LOCUS       sequence-441632-        4775 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Donor plasmid of TC1_FR1 Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4775)
  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 4775)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_FR1
FEATURES             Location/Qualifiers
     source          1..4775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..112
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     121..138
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     122..138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        358..857
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(393..416)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     790..809
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      871..880
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             877..1476
                     /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     1376..1396
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1483..1539
                     /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(1540..1542,1543..1545,1546..2259)
                     /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(1540..1542,1543..1545,1546..2256)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1585..1606)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1846..1865)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2193..2214
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2281..2298)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2287..2511
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2878..2894)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2878..2894)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2906..2923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3077..3665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3157..3176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3836..4627,4628..4696))
                     /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     4459..4478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4697..4775,1..26))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
       61 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
      121 tgtaaaacga cggccagtcg aaccacgcaa tgcgtctcga tccgcagtgt cttgcgtctc
      181 tcaattgtac actcaccggc cactttatta ggtacacctg ttcaattgct tgttaacaca
      241 aatagctaat cagccaatca catggccgca actcaatgca tttaggcatg tagacgtggt
      301 caagacaact tgctgaagtt caaaccgagc atcagaaccg gtgctagcaa ttctaccggg
      361 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
      421 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
      481 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
      541 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
      601 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
      661 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
      721 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
      781 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
      841 catctccggg cctttcgacc tgcagccaac gccaccatgg ggaccgagta caagcccacg
      901 gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc
      961 gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc
     1021 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     1081 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     1141 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     1201 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     1261 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     1321 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     1381 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     1441 ccgcgcacct ggtgcatgac ccgcaagccc ggtgccatcg atgccactaa cttctccctg
     1501 ttgaaacaag caggggatgt cgaagagaat cccgggccaa tggtgagcaa gggcgaggag
     1561 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1621 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1681 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1741 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1801 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1861 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1921 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1981 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     2041 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2101 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     2161 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2221 gccgggatca ctctcggcat ggacgagctg tacaagtaat taattaagct cgctgatcag
     2281 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2341 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2401 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2461 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2521 cgcgccctcg agcatctcaa actggtttct ggaacatgac aatgagttca ctgtactcca
     2581 gtggcctcca cagtcaccag atctcaatcc aatagagcac ctttgggatg tggtggaacg
     2641 ggagattcgc atcatagatg tgcagccgac aaatctgcag caactgcgtg atgctatcat
     2701 gtcaatatgg accaaaatct ctgaggaatg tttccaacac cttgttgaaa gtatgccacg
     2761 aagaattaag gcagttctga aggcaaaagg gggtccaacc ttttactagc aaggtgtacc
     2821 taataaagtg gccggtgagt gtagggccca gagacggagt cactgccaac cgagacggtc
     2881 atagctgttt cctgtgtgcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2941 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttacccaca gaatcagggg
     3001 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3061 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3481 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3841 aatgcttaat cagtgaggca cctatctcag cgatctgtct ctttcgttca tccatagttg
     3901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3961 ctgcaataat accgcgggac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4141 ttgccatcgc tacaggcatc gtggtatcac gctcgtcgtt tggtatggct tcattcagct
     4201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgcgcaaa aaagcggtta
     4261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgccgtgtta tcactcatgg
     4321 ttatggcagc actacataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4741 gtctcatgag cggatacata tttgaatgta tttag
//
