LOCUS       sequence-443596-        4898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-3_FR Transposon with copGFP mammalian 
            expression cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4898)
  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 4898)
  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: pQLL-Tc1-3_FR[EF1-alpha-copGFP]
FEATURES             Location/Qualifiers
     source          1..4898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      12..39
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(51..638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(131..150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(660..689)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     primer_bind     724..742
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     751..768
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     752..768
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1208..2386
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1439..2377
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2334..2354
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2428..3093
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             2428..3093
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     primer_bind     complement(3107..3126)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    3136..3217
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3161..3180
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3602..3618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3602..3618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3630..3647)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      3699..3730
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3731..3833
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3834..3902,3903..4694)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAELGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4052..4071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4695..4886
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tgtaatcacc tggctcacct tcgggtgggc ctttctgcgt tgctggcgtt tttccatagg
       61 ctccgccccc ctgacgagca tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg
      121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      481 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac ctcggaaaaa
      541 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg attttctacc
      661 gaagaaaggc ccacccgtga aggtgagcca gtgagttgat tgcagtccag ttacgctgga
      721 gtcaataggc gtatcacgag gccctttcgt tgtaaaacga cggccagtcg aaccacgcaa
      781 tgcgtctcga tccgcagtgt cttgcgtctc tggtacctac agtgagagta aaaagtattt
      841 gatcccttgc tgattttgtt ggtttgtcca ctaataaaga catgatcatt ctatactttt
      901 aatggtagat gtattctaac atggagagac agaatatcaa aaagaaaatc aagaaaataa
      961 ctttaaagaa tttattttaa tttatttgtt tttcattgag ggaaataagt atttgatccc
     1021 ctagtatgca ttaggagttc tggctttcac agaccagtta gacgctccca aacaacttgt
     1081 tacctgaatt gaagacacca ggtctaacta atcacctgta tgtaagacaa ctgttcacag
     1141 aatcagacaa tcagacagat tccaaactct ccaccaccgg tgctagaggc attgattatt
     1201 gactagtggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     1261 gttgggggga ggggtcggca attgacccgg tgcctagaga aggtggcgcg gggtaaactg
     1321 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     1381 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt
     1441 aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc
     1501 ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag
     1561 tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga
     1621 ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct
     1681 cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt
     1741 tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt
     1801 tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg
     1861 cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctcgcc ggcctgctct
     1921 ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc
     1981 ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa
     2041 atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc
     2101 ctttccgtcc tcagccgtcg cttcatgtga ctccactgag taccgggcgc cgtccaggca
     2161 cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta
     2221 tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt
     2281 gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc
     2341 tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaggga tccggtaccg
     2401 aagccgctag cgctaccggt cgccaccatg cccgccatga agatcgagtg ccgcatcacc
     2461 ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccgagcag
     2521 ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg
     2581 ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag
     2641 aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac
     2701 gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc
     2761 ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag
     2821 atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg
     2881 ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac
     2941 agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg
     3001 ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac
     3061 cagcacgcct tcaagacccc catcgccttc gccagatctc gagctcgatg agtttggaca
     3121 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3181 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3241 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     3301 atgtggtact taagaggggg agaccaaagg gcgagggcgc gccctcgagg tgcttttgga
     3361 tagagggatc aaatacttat taccctcaat gaaaaacaaa taaattaaaa taaattcttt
     3421 aaagttattt tctggatttt ctttttgata ttctgtctct ccatgttaga atacatctac
     3481 cattaaaagt atagaatgat catgtcttta ttagtggaca aaccaacaaa atcagcaagg
     3541 gatcaaatac tttttactct cactgtaggg cccagagacg gagtcactgc caaccgagac
     3601 ggtcatagct gtttcctgtg tgccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3661 actgaccatt taaatcatac ctgacctcca tagcagaaag tcaaaagcct ccgaccggag
     3721 gcttttgact tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
     3781 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatgagta
     3841 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3901 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3961 gttacatcga actggatctc aacagcggta agatccttga gagtttacgc cccgaagaac
     4021 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4081 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4141 actcaccagt cacagaaaag catctcacgg atggcatgac agtaagagaa ttatgcagtg
     4201 ctgccataac catgagtgat aacactgcgg ccaacttact tctggcaacg atcggaggac
     4261 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4321 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4381 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4441 aacaattaat agactggatg gaggcggata aagttgcagg atcacttctg cgctcggccc
     4501 tcccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tcccgcggta
     4561 tcattgcaga actggggcca gatggtaagc cctcccgcat cgtagttatc tacacgacgg
     4621 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4681 ttaagcattg gtaagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca
     4741 gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag
     4801 acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt
     4861 tcggtgatga cggtgaaaac ctctgatgag ggcccaaa
//
