LOCUS       sequence-445654-        7921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-3_FR Transposon with 5 kb cargo gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7921)
  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 7921)
  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-5 kb
FEATURES             Location/Qualifiers
     source          1..7921
                     /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        1197..1696
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1232..1255)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1629..1648
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1710..1719
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1716..2315
                     /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     2215..2235
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2322..2378
                     /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(2379..2381,2382..2384,2385..3098)
                     /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(2379..2381,2382..2384,2385..3095)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2424..2445)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2685..2704)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3032..3053
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3120..3137)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3126..3350
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3390..4568
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3621..4559
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4516..4536
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      4583..4592
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4592..4615
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    6158..6239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     6183..6202
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6625..6641)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6625..6641)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6653..6670)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      6722..6753
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        6754..6856
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6857..6925,6926..7717)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7075..7094)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             7718..7909
                     /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 tgctagcaat tctaccgggt
     1201 aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac
     1261 ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca
     1321 accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa
     1381 gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct
     1441 cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg
     1501 cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg
     1561 gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg
     1621 aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc
     1681 atctccgggc ctttcgacct gcagccaacg ccaccatggg gaccgagtac aagcccacgg
     1741 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg
     1801 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg
     1861 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     1921 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     1981 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     2041 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     2101 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     2161 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     2221 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     2281 cgcgcacctg gtgcatgacc cgcaagcccg gtgccatcga tgccactaac ttctccctgt
     2341 tgaaacaagc aggggatgtc gaagagaatc ccgggccaat ggtgagcaag ggcgaggagc
     2401 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2461 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     2521 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     2581 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2641 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2701 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2761 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2821 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2881 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2941 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     3001 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     3061 ccgggatcac tctcggcatg gacgagctgt acaagtaatt aattaagctc gctgatcagc
     3121 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3181 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3241 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3301 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggg
     3361 cggccgccat atgggtacca ttaacgtgag gctccggtgc ccgtcagtgg gcagagcgca
     3421 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaccc ggtgcctaga
     3481 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     3541 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     3601 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
     3661 cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg
     3721 atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc
     3781 cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct
     3841 ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt
     3901 gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc
     3961 tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc
     4021 gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt
     4081 ctcaagctcg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct
     4141 gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg
     4201 gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt
     4261 cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccactg
     4321 agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt
     4381 taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg
     4441 aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg
     4501 gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg
     4561 tgtcgtgagg gatccaccgg tcgccaccat ggattacaaa gacgatgacg ataaggcctt
     4621 accagtgacc gccttgctcc tgccgctggc cttgctgctc cacgccgcca ggccgcaagt
     4681 gcaactccaa caatctggcc ccggtctggt cactccttcc cagactctta gtcttacctg
     4741 tgccatatca ggtgattcag tgagtagtaa ctccgcgaca tggaattgga tcaggcagtc
     4801 ccccagtcga gggttggaat ggctcggtag aacatactac cgcagtaaat ggtataatga
     4861 ttatgctgta agtgtgaaaa gcaggatgtc tataaatcca gatacttcca aaaaccagtt
     4921 ctctcttcag cttaatagtg taactcccga agacacagct gtttactatt gtgcgcgggg
     4981 tatgatgact tactactatg gtatggacgt ttggggacaa ggtacaaccg tcaccgtgtc
     5041 cagtgggatc ctgggctcag gcggtggagg ttccggtggg ggcgggtccg gagggggggg
     5101 tagccaacct gttctcacgc aatcatcctc cctcagtgca agccccggtg cctccgcttc
     5161 tttgacgtgt acactgaggt caggaatcaa tgtagggcct tacaggatct actggtatca
     5221 gcaaaaacca ggttctcctc cccaatacct tctcaattac aagagcgact cagataagca
     5281 gcagggatct ggagttccat caaggtttag tggatctaaa gacgcaagcg ccaatgcggg
     5341 tgtgctgctc atctcaggtc tcaggtccga ggacgaggcc gattattact gtatgatttg
     5401 gcatagttca gccgctgttt tcggtggagg cacgcaactg acagtgctga ccacgacgcc
     5461 agcgccgcga ccaccaacac cggcgcccac catcgcgtcg cagcccctgt ccctgcgccc
     5521 agaggcgtgc cggccagcgg cggggggcgc agtgcacacg agggggctgg acttcgcctg
     5581 tgatttttgg gtgctggtgg tggttggggg agtcctggct tgctatagct tgctagtaac
     5641 agtggccttt attattttct gggtgaggag taagaggagc aggctcctgc acagtgacta
     5701 catgaacatg actccccgcc gccccgggcc cacccgcaag cattaccagc cctatgcccc
     5761 accacgcgac ttcgcagcct atcgctccag agtgaagttc agcaggagcg cagacgcccc
     5821 cgcgtaccag cagggccaga accagctcta taacgagctc aatctaggac gaagagagga
     5881 gtacgatgtt ttggacaaga gacgtggccg ggaccctgag atggggggaa agccgagaag
     5941 gaagaaccct caggaaggcc tgtacaatga actgcagaaa gataagatgg cggaggccta
     6001 cagtgagatt gggatgaaag gcgagcgccg gaggggcaag gggcacgatg gcctttacca
     6061 gggtctcagt acagccacca aggacaccta cgacgccctt cacatgcagg ccctgccccc
     6121 tcgcatcgat tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     6181 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     6241 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     6301 aaagcaagta aaacctctac aaatgtggta cttaagaggg ggagaccaaa gggaagcttc
     6361 ctggccctcg aggtgctttt ggatagaggg atcaaatact tattaccctc aatgaaaaac
     6421 aaataaatta aaataaattc tttaaagtta ttttctggat tttctttttg atattctgtc
     6481 tctccatgtt agaatacatc taccattaaa agtatagaat gatcatgtct ttattagtgg
     6541 acaaaccaac aaaatcagca agggatcaaa tactttttac tctcactgta gggcccagag
     6601 acggagtcac tgccaaccga gacggtcata gctgtttcct gtgtgccgct tcctcgctca
     6661 ctgactcgct gcgctcggtc gttactgacc atttaaatca tacctgacct ccatagcaga
     6721 aagtcaaaag cctccgaccg gaggcttttg acttgatcgg cacgtaagag gttccaactt
     6781 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     6841 agctaaggaa gctaaaatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6901 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6961 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     7021 tgagagttta cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     7081 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     7141 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctca cggatggcat
     7201 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7261 acttctggca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7321 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     7381 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     7441 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7501 aggatcactt ctgcgctcgg ccctcccggc tggctggttt attgctgata aatctggagc
     7561 cggtgagcgt gggtcccgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     7621 catcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     7681 cgctgagata ggtgcctcac tgattaagca ttggtaagtg accaaacagg aaaaaaccgc
     7741 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
     7801 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
     7861 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgat gagggcccaa
     7921 a
//
