LOCUS       sequence-442197-        5389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of OposCharlie2 Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5389)
  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 5389)
  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-OposCharlie2
FEATURES             Location/Qualifiers
     source          1..5389
                     /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        960..1459
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(995..1018)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1392..1411
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1473..1482
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1479..2078
                     /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     1978..1998
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2085..2141
                     /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(2142..2144,2145..2147,2148..2861)
                     /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(2142..2144,2145..2147,2148..2858)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2187..2208)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2448..2467)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2795..2816
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2883..2900)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2889..3113
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3492..3508)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3492..3508)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3520..3537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3691..4279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3771..3790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4450..5241,5242..5310))
                     /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     5073..5092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5311..5389,1..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 tcaattgctc tagagccgtg atggcgaacc tatgacacgc gtgtcagcac tgacacgcgt
      241 agccattttc gatgacacgc ggcccacata cagagaagta tggggccgca tgccgagaag
      301 gacgtttcat cctcggctcc tgcacggcca ggtgcagtag ccgaggataa aacatttgct
      361 gtagtgtagt gtagacactc tgtgccggag gtctgtaggc caaaacaaca gaactccggc
      421 acagagcgtc tagttctggg acttccggtt aggccgttag gggatctttg acctcacttc
      481 cggccggcgg agcagggagc agcggaatgc cgcgggggac gcatctctgg gggccctgtg
      541 atcgccatta ccagcaacca cataaccaca gcaaccatca tccaatctac tgttctgggg
      601 tgtcgtggat ttctaattga ccatcattac tgagataagt gagggggagg ctgggagagg
      661 cgagggcctg tgctatgggt gccatttccc accattagaa atagcggcag ccatcttaat
      721 ttacataagg aacgccatct tgcagcatag tgcagactcc atttcaccac tattactgtt
      781 gtgcatcctt attaacccct atttctgctt attaaccctg ccctttccta aaagacagtt
      841 acatgtgcca tcttgtggtc agttaggcta gttaacccct aattgcctgc agggctaaca
      901 ggctatctat aattctatct tctgtcactg cccccctgac cggtgctagc aattctaccg
      961 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg
     1021 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg
     1081 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag
     1141 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg
     1201 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg
     1261 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg
     1321 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
     1381 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     1441 ctcatctccg ggcctttcga cctgcagcca acgccaccat ggggaccgag tacaagccca
     1501 cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt
     1561 tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca
     1621 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     1681 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     1741 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     1801 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     1861 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     1921 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     1981 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     2041 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccat cgatgccact aacttctccc
     2101 tgttgaaaca agcaggggat gtcgaagaga atcccgggcc aatggtgagc aagggcgagg
     2161 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2221 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     2281 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2341 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2401 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2461 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2521 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2581 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2641 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2701 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     2761 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2821 ccgccgggat cactctcggc atggacgagc tgtacaagta attaattaag ctcgctgatc
     2881 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2941 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3001 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3061 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3121 ggcgcgccct cgagtaggca ggttagttaa agaattcccc ctccccctca cttatcttag
     3181 ttcacggcac cccacacaag ttaaataata tcaagaccaa caaaagaaac cgactgacag
     3241 atgaacaaag aagtcactaa gcaggtaagt taaataatta gtttttggtt tattaaatac
     3301 agttatatat tacaattata catttttgtt atttaaacta taaatatcgc gaaattatgg
     3361 tttttttctc gaagtgacac accacccgag ttatgctcgg ttttttggcg aattttgaca
     3421 caccaagctc aaaaggttgc ccatcactgc tctagagggg cccagagacg gagtcactgc
     3481 caaccgagac ggtcatagct gtttcctgtg tgccgcttcc tcgctcactg actcgctgcg
     3541 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttacc
     3601 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3661 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3721 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3781 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3841 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3901 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3961 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4021 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4081 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4141 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4201 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4261 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4321 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4381 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4441 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctctttcg
     4501 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4561 atctggcccc agtgctgcaa taataccgcg ggacccacgc tcaccggctc cagatttatc
     4621 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4681 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4741 tttgcgcaac gttgttgcca tcgctacagg catcgtggta tcacgctcgt cgtttggtat
     4801 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgcg
     4861 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgccgt
     4921 gttatcactc atggttatgg cagcactaca taattctctt actgtcatgc catccgtaag
     4981 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5041 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5101 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5161 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5221 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5281 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5341 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttag
//
