LOCUS       sequence-442030-        4895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of TC1_FR2 Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4895)
  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 4895)
  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_FR2
FEATURES             Location/Qualifiers
     source          1..4895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(140..728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(220..239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(899..1690,1691..1759))
                     /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     1522..1541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1760..1864)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1932..1950
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     1959..1976
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1960..1976
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2367..2866
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2402..2425)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2799..2818
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      2880..2889
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2886..3485
                     /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     3385..3405
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3492..3548
                     /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(3549..3551,3552..3554,3555..4268)
                     /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(3549..3551,3552..3554,3555..4265)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3594..3615)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3855..3874)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4202..4223
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4290..4307)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4296..4520
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4836..4852)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4836..4852)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4864..4881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttaccc acagaatcag
       61 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      121 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      181 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      241 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      301 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      361 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      421 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      481 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      541 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      601 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      661 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      721 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      781 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      841 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      901 accaatgctt aatcagtgag gcacctatct cagcgatctg tctctttcgt tcatccatag
      961 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1021 gtgctgcaat aataccgcgg gacccacgct caccggctcc agatttatca gcaataaacc
     1081 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1141 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1201 ttgttgccat cgctacaggc atcgtggtat cacgctcgtc gtttggtatg gcttcattca
     1261 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgcgc aaaaaagcgg
     1321 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgccgtg ttatcactca
     1381 tggttatggc agcactacat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1441 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1501 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1561 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1621 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1681 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1741 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1801 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1861 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1921 taacctataa aaataggcgt atcacgaggc cctttcgttg taaaacgacg gccagtcgaa
     1981 ccacgcaatg cgtctcgatc cgcagtgtct tgcgtctcta agctttacac tgctcaaaaa
     2041 aattagagga acacttttta atcagagtat agcaacctat cagtcaaact tctgggatat
     2101 tgatctggtc agttaagtag cagagggggt tgttaatcag tttctgctgc tttgttgtta
     2161 atgaaatcaa caacaggagc atcagagggg caacaatgag acggccccca aaacaggaat
     2221 ggctttccag gttgaggcca ctgatacttt tttccctcct catctctttt ggctgatttt
     2281 ttactgactg actttctact gctgtagtta ttcatttggc ttggatcaac atctctgttg
     2341 atagcaccgg tgctagcaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     2401 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     2461 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     2521 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
     2581 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
     2641 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
     2701 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
     2761 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
     2821 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagccaacg
     2881 ccaccatggg gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc
     2941 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     3001 acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     3061 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     3121 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     3181 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     3241 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     3301 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     3361 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     3421 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     3481 gtgccatcga tgccactaac ttctccctgt tgaaacaagc aggggatgtc gaagagaatc
     3541 ccgggccaat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     3601 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     3661 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     3721 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     3781 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     3841 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     3901 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     3961 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     4021 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     4081 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     4141 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     4201 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     4261 acaagtaatt aattaagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     4321 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     4381 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4441 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4501 ggatgcggtg ggctctatgg cttctgaggc gcgccctcga ggaatcattt tgagttgcta
     4561 caatgacatt ttggcaaaat ggaccagtct gctgcatcat tttttcactt tcatttttgg
     4621 ggtgtctttg atttcccccc tctatagggt gatcattttc atttctatca aattatgtgg
     4681 catcattttg ttactaatac atcacctact ttttatcagg aaagatattc aagatcattt
     4741 tcccccccgt ttagatctga tgtgttttcg aagtgttcct ctaatttttt tgagcagtgt
     4801 agggcccaga gacggagtca ctgccaaccg agacggtcat agctgtttcc tgtgtgccgc
     4861 ttcctcgctc actgactcgc tgcgctcggt cgttc
//
