)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2153..2164)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2273..2289)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2274..2290)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2799..2979
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3041..3629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3121..3140)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3800..4591,4592..4660))
                     /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     4423..4442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4661..4765)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4849..4868)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5040..5419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5421..5619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             5637..5817
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5864..5989
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6488..6721
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6906..6950
                     /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    7248..7365
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gggccgcgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      601 ctggtttagt gaaccgtcag atccgctagc gaagttagtg ataatgggat tggttctgat
      661 attgagctgc ttagagcagc tgttttgaaa aatgggggtg ggtggcatgg tcttgggtgg
      721 gttggagaag gcaaatggat tgtgaagaaa ggtaatgtta gctcagccgg gaaatgtttg
      781 agctgtgatg agcatttggc ttgtgttgat accaatgagg tagagactga agattttgtg
      841 aattctttgg tgactttggc tatggagagg aaggcaaaga tgaattcatg cgagcctatg
      901 gcggatttca gtgagtttca ggagtggctt gaaaagcatg gagattacga agctatacta
      961 gatggagcaa atattgggct ctaccaacaa aattttgcag atggtggttt cagtctacca
     1021 cagcttgaag ctgtagtgaa ggaactttac aataaaagtg gtagcaaaaa acagccgcta
     1081 attctcttgc acaagaaacg ggtcaatgcg cttttagaaa acccgaatca ccggaatctt
     1141 gtggaagaat ggattaataa caatgtcctg tatgcgactc caccaggttc caatgatgat
     1201 tggtattggc tctacgctgc tgctaaactc aagtgtttac ttgtgactaa tgatgagatg
     1261 agagatcaca tttttgaact attaagtaac agtttcttcc aaaaatggaa agaaagacat
     1321 caagttcggt ttacctttgt gaaaggctgt ctcaagcttg aaatgccacc tcctttttca
     1381 gttgtgatcc aggaatcgga gaaaggatca tggcacgttc caatcacgag tcaagacaag
     1441 gaggaatctt taagaagttg gatgtgcatt acaaggcaga gttcatagaa taaaatatct
     1501 ttattttcat tacatctgtg tgttggtttt ttgtgtgtat agcatacatt atacgaagtt
     1561 atggatcctc gagcaattcg tcgagggacc taataacttc gtatagcata cattatacga
     1621 agttatacat gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga
     1681 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1741 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     1801 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     1861 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     1921 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     1981 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2041 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     2101 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2161 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2221 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     2281 tcgacctcga tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
     2341 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
     2401 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     2461 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
     2521 atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc
     2581 cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag
     2641 agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc
     2701 atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat
     2761 ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga
     2821 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     2881 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     2941 ccctcagacc cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc
     3001 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3061 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3121 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3181 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3241 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3301 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3361 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3421 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3481 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcttaag
     4801 cggtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     4861 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     4921 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     4981 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
     5041 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     5101 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     5161 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     5221 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     5281 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     5341 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     5401 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     5461 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     5521 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     5581 gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt
     5641 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5701 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     5761 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     5821 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     5881 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     5941 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     6001 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     6061 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     6121 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     6181 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     6241 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     6301 caaaagtaag accaccgcac agcaagcggc cggccgcgct gatcttcaga cctggaggag
     6361 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     6421 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     6481 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     6541 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     6601 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     6661 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     6721 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     6781 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     6841 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     6901 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     6961 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     7021 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     7081 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     7141 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg
     7201 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa
     7261 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     7321 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     7381 gacagcagag atccagtttg gttagtacc
//
