LOCUS       sequence-173469-        1784 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cloning backbone for sgRNA, also has a barcode that allows detection
            from scRNA-seq.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1784)
  AUTHORS   Xie S, Duan J, Li B, Zhou P, Hon GC
  TITLE     Multiplexed Engineering and Analysis of Combinatorial Enhancer 
            Activity in Single Cells.
  JOURNAL   Mol Cell. 2017 Apr 20;66(2):285-299.e5. doi: 
            10.1016/j.molcel.2017.03.007. Epub 2017 Apr 13.
  PUBMED    28416141
REFERENCE   2  (bases 1 to 1784)
  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: 
            "10.1016/j.molcel.2017.03.007"; journalName: "Mol Cell"; date: 
            "2017-04-20- 20"; volume: "66"; issue: "2"; pages: "285-299.e5"
COMMENT     Sequence Label: Lenti-sgRNA(MS2)-puro-barcode
FEATURES             Location/Qualifiers
     source          1..1784
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        13..224
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          225..1163
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1120..1140
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1185..1784
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1185..1204)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1681..1701
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
       61 tcggcaattg atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
      121 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
      181 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg
      241 gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca
      301 cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt
      361 cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc
      421 gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata
      481 agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata
      541 gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg
      601 gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc
      661 caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg
      721 cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt
      781 gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc
      841 gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag
      901 ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct
      961 cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc
     1021 cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct
     1081 tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc
     1141 aaagtttttt tcttccattt caggtgtcgt gacgtacggc caccatgacc gagtacaagc
     1201 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     1261 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     1321 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     1381 tcgcggacga cggcgccgcc gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     1441 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     1501 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     1561 ccaccgtcgg agtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     1621 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     1681 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     1741 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctga
//
