LOCUS       sequence-112015-        3583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Puromycin-based homologous recombination-dependent reporter for 
            genome editing with TALENs or CRISPR/Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3583)
  AUTHORS   Flemr M, Buhler M
  TITLE     Single-Step Generation of Conditional Knockout Mouse Embryonic Stem 
            Cells.
  JOURNAL   Cell Rep. 2015 Jul 28;12(4):709-16. doi: 
            10.1016/j.celrep.2015.06.051. Epub 2015 Jul 16.
  PUBMED    26190102
REFERENCE   2  (bases 1 to 3583)
  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.celrep.2015.06.051"; journalName: "Cell Rep"; date: 
            "2015-07-28- 28"; volume: "12"; issue: "4"; pages: "709-16"
COMMENT     Sequence Label: pMB1610_pRR-Puro
FEATURES             Location/Qualifiers
     source          1..3583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             918..1514
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     1411..1431
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1538..1659
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1549..1568)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1603..1622
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1792..1896
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1897..1965,1966..2757)
                     /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(2115..2134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2928..3516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3417..3436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcc accatgaccg agtacaagcc cacggtgcgc
      721 ctcgccaccc gcgacgatgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
      781 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
      841 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcgtaatga
      901 gagctcttga cgtctaaacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgatg
      961 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     1021 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     1081 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     1141 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     1201 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     1261 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     1321 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     1381 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     1441 tcaccgtcac cgccgacgta gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     1501 agcccggtgc ctgagcggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     1561 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     1621 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     1681 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     1741 aaatgtggta aaatcgataa ggatccaggt ggcacttttc ggggaaatgt gcgcggaacc
     1801 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1861 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1921 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1981 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2041 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2101 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2161 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2221 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2281 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2341 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2401 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2461 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2521 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2581 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2641 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2701 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2761 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2821 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2881 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2941 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3001 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3061 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3121 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3181 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3241 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3301 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3361 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3421 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3481 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3541 cggttcctgg ccttttgctg gccttttgct cacatggctc gac
//
