LOCUS       sequence-315752-        4464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression construct with ubiquitous expression to test fluorescence
            level on microscope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4464)
  AUTHORS   Aljohani MD, El Mouridi S, Priyadarshini M, Vargas-Velazquez AM, 
            Frokjaer-Jensen C
  TITLE     Engineering rules that minimize germline silencing of transgenes in 
            simple extrachromosomal arrays in C. elegans.
  JOURNAL   Nat Commun. 2020 Dec 9;11(1):6300. doi: 10.1038/s41467-020-19898-0.
  PUBMED    33298957
REFERENCE   2  (bases 1 to 4464)
  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"; journalName: "Nat 
            Commun."; date: "2020-12-9"; volume: "11(1):6300. doi"; pages: " 
            10.1038/s41467-020-19898-0"
COMMENT     Sequence Label: pMDJ16 - pEXP[Peft-3 | mMaple3 (NLS) | tbb-2 UTR]
FEATURES             Location/Qualifiers
     source          1..4464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    37..57
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        58..657
                     /gene="eft-3"
                     /label=eft-3 promoter
                     /note="promoter for the C. elegans elongation factor 
                     1-alpha gene (Mitrovich and Anderson, 2000)"
     protein_bind    683..707
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             716..736
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1642..1713
                     /codon_start=1
                     /product="nuclear localization signal from the C. elegans 
                     EGL-13 transcription factor (Lyssenko et al., 2007)"
                     /label=EGL-13 NLS
                     /translation="SRRRKANPTKLSENAKKLAKEVEN"
     protein_bind    complement(1721..1745)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1750..2073
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(2077..2097)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(2105..2122)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2105..2121)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2114..2136)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2449..2471
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2509..2527)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2595..2699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2700..2768,2769..3560)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2918..2937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3731..4319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4220..4239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(4446..4464,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga aaagttggca
       61 cctttggtct tttattgtca acttccattg gttcttccat tgtttctgtt aaattaatga
      121 atttttcata aaataaagac attatacaat ataaaaatga agaatttatt gaaaataaac
      181 tgccagagag aaaaagtatg caacactccc gccgagagtg tttgaaatgg tgtacggtac
      241 attttcgtgc taggagttag atgtgcaggc agcaacgaga gggggagaga tttttttggg
      301 ccttgtgaaa ttaacgtgag ttttctggtc atctgactaa tcatgttggt tttttgttgg
      361 tttattttgt ttttatcttt gtttttatcc agattaggaa atttaaattt tatgaattta
      421 taatgaggtc aaacattcag tcccagcgtt tttcctgttc tcactgttta gtcgaatttt
      481 tattttaggc tttcaacaaa tgttctaact gtcttatttg tgacctcact ttttatattt
      541 ttttaatttt taaaaatatt agaagtttct aggataattt tttcgacttt tattctctct
      601 accgtccgca ctcttcttac ttttaaatta aattgttttt ttttcagttg ggaaacactt
      661 tgctcactcc gtagcagcca tggcaagttt gtacaaaaaa gcaggctcgg gatccccaaa
      721 gaagaagcgt aaggtcgtct ccaaggtaac acttagtttt tgttgctctg tttaaaaatt
      781 aattttaagg gagaggagac tatcatgtcc gtcatcaagc cagacatgaa gatcaagctc
      841 cgtatggagg gaaacgtcaa cggacacgcc ttcgtcatcg agggagaggg atccggaaag
      901 ccattcgagg gaatccaaac catcgacctc gaggtcaagg taagttttct tgagaccttt
      961 actaactaac taggtctcat agattttcag gagggagccc cactcccatt cgcctacgac
     1021 atcctcacca ccgccttcca ctacggaaac cgtgtcttca ccaagtaccc acgaaagatc
     1081 ccagactact tcaagcaatc cttcccaggt aagttatgta gagaccttaa tactcaacta
     1141 tggtctcaaa cattttcaga gggatactcc tgggagcgtt ccatgaccta cgaggacgga
     1201 ggaatctgca acgccaccaa cgacatcacc atggaggagg actccttcat caacaagatc
     1261 cacttcaagg gaaccaactt cccaccaaac ggaccagtca tgcaaaagcg taccgtcgga
     1321 tgggaggttt ccaccgagaa ggtaagttct ttagagacct aatcacatca atttggtctc
     1381 agtaattttc agatgtacgt ccgtgacgga gtcctcaagg gtgacgtcaa gatgaagctc
     1441 ctcctcaagg gaggatccca ctaccgttgc gacttccgta ccacctacaa ggtcaagcaa
     1501 aaggccgtca agctcccaaa ggcccacttc gtcgaccacc gtatcgagat cctctcccac
     1561 gacaaggact acaacaaggt caagctctac gagcacgccg tcgcccgtaa ctccaccgac
     1621 tccatggacg agctctacaa gtcccgtcgt cgtaaggcca acccaaccaa gctctccgag
     1681 aacgccaaga agctcgccaa ggaggtcgag aacggatcca acccagcttt cttgtacaaa
     1741 gtgggataaa tgcaaaatcc tttcaagcat tcccttcttc tctatcactc ttctttcttt
     1801 ttgtcaaaaa attctctcgc taatttattt gcttttttaa tgttattatt ttatgacttt
     1861 ttatagtcac tgaaaagttt gcatctgagt gaagtgaatg ctatcaaaat gtgattctgt
     1921 ctgatgtact ttcacaatct ctcttcaatt ccattttgaa gtgctttaaa cccgaaaggt
     1981 tgagaaaaat gcgagcgctc aaatatttgt attgtgttcg ttgagtgacc caacaaaaag
     2041 aggaaacttt attgtgccgc caagaaaaaa gtctcacaac tttattatac atagttgata
     2101 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     2161 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     2221 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
     2281 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt gcaatctgct
     2341 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     2401 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     2461 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     2521 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     2581 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2641 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2701 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2761 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2821 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2881 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2941 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3001 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3061 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3121 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3181 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3241 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3301 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3361 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3421 gcggtatcat tgcagcactg gggtcagatg gtaagccctc ccgtatcgta gttatctaca
     3481 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3541 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3601 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3661 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3721 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3781 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3841 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3901 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3961 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4021 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4081 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4141 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4201 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4261 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4321 acgccagcaa cgcggccttt ttacggttcc tggtcttttg ctggcctttt gctcacatgt
     4381 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4441 ataccagcgg ataacaattt caca
//
