LOCUS       sequence-441627-        5666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of piggyBac-1_AMi Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5666)
  AUTHORS   Zhang T, Tan S, Tang N, Li Y, Zhang C, Sun J, Guo Y, Gao H, Cai Y, 
            Sun W, Wang C, Fu L, Ma H, Wu Y, Hu X, Zhang X, Gee P, Yan W, Zhao 
            Y, Chen Q, Guo B, Wang H, Zhang YE
  TITLE     Heterologous survey of 130 DNA transposons in human cells highlights
            their functional divergence and expands the genome engineering 
            toolbox.
  JOURNAL   Cell. 2024 Jul 11;187(14):3741-3760.e30. doi: 
            10.1016/j.cell.2024.05.007. Epub 2024 Jun 5.
  PUBMED    38843831
REFERENCE   2  (bases 1 to 5666)
  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.2024.05.007"; journalName: "Cell"; date: 
            "2024-07-11- 11"; volume: "187"; issue: "14"; pages: "3741-3760.e30"
COMMENT     Sequence Label: pMV-piggyBac-1_AMi
FEATURES             Location/Qualifiers
     source          1..5666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..112
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     121..138
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     122..138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1205..1704
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1240..1263)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1637..1656
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1718..1727
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1724..2323
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     2223..2243
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2330..2386
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(2387..2389,2390..2392,2393..3106)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2387..2389,2390..2392,2393..3103)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2432..2453)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2693..2712)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3040..3061
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3128..3145)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3134..3358
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3769..3785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3769..3785)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3797..3814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3968..4556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4048..4067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4727..5518,5519..5587))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLRNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERKRQIAEIGAS
                     LIKHW"
     primer_bind     5350..5369
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5588..5666,1..26))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
       61 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
      121 tgtaaaacga cggccagtcg aaccacgcaa tgcgtctcga tccgcagtgt cttgcgtctc
      181 tcaattgtta accttcatac gttcccatgg ggtaatttcc taccccagac gcatattttg
      241 tgcagtatct taaagttaat taattctaaa aaatattctt tttaggacat tgtcactggc
      301 accgcagtgc acaatatccc caaagaacag ccagcttaca tgaatggttt gctctctgtg
      361 gatgaaaaac tgcccagggt aacatgttac cccaggcgtg aatgtgtttt tccatatggg
      421 gtaacaactt accccaagcc catgttaacg cattggggta acttgttacc ccaatgcaat
      481 ttctgtataa atgacaggcc tagacctgtt tggccacttt gggtggatgc atctgaaagg
      541 tcagcattca ttcacacctc attcatgtcc acttcaggta tccatggtaa ctaatggatg
      601 gccatcctcc actctgcttt tgcagaagat gtggtaatcc cttttgctaa ccactttgtt
      661 tgcaggctgc aaggtcatca aaacaccctg ccctttgcca aaaaaaccca aaaccctttg
      721 tcacacccaa atccacctct gctctctttt ctcatggcaa aactccgcct cacctgcccc
      781 tttcataaca tacaagctag tttgccctgc ctttgttgga agtgaggaca aggagcaaag
      841 ggacagcaga gaagaaatag aacaacatca tactgcaata cagcgtacac tatggagtcc
      901 ccttcaacgg taagggcttg acacccccca tgcccctttt acctgctctc aataagcccc
      961 tgcatgtgtg gtttccaaac agcatccacc cacatttggc ttgcctgcaa agggtggatg
     1021 ctggtctctg gcttctgtgc ccaggaggag agggtgctac ctgcgctgcc atagggcagg
     1081 gcggaattgc ttctgtgaca gcataggtgg acaggcttgt ctttgattgt tatttatatg
     1141 cattcagcag taatcatttt tttttttttt tctgcagtcc aggaccggtg ctagcaattc
     1201 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     1261 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     1321 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     1381 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     1441 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     1501 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     1561 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg
     1621 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     1681 tcttcctcat ctccgggcct ttcgacctgc agccaacgcc accatgggga ccgagtacaa
     1741 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc
     1801 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg
     1861 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     1921 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     1981 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     2041 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     2101 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     2161 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     2221 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     2281 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gccatcgatg ccactaactt
     2341 ctccctgttg aaacaagcag gggatgtcga agagaatccc gggccaatgg tgagcaaggg
     2401 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2461 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2521 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2581 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2641 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2701 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2761 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2821 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     2881 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     2941 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     3001 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3061 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaattaa ttaagctcgc
     3121 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3181 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3241 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3301 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3361 tctgaggcgc gccctcgagt taaaacagtt aggaaatgtt tttggaagta attgttagtt
     3421 tgaaaacagt gtgaactgca gacttagcaa attgttcatt cacgttttga aactctaaaa
     3481 aaagtttaaa ttttgttcat agttctagtt aaataatatt ttgggttcaa atcatcataa
     3541 atactctttc aaacaatctg tatgtttaat aaatagtttt tggtacattt atactgttgt
     3601 cactggttcc attttgttca tgcgggtggg gtaacaagtt accccatgag accaacaatg
     3661 tcgaattgcg gggtaacaag ttaccccatg agaacaacaa cggttacggt ttcatgagaa
     3721 cggatgaggg ttaagggccc agagacggag tcactgccaa ccgagacggt catagctgtt
     3781 tcctgtgtgc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3841 cggtatcagc tcactcaaag gcggtaatac ggttacccac agaatcaggg gataacgcag
     3901 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3961 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4021 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4081 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4141 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4201 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4261 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4321 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4381 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4441 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4501 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4561 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4621 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4681 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4741 tcagtgaggc acctatctca gcgatctgtc tctttcgttc atccatagtt gcctgactcc
     4801 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaataa
     4861 taccgcggga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4921 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4981 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccatcg
     5041 ctacaggcat cgtggtatca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5101 aacgatcaag gcgagttaca tgatccccca tgttgcgcaa aaaagcggtt agctccttcg
     5161 gtcctccgat cgttgtcaga agtaagttgg ccgccgtgtt atcactcatg gttatggcag
     5221 cactacataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5281 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5341 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5401 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5461 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5521 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5581 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5641 gcggatacat atttgaatgt atttag
//
