LOCUS       sequence-443385-        4738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-3_FR  Transposon without spacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4738)
  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 4738)
  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 w/o spacer
FEATURES             Location/Qualifiers
     source          1..4738
                     /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        1036..1535
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1071..1094)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1468..1487
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1549..1558
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1555..2154
                     /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     2054..2074
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2161..2217
                     /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(2218..2220,2221..2223,2224..2937)
                     /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(2218..2220,2221..2223,2224..2934)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2263..2284)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2524..2543)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2871..2892
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2959..2976)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2965..3189
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3442..3458)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3442..3458)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3470..3487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      3539..3570
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3571..3673
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3674..3742,3743..4534)
                     /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(3892..3911)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4535..4726
                     /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 gctagcaatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt
     1081 tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca
     1141 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta
     1201 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac
     1261 aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga
     1321 agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc
     1381 agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg
     1441 gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg
     1501 cgctgttctc ctcttcctca tctccgggcc tttcgacctg cagccaacgc caccatgggg
     1561 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc
     1621 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc
     1681 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     1741 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     1801 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     1861 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     1921 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     1981 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     2041 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     2101 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgccatcgat
     2161 gccactaact tctccctgtt gaaacaagca ggggatgtcg aagagaatcc cgggccaatg
     2221 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2281 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2341 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2401 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     2461 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2521 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2581 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2641 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2701 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2761 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2821 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2881 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatta
     2941 attaagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3001 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3061 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3121 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3181 gctctatggc ttctgaggcg cgccgggatc aaatacttat taccctcaat gaaaaacaaa
     3241 taaattaaaa taaattcttt aaagttattt tctggatttt ctttttgata ttctgtctct
     3301 ccatgttaga atacatctac cattaaaagt atagaatgat catgtcttta ttagtggaca
     3361 aaccaacaaa atcagcaagg gatcaaatac tttttactct cactgtaggg cccagagacg
     3421 gagtcactgc caaccgagac ggtcatagct gtttcctgtg tgccgcttcc tcgctcactg
     3481 actcgctgcg ctcggtcgtt actgaccatt taaatcatac ctgacctcca tagcagaaag
     3541 tcaaaagcct ccgaccggag gcttttgact tgatcggcac gtaagaggtt ccaactttca
     3601 ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc
     3661 taaggaagct aaaatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3721 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3781 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3841 gagtttacgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3901 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3961 tcagaatgac ttggttgagt actcaccagt cacagaaaag catctcacgg atggcatgac
     4021 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4081 tctggcaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4141 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4201 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4261 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4321 atcacttctg cgctcggccc tcccggctgg ctggtttatt gctgataaat ctggagccgg
     4381 tgagcgtggg tcccgcggta tcattgcaga actggggcca gatggtaagc cctcccgcat
     4441 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4501 tgagataggt gcctcactga ttaagcattg gtaagtgacc aaacaggaaa aaaccgccct
     4561 taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct
     4621 ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta
     4681 ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgatgag ggcccaaa
//
