LOCUS       sequence-308645-        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"
     promoter        245..1427
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          476..1418
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1375..1395
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1441..2040
                     /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(1441..1460)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1937..1957
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2702..2719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2812..3400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2892..2911)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3571..4362,4363..4431))
                     /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     4194..4213
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4432..4536)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tccctgtctt aggtcagtta ggatcaccac tttattttaa gaatgtgaaa tatcagaata
       61 atagtagaga gaatgattca tttcagcttt tatttctttc atcacattcc cagtgggtca
      121 gaagtttaca tacactcgac tcctctgcag aatgcggcga tgtttcggta aggggtcctt
      181 aattaatacc acatttgtag aggttttact tgctttaaaa aactctagag aagagcatgc
      241 gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
      301 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga
      361 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
      421 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
      481 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
      541 aattacttcc acgcccctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa
      601 gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg
      661 aggcctggct tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc
      721 tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt
      781 ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt
      841 ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg
      901 gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc
      961 tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt
     1021 cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa
     1081 aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg
     1141 cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc
     1201 acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt
     1261 atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca gcttggcact
     1321 tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc
     1381 ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgaaaa ctacgccacc
     1441 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     1501 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     1561 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     1621 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     1681 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     1741 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     1801 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     1861 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     1921 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     1981 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     2041 taacaactcg agtaccggct taagtgatag atcggaagag cggttcagca ggaatgccga
     2101 gaccggttta aactcagtgt ccatagcgac gccggcgcga taagctggac cccttaccga
     2161 aacatcgccg cattctgcag aggagtcgaa cttagtgtat gtaaacttct gacccactgg
     2221 aattgtgata cagtgaatta taagtgaaat aatctgtctg taaacaattg ttggaaaaat
     2281 gacttgtgtc atgcacaaag tagatgtcct aactgacttg ccaaaactat tgtttgttaa
     2341 caagaaattt gtggagtagt tgaaaaacga gttttaatga ctccaactta agtgtatgta
     2401 aacttccgac ttcaactgta tagagctcct gagatgcagg tacatcccac tgatgagtcc
     2461 caaataggac gaaacgcgct tcggtgcgtc tgggattcca ctgctatcca cgcggccgca
     2521 cggcattacg ttagacgtct aggccacacc accctgaagg cgcctgctcg cctctgatct
     2581 gttgaagcta agcagggtcg gtcctggtta gtacttggat gggactccgc ctggtaatag
     2641 ccggtaccgt aggcttgcat ggtcatagct gtttccttgc gtattgggcg ctctccgctt
     2701 cctcgctcac tgactcgctg cgctcggtcg ttcgggtaaa gcctggggtg cctaatgagc
     2761 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2821 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2881 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2941 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3001 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3061 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3121 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3181 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3241 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3301 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3361 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3421 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3481 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3541 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3601 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3661 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg
     3721 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3781 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3841 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3901 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3961 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4021 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4081 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4141 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4201 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4261 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4321 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4381 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4441 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4501 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4561 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     4621 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     4681 atagggttga gtggccgcta cagggcgctc ccattcgcca ttcaggctgc gcaactgttg
     4741 ggaagggcgt ttcggtgcgg gcctcttcgc tattacgcca gctgggcctt ttgctggcct
     4801 tttgctctat acagttgaag tcggaagttt acatacacct tagccaaata catttaaact
     4861 cactttttca caattcctga catttaatcc tagtaaaaat
//
