LOCUS       sequence-443298-        4925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-12_Xt Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4925)
  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 4925)
  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-Tc1-12_Xt
FEATURES             Location/Qualifiers
     source          1..4925
                     /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        600..1099
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(635..658)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1032..1051
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1113..1122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1119..1718
                     /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     1618..1638
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1725..1781
                     /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(1782..1784,1785..1787,1788..2501)
                     /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(1782..1784,1785..1787,1788..2498)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1827..1848)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2088..2107)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2435..2456
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2523..2540)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2529..2753
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3028..3044)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3028..3044)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3056..3073)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3227..3815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3307..3326)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3986..4777,4778..4846))
                     /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     4609..4628
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4847..4925,1..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 tctgcagtac agtgctttgc taaagtattc accccccttg gcatttttca tgttttgtta
      241 cattccaacc tgtaatttaa atgtttctta atcttatttt atgtgatgga tctgcacaaa
      301 atagtctaag ttggtgaagt gaaatgagaa aaatatatat aaaaaagaat ttaaaaaaaa
      361 taaaaaactg aaaattggca tgtgcatatg tattcacccc ctttgccatg aagcccctaa
      421 aaagtcctgg tgcaaccaat taccttcaaa agtcacataa ttagtgaaat gaagtccacc
      481 tgtgtgcaat ctaagtgtca catgatctgt cagtataaac acaccctttc tgaaaggccc
      541 cagaggcttc aacaccacta agcaagaggc atcacaccac cggtgctagc aattctaccg
      601 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg
      661 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg
      721 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag
      781 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg
      841 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg
      901 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg
      961 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
     1021 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     1081 ctcatctccg ggcctttcga cctgcagcca acgccaccat ggggaccgag tacaagccca
     1141 cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt
     1201 tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca
     1261 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     1321 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     1381 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     1441 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     1501 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     1561 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     1621 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     1681 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccat cgatgccact aacttctccc
     1741 tgttgaaaca agcaggggat gtcgaagaga atcccgggcc aatggtgagc aagggcgagg
     1801 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1861 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1921 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1981 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2041 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2101 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2161 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2221 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2281 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2341 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     2401 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2461 ccgccgggat cactctcggc atggacgagc tgtacaagta attaattaag ctcgctgatc
     2521 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2581 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2641 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2701 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2761 ggcgcgccct cgagctttag ggggggtgaa cagttatgca cgctgaagtt ttctgttatt
     2821 ttgtcctatt tgttgtttgc ttcacaataa aaaaaaatac atcttcaaag ttgtaggcat
     2881 gttctgtaaa tgaaatggtg caaactctca aaacaatcca ttttaattcc aggttgtgag
     2941 gcaacaaaac atgaaaaatg ccaagggggt gaatacttta gcaaagcact gtagggccca
     3001 gagacggagt cactgccaac cgagacggtc atagctgttt cctgtgtgcc gcttcctcgc
     3061 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3121 cggtaatacg gttacccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3181 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3241 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3301 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3361 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3421 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3481 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3541 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3601 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3661 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3721 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3781 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3841 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3901 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3961 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4021 cgatctgtct ctttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4081 tacgggaggg cttaccatct ggccccagtg ctgcaataat accgcgggac ccacgctcac
     4141 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4201 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4261 gttcgccagt taatagtttg cgcaacgttg ttgccatcgc tacaggcatc gtggtatcac
     4321 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4381 gatcccccat gttgcgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4441 gtaagttggc cgccgtgtta tcactcatgg ttatggcagc actacataat tctcttactg
     4501 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4561 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4621 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4681 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4741 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4801 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4861 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4921 tttag
//
