LOCUS       sequence-443536-        4787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Mariner-4_AMi Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4787)
  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 4787)
  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-4_AMi
FEATURES             Location/Qualifiers
     source          1..4787
                     /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        298..797
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(333..356)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     730..749
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      811..820
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             817..1416
                     /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     1316..1336
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1423..1479
                     /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(1480..1482,1483..1485,1486..2199)
                     /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(1480..1482,1483..1485,1486..2196)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1525..1546)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1786..1805)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2133..2154
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2221..2238)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2227..2451
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2890..2906)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2890..2906)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2918..2935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3089..3677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3169..3188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3848..4639,4640..4708))
                     /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     4471..4490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4709..4787,1..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 taagctttac aggcgctcct cacttaacga cctacctgtt taacgaccgc gcggacttac
      241 gaccgcgcgg tcgtggaaac ggaagtactg tacaagaccg gtgctagcaa ttctaccggg
      301 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
      361 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
      421 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
      481 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
      541 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
      601 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
      661 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
      721 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
      781 catctccggg cctttcgacc tgcagccaac gccaccatgg ggaccgagta caagcccacg
      841 gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc
      901 gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc
      961 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     1021 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     1081 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     1141 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     1201 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     1261 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     1321 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     1381 ccgcgcacct ggtgcatgac ccgcaagccc ggtgccatcg atgccactaa cttctccctg
     1441 ttgaaacaag caggggatgt cgaagagaat cccgggccaa tggtgagcaa gggcgaggag
     1501 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1561 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1621 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1681 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1741 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1801 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1861 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1921 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1981 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2041 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     2101 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2161 gccgggatca ctctcggcat ggacgagctg tacaagtaat taattaagct cgctgatcag
     2221 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2281 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2341 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2401 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2461 cgcgccctcg agcgggattc ttctcgaaag ctacacccag gaatccacac tttgaaggat
     2521 tcgatgacta aaatgttaat ctgaatgtct ttttttataa ctttaatttt tttggtacat
     2581 catttttata attttcgctg attgtatttt tatgttctgt aatatgtaag agttatgcaa
     2641 attaaaacaa agttatcgag ttggagtttc ctttagacct tttctttatt cagaataaat
     2701 aaaaacgatt acctgtattt acagtatgca aacctaagta atgtatatta gagctgattt
     2761 cctctattca ggggcggctc ctcgcttatc gaccaattcg cttaacgacc tagtcgtggg
     2821 aacggaactc ggtcgttaag tgaggagtgc ctgtagggcc cagagacgga gtcactgcca
     2881 accgagacgg tcatagctgt ttcctgtgtg ccgcttcctc gctcactgac tcgctgcgct
     2941 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttaccca
     3001 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3061 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3121 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3181 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3241 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3301 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3361 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3421 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3481 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3541 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3601 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3661 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3721 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3781 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3841 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctctttcgtt
     3901 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3961 ctggccccag tgctgcaata ataccgcggg acccacgctc accggctcca gatttatcag
     4021 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4081 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4141 tgcgcaacgt tgttgccatc gctacaggca tcgtggtatc acgctcgtcg tttggtatgg
     4201 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgcgca
     4261 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgccgtgt
     4321 tatcactcat ggttatggca gcactacata attctcttac tgtcatgcca tccgtaagat
     4381 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4441 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4501 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4561 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4621 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4681 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4741 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttag
//
