LOCUS       sequence-443566-        4727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Mariner-3_Crp Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4727)
  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 4727)
  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-Mariner-3_Crp
FEATURES             Location/Qualifiers
     source          1..4727
                     /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        332..831
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(367..390)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     764..783
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      845..854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             851..1450
                     /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     1350..1370
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1457..1513
                     /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(1514..1516,1517..1519,1520..2233)
                     /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(1514..1516,1517..1519,1520..2230)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1559..1580)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1820..1839)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2167..2188
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2255..2272)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2261..2485
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2830..2846)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2830..2846)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2858..2875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3029..3617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3109..3128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3788..4579,4580..4648))
                     /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     4411..4430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4649..4727,1..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 agtagaaccc cgttttacgc gtattcgact tacccgagtc tcgcgtttac
      241 gcgaattaat attagcaccc aagcagaaaa atacaccccg agacgcgcgt cattcgcgta
      301 tatacgcgag accggtgcta gcaattctac cgggtagggg aggcgctttt cccaaggcag
      361 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
      421 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
      481 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
      541 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc
      601 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
      661 ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg
      721 ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa
      781 aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc
      841 caacgccacc atggggaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
      901 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
      961 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     1021 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     1081 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     1141 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     1201 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     1261 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     1321 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     1381 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     1441 gcccggtgcc atcgatgcca ctaacttctc cctgttgaaa caagcagggg atgtcgaaga
     1501 gaatcccggg ccaatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1561 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1621 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1681 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     1741 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1801 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1861 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1921 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1981 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2041 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2101 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2161 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2221 gctgtacaag taattaatta agctcgctga tcagcctcga ctgtgccttc tagttgccag
     2281 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2341 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2401 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2461 gctggggatg cggtgggctc tatggcttct gaggcgcgcc ctcgagtgga tggattgaag
     2521 attgttgcat gtccattttg tattttttta ttattttact tataatgtaa atgaataaat
     2581 atgtttacaa tacgtttttg taacataata aataaagtat tggttgaaaa aggttgtatt
     2641 tggtgccttt ggtgtcataa atgtgaggtc caaggaacgc aacccctcac ttttctatta
     2701 tttcctatgg gaaaattcac ctcgagttgc gcgaattcga catacgcgag gtttctcagg
     2761 aacgcaatat acgcgtaagt cggggttcta ctgtagggcc cagagacgga gtcactgcca
     2821 accgagacgg tcatagctgt ttcctgtgtg ccgcttcctc gctcactgac tcgctgcgct
     2881 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttaccca
     2941 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3001 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3061 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3121 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3181 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3241 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3301 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3361 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3421 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3481 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3541 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3601 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3661 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3721 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3781 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctctttcgtt
     3841 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3901 ctggccccag tgctgcaata ataccgcggg acccacgctc accggctcca gatttatcag
     3961 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4021 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4081 tgcgcaacgt tgttgccatc gctacaggca tcgtggtatc acgctcgtcg tttggtatgg
     4141 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgcgca
     4201 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgccgtgt
     4261 tatcactcat ggttatggca gcactacata attctcttac tgtcatgcca tccgtaagat
     4321 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4381 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4441 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4501 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4561 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4621 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4681 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttag
//
