LOCUS       sequence-443598-        7910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-1_PM Transposon with 5 kb cargo gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7910)
  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 7910)
  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-Tc1-1_PM-5 kb
FEATURES             Location/Qualifiers
     source          1..7910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    783..1007
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     996..1013
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(1035..1748,1749..1751,1752..1754))
                     /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(1038..1748,1749..1751,1752..1754))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1080..1101)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1429..1448
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1688..1709
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1755..1811)
                     /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(1818..2417)
                     /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(1898..1918)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     regulatory      2414..2423
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2437..2936)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2485..2504)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     2878..2901
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(3377..3394)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3377..3393)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3403..3421)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3489..3593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3594..3662,3663..4454)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3812..3831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4625..5213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5114..5133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5367..5384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5396..5412
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5396..5412
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5798..5879
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5835..5854)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(7422..7445)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      7445..7454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(join(7469..7910,1..737))
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(7478..7910,1..506)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(7501..7521)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 cacggcggcg cgaggccagg caccagagca ggccggcgag cttgagacta cccccgtccg
       61 attctcggtg gccgcgctcg caggccccgc ctcgccgaac atgtgcgctg ggacgcacgg
      121 gccccgtcgc cgcccgcggc cccaaaaacc gaaataccag tgtgcagatc ttggcccgca
      181 tttacaagac tatcttgcca gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg
      241 gctagagact tatcgaaagc agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg
      301 cggccccagc gcccaggcca ggcctcaact caagcacgag gcgaaggggc tccttaagcg
      361 caaggcctcg aactctccca cccacttcca acccgaagct cgggatcaag aatcacgtac
      421 tgcagccagg tggaagtaat tcaaggcacg caagggccat aacccgtaaa gaggccaggc
      481 ccgcgggaac cacacacggc acttacctgt gttctggcgg caaacccgtt gcgaaaaaga
      541 acgttcacgg cgactactgc acttatatac ggttctcccc caccctcggg aaaaaggcgg
      601 agccagtaca cgacatcact ttcccagttt accccgcgcc accttctcta ggcaccgggt
      661 caattgccga cccctccccc caacttctcg gggactgtgg gcgatgtgcg ctctgcccac
      721 tgacgggcac cggagcctca cgttaattaa ccaaggtacc cattggcggc cgccctcaga
      781 agccatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat cctccccctt
      841 gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc
      901 aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg
      961 cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg aggctgatca
     1021 gcgagcttaa ttaattactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     1081 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac
     1141 tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     1201 tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag
     1261 ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag
     1321 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     1381 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     1441 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     1501 aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc
     1561 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc
     1621 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg
     1681 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     1741 cccttgctca ccattggccc gggattctct tcgacatccc ctgcttgttt caacagggag
     1801 aagttagtgg catcgatggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg
     1861 ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg
     1921 ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg
     1981 agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc
     2041 aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc
     2101 gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc
     2161 cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc
     2221 cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc
     2281 cgctcgatgt ggcggtccgg gtcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg
     2341 gcggcgaggg tgcgtacggc ccgggggacg tcgtcgcggg tggcgaggcg caccgtgggc
     2401 ttgtactcgg tccccatggt ggcgttggct gcaggtcgaa aggcccggag atgaggaaga
     2461 ggagaacagc gcggcagacg tgcgcttttg aagcgtgcag aatgccgggc ctccggagga
     2521 ccttcgggcg cccgccccgc ccctgagccc gcccctgagc ccgcccccgg acccacccct
     2581 tcccagcctc tgagcccaga aagcgaagga gcaaagctgc tattggccgc tgccccaaag
     2641 gcctacccgc ttccattgct cagcggtgct gtccatctgc acgagactag tgagacgtgc
     2701 tacttccatt tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga acttcctgac
     2761 taggggagga gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg gttggcgcct
     2821 accggtggat gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc aagtgcccag
     2881 cggggctgct aaagcgcatg ctccagactg ccttgggaaa agcgcctccc ctacccggta
     2941 gaattgctag caccggtggt ggagagtttg gaatctgatt gattgattgc ttctgtggac
     3001 aggtgtcttt tatacaggta acaagctgag attaggagca ctccctttaa gagtgtgctc
     3061 ctaatctcag ctcgttacct gtataaaaga cacctgggag ccagaaatct ttctgattga
     3121 gagggggtca aatacttatt tccctcatta aaatgcaaat caatttataa catttttgac
     3181 atgcgttttt ctggattttt gttgttgtta ttctgtctct cactgttcaa ataaacctac
     3241 cattaaaatt atagactgat catttctttg tcagtgggca aacgtacaaa atcagcaggg
     3301 gatcaaatac ttttttccct cactgtaaag cttagagacg caagacactg cggatcgaga
     3361 cgcattgcgt ggttcgactg gccgtcgttt tacaacgaaa gggcctcgtg atacgcctat
     3421 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3481 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3541 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3601 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3661 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3721 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3781 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3841 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3901 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3961 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4021 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4081 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4141 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4201 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4261 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4321 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4381 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4441 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4501 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4561 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4621 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4681 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4741 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4801 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4861 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4921 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4981 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5041 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5101 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5161 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5221 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5281 ctgcgttatc ccctgattct gtgggtaacc gtattaccgc ctttgagtga gctgataccg
     5341 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gcacacagga
     5401 aacagctatg accgtctcgg ttggcagtga ctccgtctct gggccctaca gtgagggaaa
     5461 aaagtatttg atcccctgct gattttgtac gtttgcccac tgacaaagaa atgatcagtc
     5521 tataatttta atggtaggtt tatttgaaca gtgagagaca gaataacaac aacaaaatcc
     5581 agaaaaacgc atgtcaaaaa tgttataaat tgatttgcat tttaatgagg gaaataagta
     5641 tttgacccct ctgcaaaaca tgacctcgag ggccaggaag cttccctttg gtctccccct
     5701 cttaagtacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     5761 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     5821 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5881 ttctagttgt ggtttgtcca aactcaatcg atgcgagggg gcagggcctg catgtgaagg
     5941 gcgtcgtagg tgtccttggt ggctgtactg agaccctggt aaaggccatc gtgccccttg
     6001 cccctccggc gctcgccttt catcccaatc tcactgtagg cctccgccat cttatctttc
     6061 tgcagttcat tgtacaggcc ttcctgaggg ttcttccttc tcggctttcc ccccatctca
     6121 gggtcccggc cacgtctctt gtccaaaaca tcgtactcct ctcttcgtcc tagattgagc
     6181 tcgttataga gctggttctg gccctgctgg tacgcggggg cgtctgcgct cctgctgaac
     6241 ttcactctgg agcgataggc tgcgaagtcg cgtggtgggg catagggctg gtaatgcttg
     6301 cgggtgggcc cggggcggcg gggagtcatg ttcatgtagt cactgtgcag gagcctgctc
     6361 ctcttactcc tcacccagaa aataataaag gccactgtta ctagcaagct atagcaagcc
     6421 aggactcccc caaccaccac cagcacccaa aaatcacagg cgaagtccag ccccctcgtg
     6481 tgcactgcgc cccccgccgc tggccggcac gcctctgggc gcagggacag gggctgcgac
     6541 gcgatggtgg gcgccggtgt tggtggtcgc ggcgctggcg tcgtggtcag cactgtcagt
     6601 tgcgtgcctc caccgaaaac agcggctgaa ctatgccaaa tcatacagta ataatcggcc
     6661 tcgtcctcgg acctgagacc tgagatgagc agcacacccg cattggcgct tgcgtcttta
     6721 gatccactaa accttgatgg aactccagat ccctgctgct tatctgagtc gctcttgtaa
     6781 ttgagaaggt attggggagg agaacctggt ttttgctgat accagtagat cctgtaaggc
     6841 cctacattga ttcctgacct cagtgtacac gtcaaagaag cggaggcacc ggggcttgca
     6901 ctgagggagg atgattgcgt gagaacaggt tggctacccc cccctccgga cccgccccca
     6961 ccggaacctc caccgcctga gcccaggatc ccactggaca cggtgacggt tgtaccttgt
     7021 ccccaaacgt ccataccata gtagtaagtc atcatacccc gcgcacaata gtaaacagct
     7081 gtgtcttcgg gagttacact attaagctga agagagaact ggtttttgga agtatctgga
     7141 tttatagaca tcctgctttt cacacttaca gcataatcat tataccattt actgcggtag
     7201 tatgttctac cgagccattc caaccctcga ctgggggact gcctgatcca attccatgtc
     7261 gcggagttac tactcactga atcacctgat atggcacagg taagactaag agtctgggaa
     7321 ggagtgacca gaccggggcc agattgttgg agttgcactt gcggcctggc ggcgtggagc
     7381 agcaaggcca gcggcaggag caaggcggtc actggtaagg ccttatcgtc atcgtctttg
     7441 taatccatgg tggcgaccgg tggatccctc acgacacctg aaatggaaga aaaaaacttt
     7501 gaaccactgt ctgaggcttg agaatgaacc aagatccaaa ctcaaaaagg gcaaattcca
     7561 aggagaatta catcaagtgc caagctggcc taacttcagt ctccacccac tcagtgtggg
     7621 gaaactccat cgcataaaac ccctcccccc aacctaaaga cgacgtactc caaaagctcg
     7681 agaactaatc gaggtgcctg gacggcgccc ggtactcagt ggagtcacat gaagcgacgg
     7741 ctgaggacgg aaaggccctt ttcctttgtg tgggtgactc acccgcccgc tctcccgagc
     7801 gccgcgtcct ccattttgag ctccctgcag cagggccggg aagcggccat ctttccgctc
     7861 acgcaactgg tgccgaccgg gccagccttg ccgcccaggg cggggcgata
//
