LOCUS       sequence-443594-        4611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-2_ST Transposon without spacer and inner DRs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4611)
  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 4611)
  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_ST DRiKO
FEATURES             Location/Qualifiers
     source          1..4611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      319..328
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(342..841)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(390..409)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     783..806
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(1060..1077)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1060..1076)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1086..1104)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1172..1276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1277..1345,1346..2137)
                     /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     complement(1495..1514)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2308..2896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2797..2816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3050..3067
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3079..3095
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3079..3095
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3299..3523
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3512..3529
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(3551..4264,4265..4267,4268..4270))
                     /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             complement(join(3554..4264,4265..4267,4268..4270))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3596..3617)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3945..3964
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4204..4225
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(4271..4327)
                     /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             complement(join(4334..4611,1..322))
                     /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     complement(4414..4434)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca
       61 ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg
      121 cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg
      181 tgacccgctc gatgtggcgg tccgggtcga cggtgtggcg cgtggcgggg tagtcggcga
      241 acgcggcggc gagggtgcgt acggcccggg ggacgtcgtc gcgggtggcg aggcgcaccg
      301 tgggcttgta ctcggtcccc atggtggcgt tggctgcagg tcgaaaggcc cggagatgag
      361 gaagaggaga acagcgcggc agacgtgcgc ttttgaagcg tgcagaatgc cgggcctccg
      421 gaggaccttc gggcgcccgc cccgcccctg agcccgcccc tgagcccgcc cccggaccca
      481 ccccttccca gcctctgagc ccagaaagcg aaggagcaaa gctgctattg gccgctgccc
      541 caaaggccta cccgcttcca ttgctcagcg gtgctgtcca tctgcacgag actagtgaga
      601 cgtgctactt ccatttgtca cgtcctgcac gacgcgagct gcggggcggg ggggaacttc
      661 ctgactaggg gaggagtaga aggtggcgcg aaggggccac caaagaacgg agccggttgg
      721 cgcctaccgg tggatgtgga atgtgtgcga ggccagaggc cacttgtgta gcgccaagtg
      781 cccagcgggg ctgctaaagc gcatgctcca gactgccttg ggaaaagcgc ctcccctacc
      841 cggtagaatt gctagcgcca aaatcatcac aattaaaaga accaaagact taaactactt
      901 cagtctgtgt gcattgaatt tatttaatac acgagtttca caatttgagt tgaattactg
      961 aaataaatga acttttccac gacattctaa tttattgaga tgcacctgta caattgagag
     1021 acgcaagaca ctgcggatcg agacgcattg cgtggttcga ctggccgtcg ttttacaacg
     1081 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1141 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1201 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1261 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1321 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1381 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1441 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1501 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1561 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1621 gacagtaaga gaattatgta gtgctgccat aaccatgagt gataacacgg cggccaactt
     1681 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcgca acatggggga
     1741 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1801 gcgtgatacc acgatgcctg tagcgatggc aacaacgttg cgcaaactat taactggcga
     1861 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1921 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1981 cggtgagcgt gggtcccgcg gtattattgc agcactgggg ccagatggta agccctcccg
     2041 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa agagacagat
     2101 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2161 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2221 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2281 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2341 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2401 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2461 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2521 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2581 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2641 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2701 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2761 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2821 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2881 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2941 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtgggta accgtattac
     3001 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3061 gagcgaggaa gcggcacaca ggaaacagct atgaccgtct cggttggcag tgactccgtc
     3121 tctgggccct acaggtgcat ctcaataaat tagaatgtcg tggaaaagtt catttatttc
     3181 agtaattcaa ctcaaattgt gaaactcgtg tattaaataa attcaatgca cacagactga
     3241 agtagtttaa gtctttggtt cttttaattg tgatgatttt ggcggcgcgc ctcagaagcc
     3301 atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg
     3361 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
     3421 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
     3481 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcga
     3541 gcttaattaa ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     3601 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggg
     3661 tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     3721 ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca
     3781 ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt
     3841 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     3901 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     3961 cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga
     4021 agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg
     4081 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg
     4141 tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt
     4201 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     4261 tgctcaccat tggcccggga ttctcttcga catcccctgc ttgtttcaac agggagaagt
     4321 tagtggcatc gatggcaccg ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca
     4381 cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg ttgcggggcg
     4441 cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact ccggggagca
     4501 cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gacgccgacg gtggccagga
     4561 accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt t
//
