LOCUS       sequence-443565-        5388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Zaphod2 Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5388)
  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 5388)
  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-Zaphod2
FEATURES             Location/Qualifiers
     source          1..5388
                     /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        1038..1537
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1073..1096)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1470..1489
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1551..1560
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1557..2156
                     /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     2056..2076
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2163..2219
                     /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(2220..2222,2223..2225,2226..2939)
                     /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(2220..2222,2223..2225,2226..2936)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2265..2286)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2526..2545)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2873..2894
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2961..2978)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2967..3191
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3491..3507)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3491..3507)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3519..3536)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3690..4278)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3770..3789)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4449..5240,5241..5309))
                     /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     5072..5091
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5310..5388,1..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 tcaattgcta tatagcaggg ccgccgcgta cggttgttca ggttgtgcac tgcacaaggg
      241 cgccacatct aagggggcgc cattcacatc gtagacatct ttattatttt gaaacatttt
      301 tatattttta ttataacttt gttgactcca aagatgggcg atttgtatat ttattatgac
      361 aattttccgg cagatggcag taaagtgtct tgttctaaca aaatcagtat attacgacaa
      421 ttttccgaca gatggaagta aagtgtcttg ttctaacagg gttgatatat atcgttccgc
      481 ttcgacttac gaaggaggaa cgagttcgat aatcgaaaga ccttgttgat ctaacaacat
      541 ctaaaataac ctgtgcttct aaacaaggag ctgcttttcc gactgatgta ctgatcggtg
      601 agtatcttaa aatttaaaaa aaaaatcccc tattggtatt agtgcactgg ctaaatttga
      661 tatttcatat tgcagttcat atcatctgcc gtttgtttac tgtaccagaa tggtattgct
      721 gagttcccaa tttaaatcta caagtacttt cattatgaca cgaatcacta tgtaataact
      781 aatattcatt taccattaat aatccacaaa gtaagtatta catagacata aattagagat
      841 tactatgtaa tgatccgcta attatattat agcaattgtt tatttgttgt tatggaaatt
      901 tcgtcagtac aaaaattaac tgctgttaaa atttagcggc ttgttacata catattattt
      961 taaaatgtta caaattctaa gataaaatac taagctatta tattatttct ccccagaccg
     1021 gtgctagcaa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc
     1081 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc
     1141 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc
     1201 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg
     1261 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg
     1321 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc
     1381 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc
     1441 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc
     1501 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagccaac gccaccatgg
     1561 ggaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
     1621 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
     1681 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     1741 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     1801 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     1861 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     1921 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     1981 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     2041 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     2101 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgccatcg
     2161 atgccactaa cttctccctg ttgaaacaag caggggatgt cgaagagaat cccgggccaa
     2221 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2281 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2341 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2401 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2461 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2521 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2581 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2641 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     2701 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2761 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2821 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2881 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
     2941 taattaagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3001 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3061 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3121 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3181 gggctctatg gcttctgagg cgcgccctcg agcaaattat tatatcctag tgactttgag
     3241 ttcctaatta ttgtgattaa aatttcgaat tttctattat tttatgttca ataatttgat
     3301 ttgtataatt tatacagaat gactttatat ttgttgtaat tattattatt gagaaataaa
     3361 taatatacta actttaaaat tttacttgag gaaggggcgc ctttttctaa ttcgcacaaa
     3421 ggcgccgtat gggctagcag cggccctgct atataggggc ccagagacgg agtcactgcc
     3481 aaccgagacg gtcatagctg tttcctgtgt gccgcttcct cgctcactga ctcgctgcgc
     3541 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttaccc
     3601 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3661 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3721 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3781 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3841 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3901 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3961 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4021 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4081 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4141 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4201 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4261 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4321 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4381 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4441 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctctttcgt
     4501 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4561 tctggcccca gtgctgcaat aataccgcgg gacccacgct caccggctcc agatttatca
     4621 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4681 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4741 ttgcgcaacg ttgttgccat cgctacaggc atcgtggtat cacgctcgtc gtttggtatg
     4801 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgcgc
     4861 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgccgtg
     4921 ttatcactca tggttatggc agcactacat aattctctta ctgtcatgcc atccgtaaga
     4981 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5041 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5101 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5161 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5221 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5281 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5341 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttag
//
