LOCUS       sequence-442190-        4762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-1_AG Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4762)
  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 4762)
  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-Tc1-1_AG
FEATURES             Location/Qualifiers
     source          1..4762
                     /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        1112..1611
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1147..1170)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1544..1563
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1625..1634
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1631..2230
                     /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     2130..2150
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2237..2293
                     /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(2294..2296,2297..2299,2300..3013)
                     /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(2294..2296,2297..2299,2300..3010)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2339..2360)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2600..2619)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2947..2968
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3035..3052)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3041..3265
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3466..3482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3466..3482)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3494..3511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      3563..3594
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3595..3697
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3698..3766,3767..4558)
                     /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(3916..3935)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4559..4750
                     /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 taagctttac actggtggac ataaaaatag
      841 gaaaaaaaaa ttttgtgtgt tttttttgca tacactaggt gtgtcatcgc caacagttcg
      901 aggcgtactg aatttgcaat agccgaattt tgccttttat actctcattt ttggtgcgct
      961 acttatctga gttttgagtg ccggcagcgt tttgcggcag tataaaagga gagccaaacc
     1021 gtgttgtgct tcattcttcc atcagtggtc aaggtgaaag gttgctattt aaaaattcca
     1081 caaaatcatc accggtgcta gcaattctac cgggtagggg aggcgctttt cccaaggcag
     1141 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
     1201 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
     1261 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
     1321 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc
     1381 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
     1441 ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg
     1501 ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa
     1561 aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc
     1621 caacgccacc atggggaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     1681 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     1741 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     1801 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     1861 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     1921 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     1981 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     2041 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     2101 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     2161 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     2221 gcccggtgcc atcgatgcca ctaacttctc cctgttgaaa caagcagggg atgtcgaaga
     2281 gaatcccggg ccaatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2341 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2401 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2461 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2521 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2581 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2641 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2701 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2761 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2821 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2881 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2941 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3001 gctgtacaag taattaatta agctcgctga tcagcctcga ctgtgccttc tagttgccag
     3061 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3121 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3181 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3241 gctggggatg cggtgggctc tatggcttct gaggcgcgcc ctcgagaatg tgttttcgct
     3301 cagtggaaca ccgcaacacc tcctcccaac aaccacttaa acagtccttt tcagggctac
     3361 caaatatttc tgacaagaac tatgtctttc ggtcacagaa aaaagttcct atttttatgt
     3421 ccaccagtgt agggcccaga gacggagtca ctgccaaccg agacggtcat agctgtttcc
     3481 tgtgtgccgc ttcctcgctc actgactcgc tgcgctcggt cgttactgac catttaaatc
     3541 atacctgacc tccatagcag aaagtcaaaa gcctccgacc ggaggctttt gacttgatcg
     3601 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
     3661 tgagttatcg agattttcag gagctaagga agctaaaatg agtattcaac atttccgtgt
     3721 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3781 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3841 tctcaacagc ggtaagatcc ttgagagttt acgccccgaa gaacgttttc caatgatgag
     3901 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3961 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4021 aaagcatctc acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4081 tgataacact gcggccaact tacttctggc aacgatcgga ggaccgaagg agctaaccgc
     4141 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4201 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4261 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4321 gatggaggcg gataaagttg caggatcact tctgcgctcg gccctcccgg ctggctggtt
     4381 tattgctgat aaatctggag ccggtgagcg tgggtcccgc ggtatcattg cagaactggg
     4441 gccagatggt aagccctccc gcatcgtagt tatctacacg acggggagtc aggcaactat
     4501 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaagt
     4561 gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac
     4621 gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct
     4681 tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga
     4741 aaacctctga tgagggccca aa
//
