LOCUS       sequence-299833-        4900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sleeping Beauty self-reporting transposon with puromycin reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4900)
  AUTHORS   Moudgil A, Wilkinson MN, Chen X, He J, Cammack AJ, Vasek MJ, Lagunas
            T Jr, Qi Z, Lalli MA, Guo C, Morris SA, Dougherty JD, Mitra RD
  TITLE     Self-Reporting Transposons Enable Simultaneous Readout of Gene 
            Expression and Transcription Factor Binding in Single Cells.
  JOURNAL   Cell. 2020 Aug 20;182(4):992-1008.e21. doi: 
            10.1016/j.cell.2020.06.037. Epub 2020 Jul 24.
  PUBMED    32710817
REFERENCE   2  (bases 1 to 4900)
  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.cell.2020.06.037"; journalName: "Cell"; date: 
            "2020-08-20- 20"; volume: "182"; issue: "4"; pages: "992-1008.e21"
COMMENT     Sequence Label: SB-SRT-Puro
FEATURES             Location/Qualifiers
     source          1..4900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             21..620
                     /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(21..40)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     517..537
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1282..1299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1392..1980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1472..1491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2151..2942,2943..3011))
                     /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     2774..2793
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3012..3116)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(3725..4900,1..7)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3956..4898
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4855..4875
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 gtcgtgaaaa ctacgccacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
       61 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
      121 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
      181 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
      241 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
      301 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
      361 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
      421 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
      481 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
      541 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
      601 cccgcaagcc cggtgcctga taacaactcg agtaccggct taagtgatag atcggaagag
      661 cggttcagca ggaatgccga gaccggttta aactcagtgt ccatagcgac gccggcgcga
      721 taagctggac cccttaccga aacatcgccg cattctgcag aggagtcgaa cttagtgtat
      781 gtaaacttct gacccactgg aattgtgata cagtgaatta taagtgaaat aatctgtctg
      841 taaacaattg ttggaaaaat gacttgtgtc atgcacaaag tagatgtcct aactgacttg
      901 ccaaaactat tgtttgttaa caagaaattt gtggagtagt tgaaaaacga gttttaatga
      961 ctccaactta agtgtatgta aacttccgac ttcaactgta tagagctcct gagatgcagg
     1021 tacatcccac tgatgagtcc caaataggac gaaacgcgct tcggtgcgtc tgggattcca
     1081 ctgctatcca cgcggccgca cggcattacg ttagacgtct aggccacacc accctgaagg
     1141 cgcctgctcg cctctgatct gttgaagcta agcagggtcg gtcctggtta gtacttggat
     1201 gggactccgc ctggtaatag ccggtaccgt aggcttgcat ggtcatagct gtttccttgc
     1261 gtattgggcg ctctccgctt cctcgctcac tgactcgctg cgctcggtcg ttcgggtaaa
     1321 gcctggggtg cctaatgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1381 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1441 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1501 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1561 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1621 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1681 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1741 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1801 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1861 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1921 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1981 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2041 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2101 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2161 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2221 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2281 atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2341 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     2401 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     2461 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     2521 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     2581 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2641 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2701 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     2761 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     2821 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2881 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     2941 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3001 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     3061 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     3121 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3181 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3241 aatcaaaaga atagaccgag atagggttga gtggccgcta cagggcgctc ccattcgcca
     3301 ttcaggctgc gcaactgttg ggaagggcgt ttcggtgcgg gcctcttcgc tattacgcca
     3361 gctgggcctt ttgctggcct tttgctctat acagttgaag tcggaagttt acatacacct
     3421 tagccaaata catttaaact cactttttca caattcctga catttaatcc tagtaaaaat
     3481 tccctgtctt aggtcagtta ggatcaccac tttattttaa gaatgtgaaa tatcagaata
     3541 atagtagaga gaatgattca tttcagcttt tatttctttc atcacattcc cagtgggtca
     3601 gaagtttaca tacactcgac tcctctgcag aatgcggcga tgtttcggta aggggtcctt
     3661 aattaatacc acatttgtag aggttttact tgctttaaaa aactctagag aagagcatgc
     3721 gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     3781 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga
     3841 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     3901 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
     3961 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
     4021 aattacttcc acgcccctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa
     4081 gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg
     4141 aggcctggct tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc
     4201 tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt
     4261 ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt
     4321 ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg
     4381 gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc
     4441 tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt
     4501 cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa
     4561 aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg
     4621 cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc
     4681 acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt
     4741 atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca gcttggcact
     4801 tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc
     4861 ctcagacagt ggttcaaagt ttttttcttc catttcaggt
//
