LOCUS       sequence-443658-        4969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of TC1-2_DM Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4969)
  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 4969)
  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_DM
FEATURES             Location/Qualifiers
     source          1..4969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..20
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     29..46
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     30..46
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        474..973
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(509..532)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     906..925
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      987..996
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             993..1592
                     /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     1492..1512
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1599..1655
                     /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(1656..1658,1659..1661,1662..2375)
                     /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(1656..1658,1659..1661,1662..2372)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1701..1722)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1962..1981)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2309..2330
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2397..2414)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2403..2627
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2951..2967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2951..2967)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2979..2996)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      3048..3079
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3080..3182
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3183..3251,3252..4043)
                     /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(3401..3420)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4044..4235
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4259..4286
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(4298..4885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4378..4397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4907..4936)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 caataggcgt atcacgaggc cctttcgttg taaaacgacg gccagtcgaa ccacgcaatg
       61 cgtctcgatc cgcagtgtct tgcgtctcta agctttacac tagtgggcat aagtatttag
      121 acaccattag cttgcaactg ccgggctatg cttttcatag tacagtcaaa taaataagta
      181 aactattgat gataaaacaa agagtgcaat tacagttaca cgcgagaggt atgttttgcc
      241 gtttaccttt tcaattatcc ttctgctgat aagaaagcta gacagacaac gatttgggcg
      301 gtggcaaaag tatttagaca tcacatttaa ttagttcgca ttgatctgtt gttcgaagaa
      361 tatcaaattt ttgacttttt tagatcattt gaacatttgt tcttgataac gtgttactac
      421 ctacttttat ttcattcata aatttgttaa acaccggtgc tagcaattct accgggtagg
      481 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg
      541 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc
      601 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt
      661 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac
      721 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag
      781 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc
      841 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg
      901 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc
      961 tccgggcctt tcgacctgca gccaacgcca ccatggggac cgagtacaag cccacggtgc
     1021 gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg
     1081 actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc
     1141 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     1201 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     1261 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     1321 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     1381 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     1441 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     1501 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     1561 gcacctggtg catgacccgc aagcccggtg ccatcgatgc cactaacttc tccctgttga
     1621 aacaagcagg ggatgtcgaa gagaatcccg ggccaatggt gagcaagggc gaggagctgt
     1681 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     1741 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     1801 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     1861 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     1921 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     1981 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2041 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2101 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     2161 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2221 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     2281 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2341 ggatcactct cggcatggac gagctgtaca agtaattaat taagctcgct gatcagcctc
     2401 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2461 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2521 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2581 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgcg
     2641 ccctcgagtt tgtcataatt ttttagttga cttttttttt tcaatttata acatttaaat
     2701 aatttgtcca aatacttatg ccactgctaa tttgtaaaaa agtaattttt gttaaatcaa
     2761 caatgaagtt atccatatgt ttatgttacc taatttaact acgatagtct acctatttac
     2821 taaaattatt aaacaatttt gctttaagta aacaatgagt tacaaacaaa tatattgaat
     2881 atatttcttg tccaaatact tatgcccact agtgtagggc ccagagacgg agtcactgcc
     2941 aaccgagacg gtcatagctg tttcctgtgt gccgcttcct cgctcactga ctcgctgcgc
     3001 tcggtcgtta ctgaccattt aaatcatacc tgacctccat agcagaaagt caaaagcctc
     3061 cgaccggagg cttttgactt gatcggcacg taagaggttc caactttcac cataatgaaa
     3121 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     3181 aaatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3241 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3301 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agtttacgcc
     3361 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3421 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3481 tggttgagta ctcaccagtc acagaaaagc atctcacgga tggcatgaca gtaagagaat
     3541 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctggcaacga
     3601 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3661 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3721 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3781 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga tcacttctgc
     3841 gctcggccct cccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3901 cccgcggtat cattgcagaa ctggggccag atggtaagcc ctcccgcatc gtagttatct
     3961 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4021 cctcactgat taagcattgg taagtgacca aacaggaaaa aaccgccctt aacatggccc
     4081 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
     4141 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
     4201 tcgcgcgttt cggtgatgac ggtgaaaacc tctgatgagg gcccaaatgt aatcacctgg
     4261 ctcaccttcg ggtgggcctt tctgcgttgc tggcgttttt ccataggctc cgcccccctg
     4321 acgagcatca caaaaatcga tgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4381 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4441 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4501 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4561 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4621 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4681 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4741 cagtatttgg tatctgcgct ctgctgaagc cagttacctc ggaaaaagag ttggtagctc
     4801 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4861 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatt ttctaccgaa gaaaggccca
     4921 cccgtgaagg tgagccagtg agttgattgc agtccagtta cgctggagt
//
