LOCUS       sequence-441635-        5638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of hAT-8_AMi Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5638)
  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 5638)
  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-hAT-8_AMi
FEATURES             Location/Qualifiers
     source          1..5638
                     /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        1375..1874
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1410..1433)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1807..1826
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1888..1897
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1894..2493
                     /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     2393..2413
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2500..2556
                     /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(2557..2559,2560..2562,2563..3276)
                     /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(2557..2559,2560..2562,2563..3273)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2602..2623)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2863..2882)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3210..3231
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3298..3315)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3304..3528
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3741..3757)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3741..3757)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3769..3786)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3940..4528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4020..4039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4699..5490,5491..5559))
                     /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     5322..5341
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5560..5638,1..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 tcaattggtt gtaaccaagg ttccctctaa gctgcgtgcg tgcgcggctg cgcacgcttt
      241 aaaaccatgc tgcgcacgag ctcattagtg ccgtgcaccc acccggtgag gagctgggtt
      301 gcggcttcga gaggagcggg caagaagctg agcacggcag aggcaccgac gccggcgtgg
      361 gaaacttacc gaggtaagct caggcagcgg ctgctcccga ggccagtgct ccagaggcga
      421 ggtggggcag gcgggggctg gggctgggga actgctccgc ttcccagccc cggcgccatg
      481 tgcctccttg ctcccagagg catggcgagg tgggggctgg gggctgggcc aggaaactgc
      541 tccgctcccg gaggcaagcg acatggcgct aggctcccgg ccctgagggc caagaccgac
      601 cggctaggtg gtggctcggg gccggggccg ggaccgggaa actgctccgt cccagcagcg
      661 atgcaggggc tggggctggg ccgaggccag tggcggtgcg aggctggggc caggctactc
      721 ctgcactgcc aaatacgacg atgcgggtaa ggctggaaat ggccaggtat gattcggaat
      781 aagtttggga aatgcagcaa gttcccatga aatcagtgcc tacttatcaa gaggagctgt
      841 atgagtggga cttggcctct ggttcttact ttccaccatc aaaaacagca tgttggtaag
      901 tgaaaaataa aaataaatga attccaaatg aacacccaat cctctactcc ctaaaatgca
      961 ttcgctgcag aaatcaaagc agatttcaaa tcaaaagaaa ggccagtgtt tatggccaag
     1021 gttccctcca agctgcgctc ccggaggcaa ggcgggggct ggggctggcg ccagtttttt
     1081 agccatttag tggtttgtct ttattgtcac tagtttatgc atttgaattt aaaagtatta
     1141 aacagaaaaa gaagttgcaa atggtgttgt ttttgttaaa agcatcctaa ctgtgtaatc
     1201 ttgagtttgg gggtgctggt tacagctgtg ttggtctagg ggcataggca gacaaggtgc
     1261 tttgggtaaa ttagctgtta gctgcttttt ctatcattct aggaaggcca atatcttccc
     1321 tgcagcaagg atcatcttta tattcatttt acaaccggtg ctagcaattc taccgggtag
     1381 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
     1441 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
     1501 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
     1561 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
     1621 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
     1681 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
     1741 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
     1801 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
     1861 ctccgggcct ttcgacctgc agccaacgcc accatgggga ccgagtacaa gcccacggtg
     1921 cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc
     1981 gactaccccg ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag
     2041 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     2101 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     2161 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     2221 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     2281 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     2341 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     2401 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     2461 cgcacctggt gcatgacccg caagcccggt gccatcgatg ccactaactt ctccctgttg
     2521 aaacaagcag gggatgtcga agagaatccc gggccaatgg tgagcaaggg cgaggagctg
     2581 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2641 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2701 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     2761 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2821 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2881 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2941 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     3001 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     3061 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3121 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     3181 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3241 gggatcactc tcggcatgga cgagctgtac aagtaattaa ttaagctcgc tgatcagcct
     3301 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     3361 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3421 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3481 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgc
     3541 gccctcgaga taaaggttat agtatcatct tttgttgtta tactgtgggg cggaccgaag
     3601 cggcgcgcgc cgaaaggggg aggcgggacc aattcgggaa cggaccaagc ggctgaagcg
     3661 gcgcgctcac ggaccgctac accttagagg gaacattggt tgtaacgggc ccagagacgg
     3721 agtcactgcc aaccgagacg gtcatagctg tttcctgtgt gccgcttcct cgctcactga
     3781 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3841 acggttaccc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3901 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3961 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4021 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4081 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4141 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4201 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4261 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4321 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4381 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4441 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4501 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4561 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4621 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4681 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4741 tctctttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4801 gggcttacca tctggcccca gtgctgcaat aataccgcgg gacccacgct caccggctcc
     4861 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4921 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4981 agttaatagt ttgcgcaacg ttgttgccat cgctacaggc atcgtggtat cacgctcgtc
     5041 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5101 catgttgcgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5161 ggccgccgtg ttatcactca tggttatggc agcactacat aattctctta ctgtcatgcc
     5221 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5281 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5341 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5401 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5461 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5521 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5581 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttag
//
