LOCUS       sequence-441873-        4899 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Donor plasmid of Tc1-14_Xt Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4899)
  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 4899)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Tc1-14_Xt
FEATURES             Location/Qualifiers
     source          1..4899
                     /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"
     primer_bind     1004..1023
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward 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="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     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(3002..3018)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3002..3018)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3030..3047)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3201..3789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3281..3300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3960..4751,4752..4820))
                     /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     4583..4602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4821..4899,1..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 tcaattgtac agtggttctc gaaagtttgt gaacccttta aaattttcta tatttttata
      241 tgaatgtgac ctaaaacatc atctgatttt caaacaagtc ctaaaagtag atgaagaaaa
      301 cctagttaaa caaatgaaac aaaaattatt atatttggtc atgtatttat tgaaaaaaaa
      361 atgatccaat aacatatctg cgtgtggcaa aagtaagtga atccttagga ttatcatata
      421 atttgaaggt gaaatcagag tctggtgttt tcagtcaatg ggatgacaat caggtgtgag
      481 tgagagaccc tgttttattt aaagaacggg gatccagcaa agcctgatca cacatacaac
      541 acatttgtgg 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 ccgaaggtcc tccggrggcc cggcattctg cacgcttcaa
     1021 aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc
     1081 caacgccacc atggggaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     1141 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac 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 ctcgaggagc acgattcact
     2761 tacttttgcc acacgcagat atgttattgg atcatttttt ttcaataaat acatgaccaa
     2821 atataataat ttttgtttca tttgtttaac taggttttct tcatctactt ttaggacttg
     2881 tttgaaaatc agatgatgtt ttaggtcaca ttcatataaa aatatagaaa attttaaagg
     2941 gttcacaaac tttcaagcac cactgtaggg cccagagacg gagtcactgc caaccgagac
     3001 ggtcatagct gtttcctgtg tgccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3061 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttacc cacagaatca
     3121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3601 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3901 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3961 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctctttcg ttcatccata
     4021 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4081 agtgctgcaa taataccgcg ggacccacgc tcaccggctc cagatttatc agcaataaac
     4141 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4201 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4261 gttgttgcca tcgctacagg catcgtggta tcacgctcgt cgtttggtat ggcttcattc
     4321 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgcg caaaaaagcg
     4381 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgccgt gttatcactc
     4441 atggttatgg cagcactaca taattctctt actgtcatgc catccgtaag atgcttttct
     4501 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4561 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4621 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4681 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4741 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4801 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4861 tattgtctca tgagcggata catatttgaa tgtatttag
//
