LOCUS       sequence-315747-        4482 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 4482)
  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 4482)
  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: pMDJ15 - pEXP[Peft-3 | mCardinal (NLS) | tbb-2...
FEATURES             Location/Qualifiers
     source          1..4482
                     /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             1660..1731
                     /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(1739..1763)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1768..2091
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(2095..2115)
                     /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(2123..2140)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2123..2139)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2132..2154)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2467..2489
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2527..2545)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2613..2717
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2718..2786,2787..3578)
                     /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(2936..2955)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3749..4337
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4238..4257
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(4464..4482,1..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 aggagctcat caaggagaac atgcacatga agctctacat ggagggaacc
      841 gtcaacaacc accacttcaa gtgcaccacc gagggagagg gaaagccata cgagggaacc
      901 caaacccaac gtatcaaggt cgtcgaggga ggaccactcc cattcgcctt cgacatcctc
      961 gccacctgct tcatgtacgg atcgaaggta agttttcttg agacctttac taactaacta
     1021 ggtctcatag attttcagac cttcatcaac cacacccaag gaatcccaga cttcttcaag
     1081 caatccttcc cagagggatt cacctgggag cgtgtcacca cctacgagga cggaggagtc
     1141 ctcaccgtca cccaagacac ctccctccaa ggtaagttat gtagagacct ttactaacta
     1201 actaggtctc aaacattttc agacggatgc ctcatctaca acgtcaagct ccgtggagtc
     1261 aacttcccat ccaacggacc agtcatgcaa aagaagaccc tcggatggga ggccaccacc
     1321 gagacgctct acccagccga cggaggactc gagggacgtt gcgacatggc cctcaagctc
     1381 gtcggaggag gacacctcca ctgcaacctc aaggtaagtt ctttagagac ctttactaac
     1441 taactaggtc tcagtaattt tcagaccacc taccgttcca agaagccagc caagaacctc
     1501 aagatgccag gagtctactt cgtcgaccgt cgtctcgagc gtatcaagga ggccgacaac
     1561 gagacgtacg tcgagcaaca cgaggtcgcc gtcgcccgtt actgcgacct cccatccaag
     1621 ctcggacaca agctcaacgg aatggacgag ctctacaagt cccgtcgtcg taaggccaac
     1681 ccaaccaagc tctccgagaa cgccaagaag ctcgccaagg aggtcgagaa cggatccaac
     1741 ccagctttct tgtacaaagt gggataaatg caaaatcctt tcaagcattc ccttcttctc
     1801 tatcactctt ctttcttttt gtcaaaaaat tctctcgcta atttatttgc ttttttaatg
     1861 ttattatttt atgacttttt atagtcactg aaaagtttgc atctgagtga agtgaatgct
     1921 atcaaaatgt gattctgtct gatgtacttt cacaatctct cttcaattcc attttgaagt
     1981 gctttaaacc cgaaaggttg agaaaaatgc gagcgctcaa atatttgtat tgtgttcgtt
     2041 gagtgaccca acaaaaagag gaaactttat tgtgccgcca agaaaaaagt ctcacaactt
     2101 tattatacat agttgataat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     2161 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     2221 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     2281 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     2341 ctctcagtgc aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     2401 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     2461 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     2521 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2581 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2641 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2701 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2761 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2821 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2881 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2941 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3001 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3061 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3121 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3181 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3241 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3301 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3361 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3421 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gtcagatggt aagccctccc
     3481 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3541 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3601 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3661 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3721 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3781 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3841 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3901 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3961 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4021 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4081 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4141 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4201 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4261 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4321 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gtcttttgct
     4381 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4441 ccgcctttga gtgagctgat accagcggat aacaatttca ca
//
