LOCUS       sequence-441686-        5908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tigger2f Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5908)
  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 5908)
  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-Tigger2f
FEATURES             Location/Qualifiers
     source          1..5908
                     /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        813..1312
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(848..871)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1245..1264
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1326..1335
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1332..1931
                     /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     1831..1851
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1938..1994
                     /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(1995..1997,1998..2000,2001..2714)
                     /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(1995..1997,1998..2000,2001..2711)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2040..2061)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2301..2320)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2648..2669
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2736..2753)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2742..2966
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4011..4027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4011..4027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4039..4056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4210..4798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4290..4309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4969..5760,5761..5829))
                     /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     5592..5611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5830..5908,1..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 agttgaccct tgaacaacac gggtttgaac tgcgcgggtc cacttacacg
      241 cggatttttt tcaataaata tattggaaaa ttttttggag atttgtgaca atttgaaaaa
      301 actcacagac gaaccgcgta gcctagaaat atcgaaaaaa ttaagaaaaa gttaggtatg
      361 tcatgaatgc ataaaatata tgtagatact agtctatttt atcatttact accataaaat
      421 atacacaaat ctattataaa aagttaaaat ttatcaaaac ttatgcacac acttacagac
      481 cgtacatggc gccattcgca gtcgagagaa atgtaaacaa atgtaaagat gcagtattaa
      541 atcataactg cataaaatta actgtagtac atactgtact actgtaataa tttcgtagcc
      601 acctcctgtt gctattgcgg tgagctcaag tgttgcgagt atccgcttaa aacgccgtgt
      661 gacgctaatc atctccgcgt gagcagttcg tctctccagt aaattgcgta tcgcagtaaa
      721 aagtgatctc tcgcggttct cgcgtatttt tcatcgtgtt tagtgcaata ccgtaaacct
      781 tgaataacac caccggtgct agcaattcta ccgggtaggg gaggcgcttt tcccaaggca
      841 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
      901 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
      961 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
     1021 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
     1081 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
     1141 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
     1201 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
     1261 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag
     1321 ccaacgccac catggggacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     1381 tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     1441 ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     1501 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     1561 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     1621 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     1681 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     1741 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     1801 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     1861 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     1921 agcccggtgc catcgatgcc actaacttct ccctgttgaa acaagcaggg gatgtcgaag
     1981 agaatcccgg gccaatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2041 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2101 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2161 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2221 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2281 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2341 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2401 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2461 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     2521 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2581 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     2641 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2701 agctgtacaa gtaattaatt aagctcgctg atcagcctcg actgtgcctt ctagttgcca
     2761 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2821 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2881 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2941 tgctggggat gcggtgggct ctatggcttc tgaggcgcgc cctcgagtaa ataatcatca
     3001 tgccgtacag ttaataaact tatctgttgt gtatgtgtgt gtcttcgtgt gaaaatctaa
     3061 taactgtacg gcaagaactg tatgagacat ttttgtgtca tcatcatcat cgcctaagta
     3121 ttcatcgtgt agaacatcgt gtgcaagact tgtatggaag tggatagcct atccttacat
     3181 aggcataagg tgagtgatat ttaatataaa attaataatg tgttagtttt cttactgttt
     3241 tataactttg ctttcaaaga attacattac cgtacagtat gcctctctct ctcgtaattg
     3301 gagaaactgc gtatcagcct atcatcacag gtaagtggtt ttttaaaaaa tgtaacaatg
     3361 tttccaatac tgtattatga atatgactgt aatactgtat gccataaaaa ttttataacg
     3421 attcattcat tagtgtatag gctaggctac cgtgaagcaa tcgtatcgat tacactaggc
     3481 taccataaag caatcatatt gctgcttctt cattatcaat gcatgaatcg ttatacctgt
     3541 aaataaatat gaatttcttt ttcacattat cttttcattt ttgatgtcta gtgttagtaa
     3601 tacgtataac atctacagtg ttttgtatca tataagacaa tattgatgta ggtactgaca
     3661 gatgattcat cttgtaaaca gatgacgtaa acttacggta tcgataaata cagtacagta
     3721 ctgtaaatgt attttctctt ccttatgatt ttcttaataa cattttcttt tctctagctt
     3781 actttattgt aagaatacag tatataatac atataacata caaaatatgt gttaatcgac
     3841 tgtttatgtt atcggtaagg cttctggtca acagtaggct attagtagtt aagttttggg
     3901 ggagtcaaaa gttatacgcg gattttcgac tgcgtggggg gtcggcgccc ctaacccccg
     3961 cgttgttcaa gggtcaactg taagagacgg agtcactgcc aaccgagacg gtcatagctg
     4021 tttcctgtgt gccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4081 agcggtatca gctcactcaa aggcggtaat acggttaccc acagaatcag gggataacgc
     4141 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4201 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4261 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4321 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4381 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4441 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4501 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4561 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4621 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4681 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4741 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4801 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4861 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4921 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4981 aatcagtgag gcacctatct cagcgatctg tctctttcgt tcatccatag ttgcctgact
     5041 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5101 aataccgcgg gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5161 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5221 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5281 cgctacaggc atcgtggtat cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5341 ccaacgatca aggcgagtta catgatcccc catgttgcgc aaaaaagcgg ttagctcctt
     5401 cggtcctccg atcgttgtca gaagtaagtt ggccgccgtg ttatcactca tggttatggc
     5461 agcactacat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5521 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5581 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5641 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5701 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5761 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5821 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5881 gagcggatac atatttgaat gtatttag
//
