LOCUS       sequence-442031-        5412 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Donor plasmid of HOBO Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5412)
  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 5412)
  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-HOBO
FEATURES             Location/Qualifiers
     source          1..5412
                     /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        532..1031
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(567..590)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     964..983
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1045..1054
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1051..1650
                     /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     1550..1570
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1657..1713
                     /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(1714..1716,1717..1719,1720..2433)
                     /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(1714..1716,1717..1719,1720..2430)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1759..1780)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2020..2039)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2367..2388
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2455..2472)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2461..2685
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3515..3531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3515..3531)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3543..3560)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3714..4302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3794..3813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4473..5264,5265..5333))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHFSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLRNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERKRQIAEIGAS
                     LIKHW"
     primer_bind     5096..5115
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5334..5412,1..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 tcatatggtt cacaccagag aactgcaagg gtggcacttt tttaccactc gactcacacc
      241 ctacaatttt gtgtgcgggt gctactcgcc acgcacatcg cgggtactta caaacacaca
      301 gtataaatct gaacatgcag acaagacacc ccgttgtgtg cgcacccgaa tcaatacggt
      361 gttttgcgtc gcgggtgccg ctcacacagt gcctaaaaag ggatgagtga gaaaaacact
      421 tgtgggtata ccgttaaaca catgggtgtt tccaaaaata ctcgggtgtt tccaaaaata
      481 ctcgagtggt ctcgtaggta gtcgagtcaa accggtgcta gcaattctac cgggtagggg
      541 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc
      601 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg
      661 ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc
      721 cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta
      781 gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg
      841 gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg
      901 ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc
      961 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc
     1021 cgggcctttc gacctgcagc caacgccacc atggggaccg agtacaagcc cacggtgcgc
     1081 ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac
     1141 taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg
     1201 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     1261 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     1321 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     1381 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     1441 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     1501 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     1561 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     1621 acctggtgca tgacccgcaa gcccggtgcc atcgatgcca ctaacttctc cctgttgaaa
     1681 caagcagggg atgtcgaaga gaatcccggg ccaatggtga gcaagggcga ggagctgttc
     1741 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1801 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1861 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1921 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1981 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2041 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2101 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2161 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     2221 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     2281 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     2341 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2401 atcactctcg gcatggacga gctgtacaag taattaatta agctcgctga tcagcctcga
     2461 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2521 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2581 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2641 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcgcgcc
     2701 ctcgagcagg taatataata acagaaaagc gaaatagatt atgcccaaaa tctgtagata
     2761 gcctcctttt tttgcattcc tattacaaaa acctaaacaa ctcgcaatag atattccttc
     2821 ttataagtat attttatatt attaattctt attatttgct taatttttgt ataagtgtta
     2881 agtattaagt ataagtatta attaataata tataagattg ttatttgtta agacattaga
     2941 tgcaaaatcc taaaaatgtg aaagtaatga agttccttat atttaataga tactttttaa
     3001 gcccactatg tttttattat ttagattgag acattaaaaa acgtaaaaat caacaaatgc
     3061 cgtctttaat tgcaattact ttatgtgttt gaaatgggag gcacccattg agtccatcaa
     3121 agagcaaaga catgagcaca aaaattttct tgggtattcc cttttaccct tcatttctta
     3181 tacccgtcac gcttccaccc atacaaattt taggcgtaca aaaaatgaga actgcagccc
     3241 gcatacaaaa aatgacctgc ggccgatcgt tgactgtgcg tccactcacc catacggctc
     3301 ttgcgcagca ggcctcgggt ggttttttta ctcgtaacaa aaacacaacg tcggtaaaac
     3361 actcgagtat tttgtgttgc cgcaagtagg gtgtcaaaaa aaacggggtg cctagagtac
     3421 cgagtgttta tcgggtggac gtagagtgcg agtggcgggc tgcagttctc tggttcacac
     3481 gggcccagag acggagtcac tgccaaccga gacggtcata gctgtttcct gtgtgccgct
     3541 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3601 tcaaaggcgg taatacggtt acccacagaa tcaggggata acgcaggaaa gaacatgtga
     3661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4141 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4381 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4441 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4501 atctcagcga tctgtctctt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4561 actacgatac gggagggctt accatctggc cccagtgctg caataatacc gcgggaccca
     4621 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4681 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4741 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccatcgctac aggcatcgtg
     4801 gtatcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4861 gttacatgat cccccatgtt gcgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4921 gtcagaagta agttggccgc cgtgttatca ctcatggtta tggcagcact acataattct
     4981 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5041 ttctgagaaa agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5101 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5161 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5221 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5281 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5401 gaatgtattt ag
//
