LOCUS       sequence-442186-        4780 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Donor plasmid of hAT-6_PM Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4780)
  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 4780)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-6_PM
FEATURES             Location/Qualifiers
     source          1..4780
                     /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        502..1001
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(537..560)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     934..953
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1015..1024
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1021..1620
                     /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     1520..1540
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1627..1683
                     /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(1684..1686,1687..1689,1690..2403)
                     /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(1684..1686,1687..1689,1690..2400)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1729..1750)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1990..2009)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2337..2358
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2425..2442)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2431..2655
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2883..2899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2883..2899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2911..2928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3082..3670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3162..3181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3841..4632,4633..4701))
                     /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     4464..4483
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4702..4780,1..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 taagctttta gagcacaggg gtcgggaacc tatggctcgc gagccagatg tggctctttt
      241 gatgattgca tatggctcgc agataaaaca aaatattctc acttgtttta atccatccat
      301 cgatttttca ctgcgcatgt cctgttcagg gctgcaggag caattgtcca ttattttaat
      361 tggatgatac tggcctacgt ttgccgttgc tgggtaaaac aggtaaacgc ccccttttga
      421 atcagtctga gactgcgtct gctcggtcat tagctcaaag ctaacccttc gacgaagaag
      481 accggtgcta gcaattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
      541 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
      601 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
      661 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
      721 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
      781 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
      841 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
      901 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
      961 ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc caacgccacc
     1021 atggggaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc
     1081 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg
     1141 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     1201 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     1261 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     1321 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     1381 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     1441 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     1501 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     1561 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     1621 atcgatgcca ctaacttctc cctgttgaaa caagcagggg atgtcgaaga gaatcccggg
     1681 ccaatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1741 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1801 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1861 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     1921 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1981 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2041 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2101 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     2161 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     2221 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2281 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     2341 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     2401 taattaatta agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2461 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2521 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2581 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2641 cggtgggctc tatggcttct gaggcgcgcc ctcgagtggt gagtgttata acaacattat
     2701 ttaaaagaat aaattcagag gcttattata ctgacattta gttgaattca cttttaaaat
     2761 atattatatg gctctcactg aaataaattt ccaaattttt tgctttcatg gctctcttag
     2821 tcaaaaaggt tcccgacccc tgttagaggg gcccagagac ggagtcactg ccaaccgaga
     2881 cggtcatagc tgtttcctgt gtgccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2941 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttac ccacagaatc
     3001 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3061 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3121 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3181 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3241 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3301 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3361 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3421 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3481 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3541 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3601 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3661 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3721 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3781 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3841 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctctttc gttcatccat
     3901 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3961 cagtgctgca ataataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa
     4021 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4081 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4141 cgttgttgcc atcgctacag gcatcgtggt atcacgctcg tcgtttggta tggcttcatt
     4201 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgc gcaaaaaagc
     4261 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgccg tgttatcact
     4321 catggttatg gcagcactac ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4381 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4441 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4501 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4561 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4621 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4681 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4741 ttattgtctc atgagcggat acatatttga atgtatttag
//
