LOCUS       sequence-441631-        6045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of hAT-3_AMi Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6045)
  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 6045)
  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-3_AMi
FEATURES             Location/Qualifiers
     source          1..6045
                     /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        1395..1894
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1430..1453)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1827..1846
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1908..1917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1914..2513
                     /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     2413..2433
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2520..2576
                     /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(2577..2579,2580..2582,2583..3296)
                     /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(2577..2579,2580..2582,2583..3293)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2622..2643)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2883..2902)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3230..3251
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3318..3335)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3324..3548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4148..4164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4148..4164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4176..4193)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4347..4935)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4427..4446)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5106..5897,5898..5966))
                     /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     5729..5748
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5967..6045,1..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 tcaattggct gcaggcagtg gcggatttac catgaaacaa accgtgcggt ggcacggggc
      241 ccccaatgac agggggcccc aaaatgcagg acaaatctga caaatgactg aacttaataa
      301 gcgttgcaag tcgcgaagcg gtccaggctt tctgcattct gcgaacttct tctgcgaact
      361 gtgctctaaa cgtggttttt tgaagtagtc tagcaaagaa tttgcttcgc cattttttcg
      421 ctgtctcggc aatagcataa cagcaactaa gggaaatcgt catttgccaa tcaaagtctg
      481 tctgcttgac caattatcca attgaacaaa agcaccctcc cccgcacttt gcaacaatgc
      541 ccgtgatttg tcactagtat gcaatggttc aaaatcccgt ttttacagag cctgataaaa
      601 acgcgggatt aactgcgtta aacctaattg ggtggcccac ccaggcccac ctgtggctac
      661 gcccctgctc tttcagacat ttttaatctg cctaaaatca aggtcagttt tttagcaata
      721 atgaagcgac actatgagtc tggcagtcaa aaatgcaaaa aaatgaaggc gaggcaagaa
      781 gatgaagcta agcaaaaaga cagtttgtca tcttatttta aaaagcttga aatattgaat
      841 agtggacatt catcaaaaag tatgggcgag gaaacatctg aattatctga tcacttgctg
      901 agttgtgggg ataccacaac gactgcaaaa gttgaacaag tggactctga agtgacaatg
      961 gaactacaaa atattggtaa ttttgtcgaa gtaagggaag aaacctaccc tgaagacatt
     1021 gcattatggc caaaatctgt ttcattggat atgatagaat atttcttagt aaataaaccg
     1081 aaaaacgtag gtaatatgga aaatttgaga aaaagagtat gaggttggag gacgaaagca
     1141 atttagaggt cttcatgagt cccattttct gaggatgaag aaaaatggga tgacagaaat
     1201 gagaatttgg ttgatttttt tcagaaacca ctaaggcagt ctactgttat gtttgcaaat
     1261 tattccctga ccatagtaaa ggaaaacaaa cattcgtcga cgggcactct aattggagaa
     1321 acattgcaag agatattgct gatcatgaaa cttccaaagc tcatattaat gctaccggtg
     1381 ctagcaattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt
     1441 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac
     1501 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac
     1561 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca
     1621 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
     1681 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
     1741 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg
     1801 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
     1861 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agccaacgcc accatgggga
     1921 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     1981 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     2041 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     2101 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     2161 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     2221 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     2281 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     2341 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     2401 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     2461 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gccatcgatg
     2521 ccactaactt ctccctgttg aaacaagcag gggatgtcga agagaatccc gggccaatgg
     2581 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2641 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2701 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2761 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     2821 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2881 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2941 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     3001 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     3061 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3121 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3181 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3241 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaattaa
     3301 ttaagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3361 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3421 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3481 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3541 ctctatggct tctgaggcgc gccctcgaga taaatatatt ttttcaaatc aaataagttt
     3601 cttcagagga agttaacttt tatttttcca ttcatgcatc atctatactt tttattttta
     3661 cacatttttc atgttagcgt aatgcaaata caagctttca tctagaccag aagttctcaa
     3721 actgtggtcc gagaactcca ttcaggtgga ccgaggaaag gttattaagg ttttttttta
     3781 taaagcgctg ttatccaaag cactttacaa tagttagcta acggtacaaa caacatttgg
     3841 aaagatcatt aagtggtccg ccgagaccct cagcaatttt caagtggtct gtggaaaaaa
     3901 aagttagact acaaaaaaat caaggtacct cactttattt tcatttactt cctattactt
     3961 ataatatagg aggaggatga agaaaaaaag aaatgaaaaa tgggggggag ataagagaga
     4021 atgttctttt tcttggctgg gtcccgaggg ggggggcgaa aatgaagctg cgcacagggc
     4081 cccactaact ctaaatccgc cactggctgc agggggccca gagacggagt cactgccaac
     4141 cgagacggtc atagctgttt cctgtgtgcc gcttcctcgc tcactgactc gctgcgctcg
     4201 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttacccaca
     4261 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4321 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4381 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4441 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4501 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4561 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4621 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4681 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4741 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4801 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4861 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4921 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4981 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5041 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5101 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct ctttcgttca
     5161 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5221 ggccccagtg ctgcaataat accgcgggac ccacgctcac cggctccaga tttatcagca
     5281 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5341 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5401 cgcaacgttg ttgccatcgc tacaggcatc gtggtatcac gctcgtcgtt tggtatggct
     5461 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgcgcaaa
     5521 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgccgtgtta
     5581 tcactcatgg ttatggcagc actacataat tctcttactg tcatgccatc cgtaagatgc
     5641 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5701 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5761 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5821 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5881 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5941 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6001 cagggttatt gtctcatgag cggatacata tttgaatgta tttag
//
