LOCUS       sequence-315633-        4458 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 4458)
  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 4458)
  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: pMDJ12 - pEXP[Peft-3 | mNeonGreen (NLS) | tbb-...
FEATURES             Location/Qualifiers
     source          1..4458
                     /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"
     regulatory      1081..1090
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1636..1707
                     /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(1715..1739)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1744..2067
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(2071..2091)
                     /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(2099..2116)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2099..2115)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2108..2130)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2443..2465
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2503..2521)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2589..2693
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2694..2762,2763..3554)
                     /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(2912..2931)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3725..4313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4214..4233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(4440..4458,1..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 aaggtgtcca aggtaacact tagtttttgt tgctctgttt aaaaattaat
      781 tttaagggag aggaggacaa catggcctcc ctcccagcca cccacgagct ccacatcttc
      841 ggctccatca acggagtcga cttcgacatg gtcggacaag gaaccggaaa cccaaacgac
      901 ggatacgagg agctcaacct caagtccacc aaggtaagtt ttcttgagac ctttactaac
      961 taactaggtc tcatagattt tcagggtgac ctccaattct ccccatggat actcgtccca
     1021 cacatcggat acggattcca ccaatacctc ccatacccag acggaatgtc cccattccaa
     1081 gccgccatgg tcgacgggtc cggataccaa gtccaccgta ccatgcaatt cgaggacgga
     1141 gcctccctca ccgtcaacta ccgttacacc tacgagggat cgcacatcaa ggtaagttat
     1201 gtagagacct ttactaacta actaggtctc aaacattttc agggagaggc ccaagtcaag
     1261 ggaaccggat tcccagccga cggaccagtc atgaccaact ccctcaccgc cgccgactgg
     1321 tgccgttcca agaagaccta cccaaacgac aagaccatca tctccacctt caagtggtcc
     1381 tacaccaccg gaaacggaaa gcgttaccgt tccaccgccc gtaccaccta caccttcgcc
     1441 aagccaatgg ccgccaacta cctcaaggta agttctttag agacctttac taactaacta
     1501 ggtctcagta attttcagaa ccaaccaatg tacgtcttcc gtaagaccga gctcaagcac
     1561 tccaagaccg agctcaactt caaggagtgg caaaaggcct tcaccgacgt catgggaatg
     1621 gacgagctct acaagtcccg tcgtcgtaag gccaacccaa ccaagctctc cgagaacgcc
     1681 aagaagctcg ccaaggaggt cgagaacgga tccaacccag ctttcttgta caaagtggga
     1741 taaatgcaaa atcctttcaa gcattccctt cttctctatc actcttcttt ctttttgtca
     1801 aaaaattctc tcgctaattt atttgctttt ttaatgttat tattttatga ctttttatag
     1861 tcactgaaaa gtttgcatct gagtgaagtg aatgctatca aaatgtgatt ctgtctgatg
     1921 tactttcaca atctctcttc aattccattt tgaagtgctt taaacccgaa aggttgagaa
     1981 aaatgcgagc gctcaaatat ttgtattgtg ttcgttgagt gacccaacaa aaagaggaaa
     2041 ctttattgtg ccgccaagaa aaaagtctca caactttatt atacatagtt gataattcac
     2101 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     2161 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     2221 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     2281 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtgcaatc tgctctgatg
     2341 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2401 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2461 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     2521 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     2581 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2641 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2701 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2761 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2821 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2881 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2941 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3001 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3061 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3121 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3181 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3241 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3301 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3361 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3421 tcattgcagc actggggtca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3481 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3541 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3601 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3661 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3721 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3781 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3841 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3901 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3961 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4021 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4081 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4141 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4201 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4261 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4321 gcaacgcggc ctttttacgg ttcctggtct tttgctggcc ttttgctcac atgttctttc
     4381 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgatacca
     4441 gcggataaca atttcaca
//
