LOCUS       sequence-442067-        4920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-3_FR Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4920)
  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 4920)
  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-Tc1-3_FR
FEATURES             Location/Qualifiers
     source          1..4920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      12..39
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(51..638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(131..150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(660..689)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     primer_bind     724..742
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     751..768
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     752..768
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1197..1696
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1232..1255)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1629..1648
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1710..1719
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1716..2315
                     /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     2215..2235
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2322..2378
                     /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(2379..2381,2382..2384,2385..3098)
                     /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(2379..2381,2382..2384,2385..3095)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2424..2445)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2685..2704)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3032..3053
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3120..3137)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3126..3350
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3624..3640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3624..3640)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3652..3669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      3721..3752
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3753..3855
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3856..3924,3925..4716)
                     /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(4074..4093)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4717..4908
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tgtaatcacc tggctcacct tcgggtgggc ctttctgcgt tgctggcgtt tttccatagg
       61 ctccgccccc ctgacgagca tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg
      121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      481 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac ctcggaaaaa
      541 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg attttctacc
      661 gaagaaaggc ccacccgtga aggtgagcca gtgagttgat tgcagtccag ttacgctgga
      721 gtcaataggc gtatcacgag gccctttcgt tgtaaaacga cggccagtcg aaccacgcaa
      781 tgcgtctcga tccgcagtgt cttgcgtctc tggtacctac agtgagagta aaaagtattt
      841 gatcccttgc tgattttgtt ggtttgtcca ctaataaaga catgatcatt ctatactttt
      901 aatggtagat gtattctaac atggagagac agaatatcaa aaagaaaatc aagaaaataa
      961 ctttaaagaa tttattttaa tttatttgtt tttcattgag ggaaataagt atttgatccc
     1021 ctagtatgca ttaggagttc tggctttcac agaccagtta gacgctccca aacaacttgt
     1081 tacctgaatt gaagacacca ggtctaacta atcacctgta tgtaagacaa ctgttcacag
     1141 aatcagacaa tcagacagat tccaaactct ccaccaccgg tgctagcaat tctaccgggt
     1201 aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac
     1261 ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca
     1321 accggctccg ttctttggtg gccccttcgc gccaccttct actccccccc tagtcaggaa
     1381 gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaagggaag tagcacgtct
     1441 cactagtctc gtgcagaggg acagcaccgc tgagcaatgg aggcgggtag gcttttgggg
     1501 cggcggccaa tagcagcttt gctccttcgc tttctgggct caggggcggg gagggggggg
     1561 gccggggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg
     1621 aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc
     1681 atctccgggc ctttcgacct gcagccaacg ccaccatggg gaccgagtac aagcccacgg
     1741 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg
     1801 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg
     1861 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     1921 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     1981 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     2041 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     2101 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     2161 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     2221 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     2281 cgcgcacctg gtgcatgacc cgcaagcccg gtgccatcga tgccactaac ttctccctgt
     2341 tgaaacaagc aggggatgtc gaagagaatc ccgggccaat ggtgagcaag ggcgaggagc
     2401 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2461 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     2521 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     2581 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2641 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2701 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2761 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2821 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2881 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2941 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     3001 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     3061 ccgggatcac tctcggcatg gacgagctgt acaagtaatt aattaagctc gctgatcagc
     3121 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3181 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3241 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3301 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3361 gcgccctcga ggtgcttttg gatagaggga tcaaatactt attaccctca atgaaaaaca
     3421 aataaattaa aataaattct ttaaagttat tttctggatt ttctttttga tattctgtct
     3481 ctccatgtta gaatacatct accattaaaa gtatagaatg atcatgtctt tattagtgga
     3541 caaaccaaca aaatcagcaa gggatcaaat actttttact ctcactgtag ggcccagaga
     3601 cggagtcact gccaaccgag acggtcatag ctgtttcctg tgtgccgctt cctcgctcac
     3661 tgactcgctg cgctcggtcg ttactgacca tttaaatcat acctgacctc catagcagaa
     3721 agtcaaaagc ctccgaccgg aggcttttga cttgatcggc acgtaagagg ttccaacttt
     3781 caccataatg aaataagatc actaccgggc gtattttttg agttatcgag attttcagga
     3841 gctaaggaag ctaaaatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3901 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3961 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4021 gagagtttac gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4081 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4141 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatctcac ggatggcatg
     4201 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4261 cttctggcaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4321 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4381 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4441 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4501 ggatcacttc tgcgctcggc cctcccggct ggctggttta ttgctgataa atctggagcc
     4561 ggtgagcgtg ggtcccgcgg tatcattgca gaactggggc cagatggtaa gccctcccgc
     4621 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4681 gctgagatag gtgcctcact gattaagcat tggtaagtga ccaaacagga aaaaaccgcc
     4741 cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag
     4801 ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt
     4861 taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgatg agggcccaaa
//
