LOCUS       sequence-442185-        4935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-9_Xt Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4935)
  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 4935)
  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-Tc1-9_Xt
FEATURES             Location/Qualifiers
     source          1..4935
                     /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        572..1071
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(607..630)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     regulatory      1085..1094
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1091..1690
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTGRLATRDDVPRAARTLAAALADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     1590..1610
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1697..1753
                     /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(1754..1756,1757..1759,1760..2473)
                     /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(1754..1756,1757..1759,1760..2470)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1799..1820)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2060..2079)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2407..2428
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2495..2512)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2501..2725
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3038..3054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3038..3054)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3066..3083)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3237..3825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3317..3336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3996..4787,4788..4856))
                     /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     4619..4638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4857..4935,1..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 agttaggtcc ataaatattt ggacagagac aacttttttc taattttggt
      241 tctgtacatt accacaatga attttaaatg aaacaactca gatgcagttg aagtgcagac
      301 tttcagcttt aattcagtgg ggtgaacaaa acgattgcat aaaaatgtga ggccactaaa
      361 gcattttttt taacacaatc ccttcatttc aagggctcaa aagtaattgg acaattgact
      421 caaaggctat ttcatgggca ggtgttggca agtccgtcgt tatgtcatta tcaattaagc
      481 agataaaagg cctggagttg atttgaggtg tggtgcttgc atgtggaaga ttttgctgtg
      541 aacagacaac accggtgcta gcaattctac cgggtagggg aggcgctttt cccaaggcag
      601 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
      661 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
      721 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
      781 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc
      841 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
      901 ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg
      961 ctcaggggcg gggcgggcgc ccgaagggcc tccgggggcc cggcattctg cacgcttcaa
     1021 aaacgcacgt ctgccgcgct gttctcctcc tcctcatctc cgggcctttc gacccgcagc
     1081 caacgccacc atggggaccg agtacaagcc cacggggcgc ctcgccaccc gcgacgacgt
     1141 cccccgggcc gcacgcaccc tcgccgccgc gctcgccgac taccccgcca cgcgccacac
     1201 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     1261 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     1321 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     1381 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     1441 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     1501 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     1561 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     1621 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     1681 gcccggtgcc atcgatgcca ctaacttctc cctgttgaaa caagcagggg atgtcgaaga
     1741 gaatcccggg ccaatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1801 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1861 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1921 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1981 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2041 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2101 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2161 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2221 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2281 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2341 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2401 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2461 gctgtacaag taattaatta agctcgctga tcagcctcga ctgtgccttc tagttgccag
     2521 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2581 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2641 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2701 gctggggatg cggtgggctc tatggcttct gaggcgcgcc ctcgagaaat gaacatttta
     2761 tttccagtta tttaatttgt ccaattactt ttgagcccct gaaatgaagg gattgtgtta
     2821 aaaaaaatgc tttagtggcc tcacattttt atgcaatcgt tttgttcacc ccactgaatt
     2881 aaagctgaaa gtctgcactt caactgcatc tgagttgttt catttaaaat tcattgtggt
     2941 aatgtacaga accaaaatta gaaaaaagtt gtctctgtcc aaatatttat ggacctaact
     3001 gtagggccca gagacggagt cactgccaac cgagacggtc atagctgttt cctgtgtgcc
     3061 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3121 cactcaaagg cggtaatacg gttacccaca gaatcagggg ataacgcagg aaagaacatg
     3181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3661 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4021 cctatctcag cgatctgtct ctttcgttca tccatagttg cctgactccc cgtcgtgtag
     4081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaataat accgcgggac
     4141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4201 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccatcgc tacaggcatc
     4321 gtggtatcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4381 cgagttacat gatcccccat gttgcgcaaa aaagcggtta gctccttcgg tcctccgatc
     4441 gttgtcagaa gtaagttggc cgccgtgtta tcactcatgg ttatggcagc actacataat
     4501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4621 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4861 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4921 tttgaatgta tttag
//
