LOCUS       sequence-315624-        4646 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 4646)
  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 4646)
  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: pMDJ10 - pEXP[Peft-3 | tagBFP2 (NLS) | tbb-2 UTR]
FEATURES             Location/Qualifiers
     source          1..4646
                     /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"
     protein_bind    complement(1903..1927)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1932..2255
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(2259..2279)
                     /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(2287..2304)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2287..2303)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2296..2318)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2631..2653
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2691..2709)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2777..2881
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2882..2950,2951..3742)
                     /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(3100..3119)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3913..4501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4402..4421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(4628..4646,1..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 aaggtaagtt ttgaaatatt catattttta acttgaaaac taatatttaa
      781 attttttagg tcgccggcgg atcatcctcc acatcatcct ccacatcatc ctcatccgag
      841 ctcatcaagg taacacttag tttttgttgc tctgtttaaa aattaatttt aaggagaaca
      901 tgcacatgaa gctctacatg gagggaaccg tcgacaacca ccacttcaag tgcacctccg
      961 agggagaggg aaagccatac gagggaaccc aaaccatgcg tatcaaggtc gtcgagggag
     1021 gaccactccc attcgccttc gacatcctcg ccacctcctt cctctacgga tcgaagacct
     1081 tcatcaacca cacccaagga atcccagact tcttcaagca atccttccca ggtaagtttt
     1141 cttgagacct ttactaacta actaggtctc atagattttc agagggattc acctgggagc
     1201 gtgtcaccac ctacgaggac ggaggagtcc tcaccgccac ccaagacacc tccctccaag
     1261 acggatgcct catctacaac gtcaagatcc gtggagtcaa cttcacctcc aacggaccag
     1321 tcatgcaaaa ggtaagttat gtagagacct ttactaacta actaggtctc aaacattttc
     1381 agaagaccct cggatgggag gccttcaccg agacgctcta cccagccgac ggaggactgg
     1441 agggacgtaa cgacatggcc ctcaagctcg tcggaggatc gcacctcatc gccaacgcca
     1501 agaccaccta ccgttccaag aagccagcca agaacctcaa gatgccagtc tactacgtcg
     1561 actaccgtct ggagcgtaag gtaagttctt tagagacctt tactaactaa ctaggtctca
     1621 gtaattttca ggaggccaac aacgagacgt acgtcgagca acacgaggtc gccgtcgccc
     1681 gttactgcga cctcccatcc aagctcggac acaagctcaa ctcgggaact tctacgactt
     1741 cggtaacgtc atccggtact tccccagccg gctcccgtcg tcgtaaggta agttcataag
     1801 agacctttac taactaacta ggtctcagaa tttttcaggc caacccaacc aagctctccg
     1861 agaacgccaa gaagctcgcc aaggaggtcg agaacggatc caacccagct ttcttgtaca
     1921 aagtgggata aatgcaaaat cctttcaagc attcccttct tctctatcac tcttctttct
     1981 ttttgtcaaa aaattctctc gctaatttat ttgctttttt aatgttatta ttttatgact
     2041 ttttatagtc actgaaaagt ttgcatctga gtgaagtgaa tgctatcaaa atgtgattct
     2101 gtctgatgta ctttcacaat ctctcttcaa ttccattttg aagtgcttta aacccgaaag
     2161 gttgagaaaa atgcgagcgc tcaaatattt gtattgtgtt cgttgagtga cccaacaaaa
     2221 agaggaaact ttattgtgcc gccaagaaaa aagtctcaca actttattat acatagttga
     2281 taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     2341 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2401 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
     2461 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtgcaatctg
     2521 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     2581 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2641 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     2701 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     2761 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2821 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2881 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2941 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     3001 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3061 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3121 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3181 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3241 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3301 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3361 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3421 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3481 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3541 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3601 tcgcggtatc attgcagcac tggggtcaga tggtaagccc tcccgtatcg tagttatcta
     3661 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3721 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3781 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3841 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3901 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3961 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4021 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4081 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4141 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4201 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4261 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4321 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4381 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4441 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4501 aaacgccagc aacgcggcct ttttacggtt cctggtcttt tgctggcctt ttgctcacat
     4561 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4621 tgataccagc ggataacaat ttcaca
//
