LOCUS       sequence-443616-        7869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-2_ST Transposon with 5 kb cargo gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7869)
  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 7869)
  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-5 kb
FEATURES             Location/Qualifiers
     source          1..7869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    239..463
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     452..469
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(491..1204,1205..1207,1208..1210))
                     /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(494..1204,1205..1207,1208..1210))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(536..557)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     885..904
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1144..1165
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1211..1267)
                     /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(1274..1873)
                     /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(1354..1374)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     regulatory      1870..1879
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1893..2392)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1941..1960)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     2334..2357
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(2728..2745)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2728..2744)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2754..2772)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2840..2944
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2945..3013,3014..3805)
                     /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(3163..3182)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3976..4564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4465..4484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4718..4735
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4747..4763
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4747..4763
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5213..5294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5250..5269)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6837..6860)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      6860..6869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(join(6884..7869,1..193))
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6893..7831
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(6916..6936)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 tcacggcgac tactgcactt atatacggtt ctcccccacc ctcgggaaaa aggcggagcc
       61 agtacacgac atcactttcc cagtttaccc cgcgccacct tctctaggca ccgggtcaat
      121 tgccgacccc tccccccaac ttctcgggga ctgtgggcga tgtgcgctct gcccactgac
      181 gggcaccgga gcctcacgtt aattaaccaa ggtacccatt ggcggccgcc ctcagaagcc
      241 atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg
      301 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
      361 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
      421 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcga
      481 gcttaattaa ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
      541 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggg
      601 tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
      661 ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca
      721 ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt
      781 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
      841 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
      901 cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga
      961 agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg
     1021 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg
     1081 tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt
     1141 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     1201 tgctcaccat tggcccggga ttctcttcga catcccctgc ttgtttcaac agggagaagt
     1261 tagtggcatc gatggcaccg ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca
     1321 cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg ttgcggggcg
     1381 cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact ccggggagca
     1441 cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gacgccgacg gtggccagga
     1501 accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg
     1561 ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac accgcccccg
     1621 cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca
     1681 ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct
     1741 cgatgtggcg gtccgggtcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg
     1801 cgagggtgcg tacggcccgg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt
     1861 actcggtccc catggtggcg ttggctgcag gtcgaaaggc ccggagatga ggaagaggag
     1921 aacagcgcgg cagacgtgcg cttttgaagc gtgcagaatg ccgggcctcc ggaggacctt
     1981 cgggcgcccg ccccgcccct gagcccgccc ctgagcccgc ccccggaccc accccttccc
     2041 agcctctgag cccagaaagc gaaggagcaa agctgctatt ggccgctgcc ccaaaggcct
     2101 acccgcttcc attgctcagc ggtgctgtcc atctgcacga gactagtgag acgtgctact
     2161 tccatttgtc acgtcctgca cgacgcgagc tgcggggcgg gggggaactt cctgactagg
     2221 ggaggagtag aaggtggcgc gaaggggcca ccaaagaacg gagccggttg gcgcctaccg
     2281 gtggatgtgg aatgtgtgcg aggccagagg ccacttgtgt agcgccaagt gcccagcggg
     2341 gctgctaaag cgcatgctcc agactgcctt gggaaaagcg cctcccctac ccggtagaat
     2401 tgctagcacc ggttttgaag gctcaggaaa cctttgcagg tgttttgagt tgattagctg
     2461 attggcatgt caccatattc taatttgttg agatagtgaa ttggtgggtt tttgttaaat
     2521 gtgagccaaa atcatcacaa ttaaaagaac caaagactta aactacttca gtctgtgtgc
     2581 attgaattta tttaatacac gagtttcaca atttgagttg aattactgaa ataaatgaac
     2641 ttttccacga cattctaatt tattgagatg cacctgtaca attgagagac gcaagacact
     2701 gcggatcgag acgcattgcg tggttcgact ggccgtcgtt ttacaacgaa agggcctcgt
     2761 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     2821 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2881 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2941 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3001 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3061 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3121 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3181 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3241 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3301 attatgtagt gctgccataa ccatgagtga taacacggcg gccaacttac ttctgacaac
     3361 gatcggagga ccgaaggagc taaccgcttt tttgcgcaac atgggggatc atgtaactcg
     3421 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgataccac
     3481 gatgcctgta gcgatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3541 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3601 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3661 gtcccgcggt attattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3721 ctacacgacg gggagtcagg caactatgga tgaacgaaag agacagatcg ctgagatagg
     3781 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3841 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3901 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3961 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4021 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4081 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4141 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4201 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4261 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4321 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4381 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4441 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4501 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4561 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4621 catgttcttt cctgcgttat cccctgattc tgtgggtaac cgtattaccg cctttgagtg
     4681 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4741 ggcacacagg aaacagctat gaccgtctcg gttggcagtg actccgtctc tgggccctac
     4801 aggtgcatct caataaatta gaatgtcgtg gaaaagttca tttatttcag taattcaact
     4861 caaattgtga aactcgtgta ttaaataaat tcaatgcaca cagactgaag tagtttaagt
     4921 ctttggttct tttaattgtg atgattttgg ctcacattta acaaaaaccc accaattcac
     4981 tatctcaaca aattagaata cttcataaga ccaataaaaa aaacattttt agtgaattgt
     5041 tggccttctg gaaagtatgt tcatttactg tacatgtacc tcgagggcca ggaagcttcc
     5101 ctttggtctc cccctcttaa gtaccacatt tgtagaggtt ttacttgctt taaaaaacct
     5161 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     5221 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5281 atttttttca ctgcattcta gttgtggttt gtccaaactc aatcgatgcg agggggcagg
     5341 gcctgcatgt gaagggcgtc gtaggtgtcc ttggtggctg tactgagacc ctggtaaagg
     5401 ccatcgtgcc ccttgcccct ccggcgctcg cctttcatcc caatctcact gtaggcctcc
     5461 gccatcttat ctttctgcag ttcattgtac aggccttcct gagggttctt ccttctcggc
     5521 tttcccccca tctcagggtc ccggccacgt ctcttgtcca aaacatcgta ctcctctctt
     5581 cgtcctagat tgagctcgtt atagagctgg ttctggccct gctggtacgc gggggcgtct
     5641 gcgctcctgc tgaacttcac tctggagcga taggctgcga agtcgcgtgg tggggcatag
     5701 ggctggtaat gcttgcgggt gggcccgggg cggcggggag tcatgttcat gtagtcactg
     5761 tgcaggagcc tgctcctctt actcctcacc cagaaaataa taaaggccac tgttactagc
     5821 aagctatagc aagccaggac tcccccaacc accaccagca cccaaaaatc acaggcgaag
     5881 tccagccccc tcgtgtgcac tgcgcccccc gccgctggcc ggcacgcctc tgggcgcagg
     5941 gacaggggct gcgacgcgat ggtgggcgcc ggtgttggtg gtcgcggcgc tggcgtcgtg
     6001 gtcagcactg tcagttgcgt gcctccaccg aaaacagcgg ctgaactatg ccaaatcata
     6061 cagtaataat cggcctcgtc ctcggacctg agacctgaga tgagcagcac acccgcattg
     6121 gcgcttgcgt ctttagatcc actaaacctt gatggaactc cagatccctg ctgcttatct
     6181 gagtcgctct tgtaattgag aaggtattgg ggaggagaac ctggtttttg ctgataccag
     6241 tagatcctgt aaggccctac attgattcct gacctcagtg tacacgtcaa agaagcggag
     6301 gcaccggggc ttgcactgag ggaggatgat tgcgtgagaa caggttggct acccccccct
     6361 ccggacccgc ccccaccgga acctccaccg cctgagccca ggatcccact ggacacggtg
     6421 acggttgtac cttgtcccca aacgtccata ccatagtagt aagtcatcat accccgcgca
     6481 caatagtaaa cagctgtgtc ttcgggagtt acactattaa gctgaagaga gaactggttt
     6541 ttggaagtat ctggatttat agacatcctg cttttcacac ttacagcata atcattatac
     6601 catttactgc ggtagtatgt tctaccgagc cattccaacc ctcgactggg ggactgcctg
     6661 atccaattcc atgtcgcgga gttactactc actgaatcac ctgatatggc acaggtaaga
     6721 ctaagagtct gggaaggagt gaccagaccg gggccagatt gttggagttg cacttgcggc
     6781 ctggcggcgt ggagcagcaa ggccagcggc aggagcaagg cggtcactgg taaggcctta
     6841 tcgtcatcgt ctttgtaatc catggtggcg accggtggat ccctcacgac acctgaaatg
     6901 gaagaaaaaa actttgaacc actgtctgag gcttgagaat gaaccaagat ccaaactcaa
     6961 aaagggcaaa ttccaaggag aattacatca agtgccaagc tggcctaact tcagtctcca
     7021 cccactcagt gtggggaaac tccatcgcat aaaacccctc cccccaacct aaagacgacg
     7081 tactccaaaa gctcgagaac taatcgaggt gcctggacgg cgcccggtac tcagtggagt
     7141 cacatgaagc gacggctgag gacggaaagg cccttttcct ttgtgtgggt gactcacccg
     7201 cccgctctcc cgagcgccgc gtcctccatt ttgagctccc tgcagcaggg ccgggaagcg
     7261 gccatctttc cgctcacgca actggtgccg accgggccag ccttgccgcc cagggcgggg
     7321 cgatacacgg cggcgcgagg ccaggcacca gagcaggccg gcgagcttga gactaccccc
     7381 gtccgattct cggtggccgc gctcgcaggc cccgcctcgc cgaacatgtg cgctgggacg
     7441 cacgggcccc gtcgccgccc gcggccccaa aaaccgaaat accagtgtgc agatcttggc
     7501 ccgcatttac aagactatct tgccagaaaa aaagcgtcgc agcaggtcat caaaaatttt
     7561 aaatggctag agacttatcg aaagcagcga gacaggcgcg aaggtgccac cagattcgca
     7621 cgcggcggcc ccagcgccca ggccaggcct caactcaagc acgaggcgaa ggggctcctt
     7681 aagcgcaagg cctcgaactc tcccacccac ttccaacccg aagctcggga tcaagaatca
     7741 cgtactgcag ccaggtggaa gtaattcaag gcacgcaagg gccataaccc gtaaagaggc
     7801 caggcccgcg ggaaccacac acggcactta cctgtgttct ggcggcaaac ccgttgcgaa
     7861 aaagaacgt
//
