LOCUS       sequence-442198-        4919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-8B_DR Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4919)
  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 4919)
  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-Tc1-8B_DR
FEATURES             Location/Qualifiers
     source          1..4919
                     /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        1205..1704
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1240..1263)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1637..1656
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1718..1727
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1724..2323
                     /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     2223..2243
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2330..2386
                     /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(2387..2389,2390..2392,2393..3106)
                     /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(2387..2389,2390..2392,2393..3103)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2432..2453)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2693..2712)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3040..3061
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3128..3145)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3134..3358
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3623..3639)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3623..3639)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3651..3668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      3720..3751
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3752..3854
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3855..3923,3924..4715)
                     /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(4073..4092)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4716..4907
                     /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 tcatatgtac agcggggaaa ataagtattt
      841 gacacatcag catttttatc agtaagggga tttctaagtg ggctactgac acaaaattcc
      901 taccagatgt agccatcaag ccaaatattg aattcataca aagaaatcag aacatttaag
      961 tatacaagtt gagtcataat aaataaagtg aaatgacaca gggaataagt attgaacaca
     1021 tgaagataac aaggtgcaaa atggcataga aagtcaggag atctgtcagt attgagagaa
     1081 aaaccctgct ccctatcagt actaattgat atcagctgct ttagtcctaa ttgatggcct
     1141 ataaaggctt ctcattactc aggaggcaca caggaaagac ttcaccggtg ctagcaattc
     1201 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     1261 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     1321 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     1381 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     1441 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     1501 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     1561 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg
     1621 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     1681 tcttcctcat ctccgggcct ttcgacctgc agccaacgcc accatgggga ccgagtacaa
     1741 gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc
     1801 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg
     1861 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     1921 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     1981 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     2041 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     2101 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     2161 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     2221 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     2281 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gccatcgatg ccactaactt
     2341 ctccctgttg aaacaagcag gggatgtcga agagaatccc gggccaatgg tgagcaaggg
     2401 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2461 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2521 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2581 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2641 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2701 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2761 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2821 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     2881 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     2941 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     3001 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3061 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaattaa ttaagctcgc
     3121 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3181 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3241 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3301 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3361 tctgaggcgc gccctcgagg taaagtgtgt tcaatactta ttccctgtgt catttcactt
     3421 tatttattat gactcaactt gtatacttaa atgttctgat ttctttgtat gaattcaata
     3481 tttggcttga tggctacatc tggtaggaat tttgtgtcag tagcccactt agaaatcccc
     3541 ttactgataa aaatgctgat gtgtcaaata cttattttcc ccgctgtagg gcccagagac
     3601 ggagtcactg ccaaccgaga cggtcatagc tgtttcctgt gtgccgcttc ctcgctcact
     3661 gactcgctgc gctcggtcgt tactgaccat ttaaatcata cctgacctcc atagcagaaa
     3721 gtcaaaagcc tccgaccgga ggcttttgac ttgatcggca cgtaagaggt tccaactttc
     3781 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
     3841 ctaaggaagc taaaatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3901 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3961 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4021 agagtttacg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4081 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4141 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatctcacg gatggcatga
     4201 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4261 ttctggcaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4321 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4381 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4441 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4501 gatcacttct gcgctcggcc ctcccggctg gctggtttat tgctgataaa tctggagccg
     4561 gtgagcgtgg gtcccgcggt atcattgcag aactggggcc agatggtaag ccctcccgca
     4621 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4681 ctgagatagg tgcctcactg attaagcatt ggtaagtgac caaacaggaa aaaaccgccc
     4741 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc
     4801 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt
     4861 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgatga gggcccaaa
//
