LOCUS       sequence-443563-        5535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Zaphod3 Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5535)
  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 5535)
  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-Zaphod3
FEATURES             Location/Qualifiers
     source          1..5535
                     /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        1093..1592
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1128..1151)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1525..1544
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1606..1615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1612..2211
                     /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     2111..2131
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2218..2274
                     /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(2275..2277,2278..2280,2281..2994)
                     /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(2275..2277,2278..2280,2281..2991)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2320..2341)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2581..2600)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2928..2949
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3016..3033)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3022..3246
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3638..3654)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3638..3654)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3666..3683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3837..4425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3917..3936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4596..5387,5388..5456))
                     /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     5219..5238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5457..5535,1..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 tatagcagtg gcgtaccaag ggcggggcgg tgggagcggt ccgccccggg
      241 tgcaggcaat aagggggtgc attgtctgta gagaatttaa aaacaataat aaaaccgact
      301 aaaagtcggt ctgcttttta ttatcaccat gcgccggcaa ttctaaacaa tgtcagtgat
      361 aaaatactcc tccccgaaaa atcttttgtt ggtctaagtt ctaaacaatt gctgcggtta
      421 ctgttgagtt ttaataatat atatgtaagc ttcaaattag cacattttta ttacttatcc
      481 tttaataaac attgtattct acatggaagt taattcggag aactcccagt tatacagtcg
      541 gcccccgaca cacgcggact cagctacacg cgttcgtttc gagagtaagt tcgtaacggt
      601 tcggaatcgt tcgagctcgc ttcgggcgca gttcgtgtct ccaacccctg tggtactaca
      661 tattcctgcg tttaaacagt agattcgaaa taaacaatga tagcacagtg attgtaaaga
      721 cgaagaaaca gaacttgagt tacttcaatt ctgtcattct atgtgaccac ttggagtttt
      781 tatttgtgtt taaaatttaa aacagtgaaa cagagtgcga actgcgaggt gtaatatttt
      841 tgtttggtaa gtgcaaattt tagttcatac atgaaatatt ttactgaatt tgaataatat
      901 ctttaaaatt gaaatttatt ctttttaaat tgttaattgt tttaaaacta aagaatgaat
      961 caagaaaata aaatattaca tcagtggtac aatttagtag tggcagatat taaaagcaaa
     1021 gtactttgga attctttttg atagcactcc cgatgtttcg cacctgatca aaccggtgct
     1081 agcaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag
     1141 cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat
     1201 ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc
     1261 ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa
     1321 tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc
     1381 gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga
     1441 ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg
     1501 cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc
     1561 tgttctcctc ttcctcatct ccgggccttt cgacctgcag ccaacgccac catggggacc
     1621 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc
     1681 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac
     1741 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     1801 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     1861 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     1921 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     1981 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     2041 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     2101 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     2161 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc catcgatgcc
     2221 actaacttct ccctgttgaa acaagcaggg gatgtcgaag agaatcccgg gccaatggtg
     2281 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2341 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2401 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2461 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2521 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2581 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2641 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2701 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2761 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2821 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2881 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2941 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaattaatt
     3001 aagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3061 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3121 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3181 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3241 ctatggcttc tgaggcgcgc cctcgagtgt tattattcat tactgcgaca gaccaatatg
     3301 taggtataag ttttttcctt ttttcaaaaa atacattaat gtaattaaaa agtattaatc
     3361 cattactttt tttccttttt tgtactgtaa tatttatttt ttatttttta tactggcatg
     3421 attatatata caaagttcaa taaaagaaaa ttttcactgt ctgcgtttct tttctggcca
     3481 ttattattat tcgtttcatt tcatgattat tactgaaaat aattttgtcg tatagaggag
     3541 gggggtgtta aaaaatgatc cgctccgggt gtcaaatacg ctaggtacgc cactgctata
     3601 taggggccca gagacggagt cactgccaac cgagacggtc atagctgttt cctgtgtgcc
     3661 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3721 cactcaaagg cggtaatacg gttacccaca gaatcagggg ataacgcagg aaagaacatg
     3781 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3841 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3901 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3961 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4021 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4081 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4141 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4201 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4261 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4321 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4381 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4441 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4501 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4561 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4621 cctatctcag cgatctgtct ctttcgttca tccatagttg cctgactccc cgtcgtgtag
     4681 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaataat accgcgggac
     4741 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4801 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4861 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccatcgc tacaggcatc
     4921 gtggtatcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4981 cgagttacat gatcccccat gttgcgcaaa aaagcggtta gctccttcgg tcctccgatc
     5041 gttgtcagaa gtaagttggc cgccgtgtta tcactcatgg ttatggcagc actacataat
     5101 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5161 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5221 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5281 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5341 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5401 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5461 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5521 tttgaatgta tttag
//
