LOCUS       sequence-443660-        6243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of PROTOP Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6243)
  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 6243)
  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-PROTOP
FEATURES             Location/Qualifiers
     source          1..6243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..20
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     29..46
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     30..46
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1317..1816
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1352..1375)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1749..1768
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1830..1839
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1836..2435
                     /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     2335..2355
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2442..2498
                     /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(2499..2501,2502..2504,2505..3218)
                     /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(2499..2501,2502..2504,2505..3215)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2544..2565)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2805..2824)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3152..3173
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3240..3257)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3246..3470
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4225..4241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4225..4241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4253..4270)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      4322..4353
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        4354..4456
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4457..4525,4526..5317)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAELGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4675..4694)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             5318..5509
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      5533..5560
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(5572..6159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5652..5671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6181..6210)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 caataggcgt atcacgaggc cctttcgttg taaaacgacg gccagtcgaa ccacgcaatg
       61 cgtctcgatc cgcagtgtct tgcgtctctc atatggtcct ggcaaagaca ctagaataac
      121 aagatgcgta acgccatacg attttttggc acacgatttt ttcgccgtgg ctctagaggt
      181 ggctccaggc tctctcgaat ttttgttcga gagcgagaga gcgaagagcg ctacagcgaa
      241 cagctctttt ctacgcatac agtgatagca gacaactgta tgtgtgcaca cgtatgctca
      301 tgcattgtaa atttgacaaa atatgccctt caccttagaa gttcttagac tttaaatcta
      361 tattattttt gatcaattgg caccatgcga aaaattcttg ttttgcattg ccttaacgtt
      421 attattattt gaaaatagat tagaaatagc caaatctatg tacatattat cacaaaataa
      481 atttcaaaaa tgactttata ttagaatatt tgtcattaga gtattcagct tgcggcgtgt
      541 gaaaaattaa taaggcaatg attgttgagt gcttgtgtcc gcacttcgtg cctcaagata
      601 tgaccaaagc aaagacacta gaataattct agtgtctttg atatgacttt tgcaataaac
      661 agttttcata tttttattta ttttataaat ttttattttc tacttcgtat tatttttatg
      721 aaatatttat ttctcaatgt aatgtcttct ttttgtaaaa tttatgtttt aaattacagt
      781 agttataata atttatattt tacttccttt aattatttag tatatttatt aagtcatttg
      841 acttaatatg atttatgtat gtaaatgatc catttttatt atttaaaatg cacattagat
      901 tcagttgttt ttcagtttgt tttttttttt tgttatattc caattcgata tttttaaaaa
      961 gcgcgccatt tcgggtggga agaaaagcgc gccaattttt ttgtttttcc ttttaatttc
     1021 tcttagcccg tttgcttaaa gttggctata cgcagagctc ctcgattttt tgatattttc
     1081 gctatataat cgaatttata ttcaactaaa gtcgttttgt tgcaaaagta aagtacggaa
     1141 atataagcgc aaatagtaat tgcgataaat agtgaatagt gaatagtgtt tacgtatttt
     1201 cgtaatttga cgcgctaata caacgtgctg ctgcttttcg acaaaggttt gagttgaaat
     1261 ttataaattg taagtgaaaa aaaaaaaaca ttttaaccgg tgctagcaat tctaccgggt
     1321 aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac
     1381 ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca
     1441 accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa
     1501 gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct
     1561 cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg
     1621 cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg
     1681 gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg
     1741 aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc
     1801 atctccgggc ctttcgacct gcagccaacg ccaccatggg gaccgagtac aagcccacgg
     1861 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg
     1921 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg
     1981 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     2041 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     2101 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     2161 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     2221 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     2281 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     2341 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     2401 cgcgcacctg gtgcatgacc cgcaagcccg gtgccatcga tgccactaac ttctccctgt
     2461 tgaaacaagc aggggatgtc gaagagaatc ccgggccaat ggtgagcaag ggcgaggagc
     2521 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2581 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     2641 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     2701 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2761 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2821 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2881 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2941 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     3001 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     3061 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     3121 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     3181 ccgggatcac tctcggcatg gacgagctgt acaagtaatt aattaagctc gctgatcagc
     3241 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3301 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3361 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3421 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3481 gcgccctcga gaaaaaatga gcgatattta aataactgac gactgaaaaa tcgagcaata
     3541 tataataaaa aacaatagtt agaaaaataa cttaattggg gaacatggaa atgttttaat
     3601 gttaaacatt aaaatatttc aagtcgactc gaaggtcata tacgtaaata taggacccca
     3661 ttacaattgt aatggcctcc ccgtggtgtt ccctgggtac cgattattac acatattaca
     3721 cataattaat tatcaacata aatataaata tgtaaacggt agctaattcg agcggcgatt
     3781 ttaacaaacg aatttaaaaa gctttaaaat taaaaaattt taaaattaaa attaaaattt
     3841 taaaattata ataatcaggg cgcgaatttt taaaaattat tttatttcat catattgcta
     3901 cgaaattggc aaaaaactac cctaatatgt acaatgtaaa ttcatttctt cgatcagaat
     3961 tgatttcggc ccgaaaatcg tcttctagca caacacgcac acatatacgc gttctcgtct
     4021 cttgttttta ctcacacaag caagcaaatt ctatttttag atttcttacg ttctcagcgt
     4081 gagcgagcgg aaagagagca attttggccg tcaccaaaaa agtggctgca tagtgccaaa
     4141 ccaatgtatg gccgttacgc atcttgttat tctagtgtct ttggtcctgg gggcccagag
     4201 acggagtcac tgccaaccga gacggtcata gctgtttcct gtgtgccgct tcctcgctca
     4261 ctgactcgct gctctcggtc gttactgacc atttaaatca tacctgacct ccatagcaga
     4321 aagtcaaaag cctccgaccg gaggcttttg acttgatcgg cacgtaagag gttccaactt
     4381 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     4441 agctaaggaa gctaaaatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4501 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4561 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4621 tgagagttta cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4681 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4741 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctca cggatggcat
     4801 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4861 acttctggca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4921 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4981 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5041 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5101 aggatcactt ctgcgctcgg ccctcccggc tggctggttt attgctgata aatctggagc
     5161 cggtgagcgt gggtcccgcg gtatcattgc agaactgggg ccagatggta agccctcccg
     5221 catcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5281 cgctgagata ggtgcctcac tgattaagca ttggtaagtg accaaacagg aaaaaaccgc
     5341 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
     5401 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
     5461 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgat gagggcccaa
     5521 atgtaatcac ctggctcacc ttcgggtggg cctttctgcg ttgctggcgt ttttccatag
     5581 gctccgcccc cctgacgagc atcacaaaaa tcgatgctca agtcagaggt ggcgaaaccc
     5641 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5701 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5761 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5821 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5881 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5941 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6001 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta cctcggaaaa
     6061 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6121 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gattttctac
     6181 cgaagaaagg cccacccgtg aaggtgagcc agtgagttga ttgcagtcca gttacgctgg
     6241 agt
//
