LOCUS       sequence-443303-        5313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Chaplin1_DR Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5313)
  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 5313)
  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-Chaplin1_DR
FEATURES             Location/Qualifiers
     source          1..5313
                     /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        530..1029
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(565..588)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     962..981
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1043..1052
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1049..1648
                     /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     1548..1568
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1655..1711
                     /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(1712..1714,1715..1717,1718..2431)
                     /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(1712..1714,1715..1717,1718..2428)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1757..1778)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2018..2037)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2365..2386
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2453..2470)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2459..2683
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3416..3432)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3416..3432)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3444..3461)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3615..4203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3695..3714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4374..5165,5166..5234))
                     /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     4997..5016
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5235..5313,1..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 tactagtgtc tatagcaggg gtcggcaacc cgcggctcta gagccgcatg cggctcttta
      241 gcgctgccct agtggctccc tggagctttt tcaataaatt ttgaaaatga aaaaagatgt
      301 gggaggtaaa tatagttttt gtttttaata tggtttctat aggaggaaaa gttttccaat
      361 gctgtaaaag tgtgtagaat aaatatttaa tttcaacatt tctgtcaacg aagatttgcg
      421 tcatggtctg cgacacacgt ttccatcagc agggcgggat gccagacagg tggctattgt
      481 aaataaaccg gcggctgtgt gatggacgac cggtgctagc aattctaccg ggtaggggag
      541 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
      601 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
      661 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
      721 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
      781 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
      841 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
      901 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
      961 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     1021 ggcctttcga cctgcagcca acgccaccat ggggaccgag tacaagccca cggtgcgcct
     1081 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     1141 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     1201 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     1261 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     1321 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     1381 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     1441 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     1501 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     1561 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     1621 ctggtgcatg acccgcaagc ccggtgccat cgatgccact aacttctccc tgttgaaaca
     1681 agcaggggat gtcgaagaga atcccgggcc aatggtgagc aagggcgagg agctgttcac
     1741 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1801 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     1861 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     1921 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1981 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2041 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2101 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2161 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     2221 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2281 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     2341 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2401 cactctcggc atggacgagc tgtacaagta attaattaag ctcgctgatc agcctcgact
     2461 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2521 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2581 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2641 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcgcgccct
     2701 cgagaagggt aagaatcctg tcataggcac gtatttagtg gtagttataa agtgatttct
     2761 tatttttgtc agtatatatt tggaatgaca cttaagggat atgattgtgt tttgataggt
     2821 ccgaggtagc tgcgttggtt gcgtgtcaaa agagaagagg atgctggatt tcacccttgc
     2881 actaacttgc ttggcatttg cacagaaatc aaacttaaac tatattaatt tgattttata
     2941 tacttttttt aaaaggctgc tgttttattt ttttataaat gcagtctgcg ttttattgca
     3001 ccgtttgttg ctctgataag gggcacgtta tgcacgttcc attttgatgt tgtttttgaa
     3061 atccttaaaa taaaaatttt atcaaaaatc taatttcttt attacattta ttttatttta
     3121 tcaaagcaat gaaacataat cccttaatat tgtaatgaag ttaaacttga ggctgcatcg
     3181 tacaacagag tagtcacgtg gtgcgtcatt ctacaggatg cactgcaggg aaaataatag
     3241 atacatatgg catttgttgc cttcattata aggcttaaat aaggctttta attttctgcg
     3301 gctccagaaa tatttgtttt ttgttttttt ggtccaatat ggctctttca acattttggg
     3361 ttgccgaccc ctggtctata ggggcccaga gacggagtca ctgccaaccg agacggtcat
     3421 agctgtttcc tgtgtgccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3481 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tacccacaga atcaggggat
     3541 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3601 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3661 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3721 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3781 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3841 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3901 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3961 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4021 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4081 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4141 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4201 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4261 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4321 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4381 tgcttaatca gtgaggcacc tatctcagcg atctgtctct ttcgttcatc catagttgcc
     4441 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4501 gcaataatac cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4561 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4621 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4681 gccatcgcta caggcatcgt ggtatcacgc tcgtcgtttg gtatggcttc attcagctcc
     4741 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgcgcaaaaa agcggttagc
     4801 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg ccgtgttatc actcatggtt
     4861 atggcagcac tacataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4921 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4981 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5041 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5101 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5161 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5221 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5281 ctcatgagcg gatacatatt tgaatgtatt tag
//
