LOCUS       sequence-441658-        4895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of hAT-12_Crp Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4895)
  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 4895)
  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-12_Crp
FEATURES             Location/Qualifiers
     source          1..4895
                     /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        282..781
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(317..340)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     714..733
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      795..804
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             801..1400
                     /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     1300..1320
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1407..1463
                     /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(1464..1466,1467..1469,1470..2183)
                     /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(1464..1466,1467..1469,1470..2180)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1509..1530)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1770..1789)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2117..2138
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2205..2222)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2211..2435
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2998..3014)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2998..3014)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3026..3043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3197..3785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3277..3296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3956..4747,4748..4816))
                     /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     4579..4598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4817..4895,1..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 tctgcagcta tatagtaggg ctgtgcgaag cttcagtagc tgattcgact ggaggagatt
      241 cggcccgatt cggcggccaa accggtgcta gcaattctac cgggtagggg aggcgctttt
      301 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
      361 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
      421 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
      481 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
      541 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
      601 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
      661 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
      721 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc
      781 gacctgcagc caacgccacc atggggaccg agtacaagcc cacggtgcgc ctcgccaccc
      841 gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
      901 cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct
      961 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     1021 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     1081 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     1141 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     1201 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     1261 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     1321 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     1381 tgacccgcaa gcccggtgcc atcgatgcca ctaacttctc cctgttgaaa caagcagggg
     1441 atgtcgaaga gaatcccggg ccaatggtga gcaagggcga ggagctgttc accggggtgg
     1501 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1561 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1621 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1681 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1741 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1801 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1861 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1921 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1981 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2041 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     2101 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2161 gcatggacga gctgtacaag taattaatta agctcgctga tcagcctcga ctgtgccttc
     2221 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2281 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2341 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2401 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcgcgcc ctcgaggtgc
     2461 caccctcctc actccgcacc tatttgcttt ttttatttgc tttcagaacc ttgaggtgca
     2521 ctttcacaga aacccctgca ccactctgcc tgaaacctgg caggcctcat gcccgcacga
     2581 ggggctacca tccctgctgt tttcatcccg ctctgcctta acacacacaa gtgatacgag
     2641 ttttgctttc aacagtttga ggtgcacttt cacagaaacc cctgcaccta tctccttgaa
     2701 acttggcagg cttcatgccc tcagaagggg ctaccatccc tgctgttttc atccgaatct
     2761 gcaaaaaaat gacaaagtta taggtatttt agtgattccc cattatagtc tatggccgaa
     2821 tctccgaatc ggcgccgaat ctccgaatcc gaatcgaatc gggacagtga ttcgaatcgc
     2881 cgaatcgaat cactgtccct ccgaatcggc cgaatccgaa tccgaagcga atacttgctg
     2941 cttcgcacag ccctactata taggggccca gagacggagt cactgccaac cgagacggtc
     3001 atagctgttt cctgtgtgcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3061 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttacccaca gaatcagggg
     3121 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3181 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3241 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3301 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3361 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3421 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3481 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3541 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3601 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3661 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3721 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3781 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3841 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3901 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3961 aatgcttaat cagtgaggca cctatctcag cgatctgtct ctttcgttca tccatagttg
     4021 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4081 ctgcaataat accgcgggac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4141 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4201 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4261 ttgccatcgc tacaggcatc gtggtatcac gctcgtcgtt tggtatggct tcattcagct
     4321 ccggttccca acgatcaagg cgagttacat gatcccccat gttgcgcaaa aaagcggtta
     4381 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgccgtgtta tcactcatgg
     4441 ttatggcagc actacataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4501 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4561 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4621 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4681 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4741 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4801 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4861 gtctcatgag cggatacata tttgaatgta tttag
//
