LOCUS       sequence-443558-        7870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-2_PM Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7870)
  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 7870)
  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-Tc1-2_PM
FEATURES             Location/Qualifiers
     source          1..7870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..112
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     121..138
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     122..138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2520..3019
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2555..2578)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2952..2971
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      3033..3042
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3039..3638
                     /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     3538..3558
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3645..3701
                     /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(3702..3704,3705..3707,3708..4421)
                     /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(3702..3704,3705..3707,3708..4418)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3747..3768)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4008..4027)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4355..4376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4443..4460)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4449..4673
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5973..5989)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5973..5989)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6001..6018)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6172..6760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6252..6271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6931..7722,7723..7791))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLRNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERKRQIAEIGAS
                     LIKHW"
     primer_bind     7554..7573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7792..7870,1..26))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
       61 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
      121 tgtaaaacga cggccagtcg aaccacgcaa tgcgtctcga tccgcagtgt cttgcgtctc
      181 tcaattgtac agtgccgaga aaaagtttgt gaaccccaat gataattatg gaaattccat
      241 agttttacca tgatagcctt cttgaatcaa aaccggtctt ttcttaaata tccaataggg
      301 tttatattaa ccaattccat gtcttttgaa acaaaatgat gtatgaatta aacaaacaat
      361 gagtaattaa ggttcagggt atgtgaaaaa gtaagtgaac ccctggtttc attagctcaa
      421 ttaagggaat aattagaatc aggtgtttaa ataattaggt agatcttcag gggtgagttt
      481 gggaggcccc accctatata aagatcagaa actttgtgag tttggtcttc accatacagg
      541 tgtgtggaaa caagtcatgc cacgatcaaa ataaatctct gaggacctca gaaaaagagt
      601 tattgatgct catcagtccg gaaagggtta caaaaccatt tctaaggata ccggtcaatt
      661 gtacaggcat acctcgtttt attgcgcttc gctttattgc gcttcgcaga tattgcgttt
      721 tttacaaatt gaaggtttgt ggcaaccctg cgtcgagcaa gtctatcggc gccatttttc
      781 caacagcatg tgctcacttc gtgtctctgt gtcacatttt ggtaattctc gcaatatttc
      841 aaactttttc attattatta tatctgttat ggtgatctgt gatcagtgat ctttgatgtt
      901 actattgtaa ttgttttggg gcgccacgaa ccgcgcccat ataagacggc gaacttaatc
      961 gataaatgtt gtgtgtgttc tgactgctcc accgaccggc cgttccccca tctctctccc
     1021 tctcctcggg cctccctatt ccctgagaca caacaatatt gaaattaggc caattaataa
     1081 ccctacaacc ggtcaattgt ttccttgcag ggctgcccag aggggggggg gggcaactgg
     1141 ggcaatttgc cccgggcccc tgcaaagcaa gggccccctg cctcagctcc tgtgtgtacc
     1201 gttggcccct gccctgcgac atcgctcacc agaattcttg gtattttgtt ttcatgaaaa
     1261 agaggaaatt ggtgtattag agtgtaagat gacactgaat cattctcaac atgtaattgt
     1321 gacattatgg gacccgtccc cgaccatgaa gtaaatatta tacatagttg tgtgtgaaac
     1381 agatgctaat ttgaggctta agtgattgtt gcattgatgt ccaaggggga gtctaaacca
     1441 atcacggtaa ggtaacatct ttctttatct ctgggcagca atagagacaa gattgatcag
     1501 gcaggcttat gattggccta gcctagggcc taggccaagt agccctggtt acagagtctg
     1561 ttgtgaggag agaaggaaca gggaaggctt taaggaggga tctgtggcag agccggcctg
     1621 ttgtcacaga aagacctcca ggggaaagag ttcaagtcgt gagtgaatgt cagctctgtg
     1681 ggttcttaag ggccaaagtc agaaatgaat ttgcaagaat ataataatta aaatgatata
     1741 tattgttttc tttatatttg ccaaggaaaa taacaacatt agacataaca catattcagg
     1801 ttcacatatt tccccacttc aaaaagtact taaatgcaaa gtgtctatat aactcttggg
     1861 gaaaaatatt aaagtccttc ttgattgaag aagcctgtgc tccaaactgc aaatcttgct
     1921 ttcagtattg cagagatgac ttgctctctt ctggtccttc aaacagcagc agttgcttac
     1981 taaggcccat aagaatatga acagtctcac aatgccctta atcaaggggt taaaggctaa
     2041 atatacaagt ttattctcca gcaacagtac aaagacaatc ctccttcctc tgtctactct
     2101 ctctgcagtt actttctttt tccttaaaca ggggcagggc ctgcttatcc tctgccacac
     2161 ccacaagcct gaaccaagta caacccagct tatgggggtt acatgagcac cagctaagat
     2221 ttccaggcaa ggtcccactg gagagtaatt ttctgtggca gttaccacag ggaaaaggcc
     2281 taggagtggt gtgaggccag gccttaaggg aggaatcctt ccttcctgag tagtggagga
     2341 ggccagccag caggccacaa ccagaagacc tgtgagactg agccgggccc caggaaactg
     2401 gtaggagaga gaagctctgc cagaaggggt ttggaatttg gaggtgctca ggagaagtac
     2461 aaccctattc ctatcccctt atccccgtgt aagaaaaaac cggtgctagc aattctaccg
     2521 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg
     2581 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg
     2641 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag
     2701 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg
     2761 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg
     2821 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg
     2881 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
     2941 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     3001 ctcatctccg ggcctttcga cctgcagcca acgccaccat ggggaccgag tacaagccca
     3061 cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt
     3121 tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca
     3181 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     3241 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     3301 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     3361 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     3421 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     3481 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     3541 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     3601 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccat cgatgccact aacttctccc
     3661 tgttgaaaca agcaggggat gtcgaagaga atcccgggcc aatggtgagc aagggcgagg
     3721 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3781 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     3841 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     3901 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3961 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     4021 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     4081 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     4141 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     4201 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     4261 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     4321 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     4381 ccgccgggat cactctcggc atggacgagc tgtacaagta attaattaag ctcgctgatc
     4441 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4501 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4561 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4621 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     4681 ggcgcgccct cgagactttc ctcttctcat tttgttctta agttcctaac tttgtagcat
     4741 aagtttaata aatgtttcac ttgcaaactg ttaatgtcat tttgctttag taacctgtct
     4801 ccaaggtaaa ttctctctgc ctagccctcc ttgccacggg aacaagcggg gaaagcccac
     4861 ttgttcccac ttcccagggc aagggaagga ggagccgcat gtgagaattc tcacatgcga
     4921 tgcgtgcgtg caagaattct cgcacgtggt cccccagctt tgtacacaca cacacgggtc
     4981 cttgtagggg gcagtggtgg ggcccccaac atcacacctg ccccaggccc cataaaccct
     5041 ctgggcagcc ctgtttcctt ggggcccctc gagacggatg aggagttgct tcttatggat
     5101 gagcaaagaa agtggtttct tgagatggaa tctactcctg gtgaagatgc tgtgaacatt
     5161 gttgaaatga caacaaagga tttagaatat tacataaact tagttgataa agcagcggca
     5221 gggtttgaga ggattgactc caattttgaa agaagttcta ctgtgggtaa aatgctatca
     5281 aacagcatcg catgctacag agaaatcttt cgtgaaagga agagtcaatc gatgcggcaa
     5341 acttcattgt tgtcttattt taagaaattg ccacagccac cccaaccttc agcaaccacc
     5401 accctgatca gtcagcagcc atcaacatcg aggcaagacc ctccaccagc aaaaagatta
     5461 cgactcgctg aaggctcaga tgatcgttag cattttttag caataaagta tttttaaatt
     5521 aaggtatgta cattgttttt tagacataat gctattgcac acttaataga ctacagtata
     5581 gtgtaaacat aacttttata tgcactggga aaccaaaaaa ttcgtgtgac tcgctttatt
     5641 gcgatattcg ctttattgcg gtggtctgga accgaacccg caatatctcc gaggtatgcc
     5701 tgtagggccc ctcgagatct aaaggttcac atactttttc acacgtggtt attgaatgta
     5761 gaatcatttg aggataaata aatgttgaaa gagtatcatg tttttgtgtc atttgtttaa
     5821 tcaggttatc tttatctatt attaggactt agattaagat ctaataacat tttaggtttg
     5881 aaatatgtaa aatatgtgaa aatcctaagg ggttcacaaa ctttttctcg gcactgtagg
     5941 gcccagagac ggagtcactg ccaaccgaga cggtcatagc tgtttcctgt gtgccgcttc
     6001 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6061 aaaggcggta atacggttac ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6121 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6181 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6241 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6301 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6361 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6421 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6481 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6541 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6601 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6661 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6901 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6961 ctcagcgatc tgtctctttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7021 tacgatacgg gagggcttac catctggccc cagtgctgca ataataccgc gggacccacg
     7081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc atcgctacag gcatcgtggt
     7261 atcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7321 tacatgatcc cccatgttgc gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7381 cagaagtaag ttggccgccg tgttatcact catggttatg gcagcactac ataattctct
     7441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7861 atgtatttag
//
