LOCUS       sequence-442080-        5317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Mariner2_AG Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5317)
  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 5317)
  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: pQLL-Mariner2_AG
FEATURES             Location/Qualifiers
     source          1..5317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(140..728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(220..239)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(899..1690,1691..1759))
                     /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     1522..1541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1760..1864)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1932..1950
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     1959..1976
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1960..1976
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2672..3171
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2707..2730)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3104..3123
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      3185..3194
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3191..3790
                     /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     3690..3710
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3797..3853
                     /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(3854..3856,3857..3859,3860..4573)
                     /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(3854..3856,3857..3859,3860..4570)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3899..3920)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4160..4179)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4507..4528
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4595..4612)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4601..4825
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5258..5274)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5258..5274)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5286..5303)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttaccc acagaatcag
       61 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      121 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      181 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      241 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      301 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      361 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      421 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      481 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      541 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      601 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      661 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      721 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      781 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      841 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      901 accaatgctt aatcagtgag gcacctatct cagcgatctg tctctttcgt tcatccatag
      961 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1021 gtgctgcaat aataccgcgg gacccacgct caccggctcc agatttatca gcaataaacc
     1081 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1141 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1201 ttgttgccat cgctacaggc atcgtggtat cacgctcgtc gtttggtatg gcttcattca
     1261 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgcgc aaaaaagcgg
     1321 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgccgtg ttatcactca
     1381 tggttatggc agcactacat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1441 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1501 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1561 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1621 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1681 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1741 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1801 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1861 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     1921 taacctataa aaataggcgt atcacgaggc cctttcgttg taaaacgacg gccagtcgaa
     1981 ccacgcaatg cgtctcgatc cgcagtgtct tgcgtctctc aattgtatac agtgtcggac
     2041 aaatcaatag gaccacttat caatttatta ttcacttatc atatttcatg acttaaagtg
     2101 aatcaatctt taccaaactt tcatgaaagc actattagaa tgtttacttt aagaatgttt
     2161 ttaaagtttg gtgaaaagtt aagaaacaaa aaagttatga taagaaatgt aaaaataacc
     2221 caaaaatcat gcgccaaaat aataggacca gttcagaaaa ttttaaaata attatcattt
     2281 attattgtta aaacacagtt tttgatgtta ttgagttgtc ggataactta acttaaatta
     2341 gtgaactaat gcgtattcat tttttttgta gaaacgctta gcgcagtggc aattcgctgt
     2401 gtctttaatt aacatgctct aatttactaa tgcaagaata ttacggcact cgcttaaggg
     2461 ttttgagctc cctttgcact gagagacgaa atagtggtga gtaccttcgt ctatgggtat
     2521 aaaagcgcgt cacagcgctg gatcgcttac actgtgttct aagccgttgc ctagaacact
     2581 tcacacgatc cattagctcg tgtatttttg ggtgcactac ttcaagcgct tgattgaagg
     2641 aattccaacg accggtgcta gcaattctac cgggtagggg aggcgctttt cccaaggcag
     2701 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
     2761 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
     2821 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
     2881 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc
     2941 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
     3001 ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg
     3061 ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa
     3121 aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc
     3181 caacgccacc atggggaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     3241 cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     3301 cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     3361 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     3421 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     3481 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     3541 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     3601 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     3661 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     3721 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     3781 gcccggtgcc atcgatgcca ctaacttctc cctgttgaaa caagcagggg atgtcgaaga
     3841 gaatcccggg ccaatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     3901 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     3961 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     4021 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     4081 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     4141 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     4201 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     4261 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     4321 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     4381 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     4441 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     4501 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     4561 gctgtacaag taattaatta agctcgctga tcagcctcga ctgtgccttc tagttgccag
     4621 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     4681 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     4741 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     4801 gctggggatg cggtgggctc tatggcttct gaggcgcgcc ctcgagatgc taccaagtta
     4861 gcccaggata aggctgtttt gttattttct ttagtttttc aaatgtacgg gctggccaca
     4921 ttgcgaatta aaagccggag ctgtaattat ttataggcaa taatgataaa tggttaaaat
     4981 aataaaattt tctatactgg tcctattatt ttgtcgcatg atttttgggg ttttttcaac
     5041 atttcttata ataacttttt tgtttgttaa cttatcaaca aactttcaaa gcagacttta
     5101 agccaacatc cgaaaagtgc tttcatgaaa atttggtaaa tattgattca ctttaggtaa
     5161 tgaaatgtga taagtgaata aaacattatt aagtggtcct attgatttgt ccgacactgt
     5221 atagggccca gagacggagt cactgccaac cgagacggtc atagctgttt cctgtgtgcc
     5281 gcttcctcgc tcactgactc gctgcgctcg gtcgttc
//
