LOCUS       sequence-442203-        4892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-4_Xt Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4892)
  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 4892)
  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-Tc1-4_Xt
FEATURES             Location/Qualifiers
     source          1..4892
                     /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        568..1067
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(603..626)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1000..1019
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1081..1090
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1087..1686
                     /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     1586..1606
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1693..1749
                     /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(1750..1752,1753..1755,1756..2469)
                     /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(1750..1752,1753..1755,1756..2466)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1795..1816)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2056..2075)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2403..2424
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2491..2508)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2497..2721
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2995..3011)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2995..3011)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3023..3040)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3194..3782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3274..3293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3953..4744,4745..4813))
                     /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     4576..4595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4814..4892,1..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 taagctttac agtgaggaac ataagtattt gaacaccctg cgattttgca agttctccca
      241 cttagaaatc atggaggggt ctgaaattca cattgtaggt gcattcccac tgtgagagac
      301 agcatttaaa aaaaaaattc aggaaatcac attgtatgat ttttaaagaa tgtatttgta
      361 ttgcactgct gcacataagt atttgaacac ctggcaatca gcaagaattc tggctctcaa
      421 agacctgtta ctctgccttt aaaaagtcca cctctactcc actcattaat ctaaattagt
      481 agcacctgtc tgagctcttt aaagacacct gtccacccca cagtcagtca gactccaact
      541 actaccaccg gtgctagcaa ttctaccggg taggggaggc gcttttccca aggcagtctg
      601 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc
      661 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
      721 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt
      781 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg
      841 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg
      901 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca
      961 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc
     1021 gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagccaac
     1081 gccaccatgg ggaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     1141 cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     1201 gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     1261 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     1321 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     1381 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     1441 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     1501 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     1561 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     1621 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     1681 ggtgccatcg atgccactaa cttctccctg ttgaaacaag caggggatgt cgaagagaat
     1741 cccgggccaa tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1801 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1861 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1921 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     1981 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2041 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2101 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2161 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2221 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     2281 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2341 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2401 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2461 tacaagtaat taattaagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     2521 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2581 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2641 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2701 gggatgcggt gggctctatg gcttctgagg cgcgccctcg agcactgatt ttctcaggtg
     2761 ttcaaatact tatgtgcagc agtgcaatac aaatacattc tttaaaaatc atacaatgtg
     2821 atttcctgaa tttttttttt aaatgctgtc tctcacagtg ggaatgcacc tacaatgtga
     2881 atttcagacc cctccatgat ttctaagtgg gagaacttgc aaaatcgcag ggtgttcaaa
     2941 tacttatgtt cctcactgta gggcccagag acggagtcac tgccaaccga gacggtcata
     3001 gctgtttcct gtgtgccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3061 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt acccacagaa tcaggggata
     3121 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3181 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3241 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3301 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3361 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3421 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3481 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3541 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3601 tgaagtggtg gcctaactac ggctacacta gaagaacagt atctggtatc tgcgctctgc
     3661 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3721 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3781 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3841 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3901 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3961 gcttaatcag tgaggcacct atctcagcga tctgtctctt tcgttcatcc atagttgcct
     4021 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4081 caataatacc gcgggaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4141 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4201 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4261 ccatcgctac aggcatcgtg gtatcacgct cgtcgtttgg tatggcttca ttcagctccg
     4321 gttcccaacg atcaaggcga gttacatgat cccccatgtt gcgcaaaaaa gcggttagct
     4381 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc cgtgttatca ctcatggtta
     4441 tggcagcact acataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4501 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4561 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4621 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4681 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4741 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4801 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4861 tcatgagcgg atacatattt gaatgtattt ag
//
