LOCUS       sequence-445643-        8068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of SB100X Transposon with 5 kb cargo gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8068)
  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 8068)
  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-SB100X-5 kb
FEATURES             Location/Qualifiers
     source          1..8068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     507..527
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      574..583
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             583..606
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2149..2230
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2174..2193
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2765..2781)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2765..2781)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2793..2810)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2964..3552)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3044..3063)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3723..4514,4515..4583))
                     /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     4346..4365
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4584..4688)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4756..4774
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4783..4800
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4784..4800
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5250..5749
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5285..5308)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5682..5701
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      5763..5772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5769..6368
                     /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     6268..6288
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             6375..6431
                     /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(6432..6434,6435..6437,6438..7151)
                     /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(6432..6434,6435..6437,6438..7148)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6477..6498)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6738..6757)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7085..7106
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7173..7190)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7179..7403
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        join(7449..8068,1..559)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(7680..8068,1..550)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
ORIGIN
        1 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
       61 acgggggtag tctcaagctc gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
      121 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
      181 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
      241 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
      301 tgactccact gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
      361 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
      421 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
      481 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
      541 tccatttcag gtgtcgtgag ggatccaccg gtcgccacca tggattacaa agacgatgac
      601 gataaggcct taccagtgac cgccttgctc ctgccgctgg ccttgctgct ccacgccgcc
      661 aggccgcaag tgcaactcca acaatctggc cccggtctgg tcactccttc ccagactctt
      721 agtcttacct gtgccatatc aggtgattca gtgagtagta actccgcgac atggaattgg
      781 atcaggcagt cccccagtcg agggttggaa tggctcggta gaacatacta ccgcagtaaa
      841 tggtataatg attatgctgt aagtgtgaaa agcaggatgt ctataaatcc agatacttcc
      901 aaaaaccagt tctctcttca gcttaatagt gtaactcccg aagacacagc tgtttactat
      961 tgtgcgcggg gtatgatgac ttactactat ggtatggacg tttggggaca aggtacaacc
     1021 gtcaccgtgt ccagtgggat cctgggctca ggcggtggag gttccggtgg gggcgggtcc
     1081 ggaggggggg gtagccaacc tgttctcacg caatcatcct ccctcagtgc aagccccggt
     1141 gcctccgctt ctttgacgtg tacactgagg tcaggaatca atgtagggcc ttacaggatc
     1201 tactggtatc agcaaaaacc aggttctcct ccccaatacc ttctcaatta caagagcgac
     1261 tcagataagc agcagggatc tggagttcca tcaaggttta gtggatctaa agacgcaagc
     1321 gccaatgcgg gtgtgctgct catctcaggt ctcaggtccg aggacgaggc cgattattac
     1381 tgtatgattt ggcatagttc agccgctgtt ttcggtggag gcacgcaact gacagtgctg
     1441 accacgacgc cagcgccgcg accaccaaca ccggcgccca ccatcgcgtc gcagcccctg
     1501 tccctgcgcc cagaggcgtg ccggccagcg gcggggggcg cagtgcacac gagggggctg
     1561 gacttcgcct gtgatttttg ggtgctggtg gtggttgggg gagtcctggc ttgctatagc
     1621 ttgctagtaa cagtggcctt tattattttc tgggtgagga gtaagaggag caggctcctg
     1681 cacagtgact acatgaacat gactccccgc cgccccgggc ccacccgcaa gcattaccag
     1741 ccctatgccc caccacgcga cttcgcagcc tatcgctcca gagtgaagtt cagcaggagc
     1801 gcagacgccc ccgcgtacca gcagggccag aaccagctct ataacgagct caatctagga
     1861 cgaagagagg agtacgatgt tttggacaag agacgtggcc gggaccctga gatgggggga
     1921 aagccgagaa ggaagaaccc tcaggaaggc ctgtacaatg aactgcagaa agataagatg
     1981 gcggaggcct acagtgagat tgggatgaaa ggcgagcgcc ggaggggcaa ggggcacgat
     2041 ggcctttacc agggtctcag tacagccacc aaggacacct acgacgccct tcacatgcag
     2101 gccctgcccc ctcgcatcga ttgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2161 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2221 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     2281 ggaggttttt taaagcaagt aaaacctcta caaatgtggt acttaagagg gggagaccaa
     2341 agggaagctt cctggccctc gagccttgaa atacatccac aggtacacct ccaattgact
     2401 caaatgatgt caattagtct atcagaagct tgtggaaggc tactcgaaat gtttgaccca
     2461 agttaaacaa tttaaaggca atgctaccaa atactaattg agtgtatgta aacttctgac
     2521 ccactgggaa tgtgatgaaa gaaataaaag ctgaaatgaa tcattctctc tactattatt
     2581 ctgatatttc acattcttaa aataaagtgg tgatcctaac tgacctaaga cagggaattt
     2641 ttactaggat taaatgtcag gaattgtgaa aaagtgagtt taaatgtatt tggctaaggt
     2701 gtatgtaaac ttccgacttc aactgtatag gggcccagag acggagtcac tgccaaccga
     2761 gacggtcata gctgtttcct gtgtgccgct tcctcgctca ctgactcgct gcgctcggtc
     2821 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt acccacagaa
     2881 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2941 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3001 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3061 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3121 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3181 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3241 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3301 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3361 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3421 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3481 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3541 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3601 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3661 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3721 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctctt tcgttcatcc
     3781 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3841 cccagtgctg caataatacc gcgggaccca cgctcaccgg ctccagattt atcagcaata
     3901 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3961 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4021 aacgttgttg ccatcgctac aggcatcgtg gtatcacgct cgtcgtttgg tatggcttca
     4081 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gcgcaaaaaa
     4141 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc cgtgttatca
     4201 ctcatggtta tggcagcact acataattct cttactgtca tgccatccgt aagatgcttt
     4261 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4321 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4381 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4441 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4501 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4561 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4621 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4681 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     4741 acattaacct ataaaaatag gcgtatcacg aggccctttc gttgtaaaac gacggccagt
     4801 cgaaccacgc aatgcgtctc gatccgcagt gtcttgcgtc tctactagtc tatacagttg
     4861 aagtcggaag tttacataca cttaagttgg agtcattaaa actcgttttt caactactcc
     4921 acaaatttct tgttaacaaa caatagtttt ggcaagtcag ttaggacatc tactttgtgc
     4981 atgacacaag tcatttttcc aacaattgtt tacagacaga ttatttcact tataattcac
     5041 tgtatcacaa ttccagtggg tcagaagttt acatacacta agttgactgt gcctttaaac
     5101 agcttggaaa attccagaaa atgatgtcat ggctttagaa gcttcccaca ataagttggg
     5161 tgaattttgg ctcattcctc ctttctatag gattgaggtc agagctttgt gatggaattc
     5221 tgtggaatac cggtgctagc aattctaccg ggtaggggag gcgcttttcc caaggcagtc
     5281 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
     5341 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
     5401 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
     5461 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
     5521 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
     5581 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
     5641 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
     5701 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagcca
     5761 acgccaccat ggggaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     5821 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     5881 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     5941 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     6001 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     6061 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     6121 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     6181 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     6241 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     6301 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     6361 ccggtgccat cgatgccact aacttctccc tgttgaaaca agcaggggat gtcgaagaga
     6421 atcccgggcc aatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     6481 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     6541 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     6601 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     6661 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     6721 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     6781 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     6841 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     6901 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     6961 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     7021 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     7081 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     7141 tgtacaagta attaattaag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     7201 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     7261 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     7321 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     7381 tggggatgcg gtgggctcta tggcttctga gggcggccgc caatgggtac cttggttaat
     7441 taacgtgagg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     7501 agttgggggg aggggtcggc aattgacccg gtgcctagag aaggtggcgc ggggtaaact
     7561 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     7621 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
     7681 taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc
     7741 cttgaattac ttccacctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa
     7801 gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg
     7861 aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc
     7921 tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt
     7981 ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt
     8041 ttggggccgc gggcggcgac ggggcccg
//
