LOCUS       sequence-443292-        4836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-5_Xt Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4836)
  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 4836)
  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-Tc1-5_Xt
FEATURES             Location/Qualifiers
     source          1..4836
                     /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        508..1007
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(543..566)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     940..959
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1021..1030
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1027..1626
                     /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     1526..1546
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1633..1689
                     /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(1690..1692,1693..1695,1696..2409)
                     /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(1690..1692,1693..1695,1696..2406)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1735..1756)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1996..2015)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2343..2364
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2431..2448)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2437..2661
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2939..2955)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2939..2955)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2967..2984)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3138..3726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3218..3237)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3897..4688,4689..4757))
                     /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     4520..4539
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4758..4836,1..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 aaaccggatt ccaaaaaagt tgggacacta aacaaattgt gaataaaaac
      241 tgaacgcaat gatgtggagg tgccaacttc taatatttta ttcagaatag aacataaatc
      301 acggaacaaa agtttaaact gagaaaatgt accattttaa gggaaaaata tgttgattca
      361 gaatttcatg gtgtcaacaa atcccaaaaa agttgggaca agtagcaata agaggctgga
      421 aaaagtaaat ttgagcataa cgaagagctg gaagaccaaa taacactaat taggtcaatt
      481 ggcaacaccg gtgctagcaa ttctaccggg taggggaggc gcttttccca aggcagtctg
      541 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc
      601 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
      661 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt
      721 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg
      781 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg
      841 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca
      901 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc
      961 gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagccaac
     1021 gccaccatgg ggaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     1081 cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     1141 gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     1201 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     1261 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     1321 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     1381 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     1441 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     1501 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     1561 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     1621 ggtgccatcg atgccactaa cttctccctg ttgaaacaag caggggatgt cgaagagaat
     1681 cccgggccaa tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     1741 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1801 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1861 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     1921 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1981 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2041 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2101 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2161 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     2221 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2281 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     2341 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2401 tacaagtaat taattaagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     2461 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2521 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2581 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2641 gggatgcggt gggctctatg gcttctgagg cgcgccctcg agaaatggcc ttgtcccaac
     2701 ttttttggga tttgttgaca ccatgaaatt ctgaatcaac atatttttcc cttaaaatgg
     2761 tacattttct cagtttaaac ttttgttccg tgatttatgt tctattctga ataaaatatt
     2821 agaagttggc acctccacat cattgcgttc agtttttatt cacaatttgt ttagtgtccc
     2881 aacttttttg gaatccgggt tgtagggccc agagacggag tcactgccaa ccgagacggt
     2941 catagctgtt tcctgtgtgc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3001 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttacccac agaatcaggg
     3061 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3121 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3181 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3241 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3301 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3361 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3421 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3481 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3541 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3601 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3661 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3721 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3781 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3841 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3901 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tctttcgttc atccatagtt
     3961 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4021 gctgcaataa taccgcggga cccacgctca ccggctccag atttatcagc aataaaccag
     4081 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4141 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4201 gttgccatcg ctacaggcat cgtggtatca cgctcgtcgt ttggtatggc ttcattcagc
     4261 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgcgcaa aaaagcggtt
     4321 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgccgtgtt atcactcatg
     4381 gttatggcag cactacataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4441 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4501 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4561 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4621 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4681 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4741 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4801 tgtctcatga gcggatacat atttgaatgt atttag
//
