LOCUS       sequence-486817-        4191 bp    DNA     circular SYN 22-SEP-2025
DEFINITION  Allows for rac-2(G12V)::mGFP entry into Gateway Vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4191)
  AUTHORS   Lamb R, Scales M, Watkins J, Werner M, Cherra SJ
  TITLE     A PDZ-RapGEF promotes synaptic development in C. elegans through a 
            Rap/Rac signaling pathway.
  JOURNAL   Development. 2025 Aug 1:dev.204678. doi: 10.1242/dev.204678.
  PUBMED    40748210
REFERENCE   2  (bases 1 to 4191)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Development. 2025 Aug 1:dev.204678. doi: 10.1242/dev.204678."
COMMENT     Sequence Label: pSJC318
FEATURES             Location/Qualifiers
     source          1..4191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(1321..1323,1324..1326,1327..2040)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLSDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1366..1387)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1627..1646)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1974..1995
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2070..2169)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2186..2205)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2187..2205)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2210..2226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2210..2226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2657..3448
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3541..4129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4030..4049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gaattcgccc ttggactagt catatgggta ccatgcaagc aatcaaatgt
      721 gtcgtcgttg gtgacgtagc cgtcggtaag acgtgtctcc tgctatcgta cactacaaac
      781 gcttttcccg gagaatatat tctaacggta ttcgacacct actcaacaaa tgtgatggtc
      841 gacggaaggc caataaatct cagcctatgg gacacagctg gacaggacga ttacgatcaa
      901 ttccgccacc tgtcatttcc acaaacagac gtattcctcg tatgctttgc actgaataat
      961 ccagcaagtt ttgagaatgt tcgtgcaaaa tggtatccag aagtatcaca tcattgccca
     1021 aatacaccga ttattttggt tggaactaaa gctgatttac gcgaggatcg agatactatt
     1081 gaacggctcc gtgaacgtcg gctccaacca gtgagccaca cccagggtta cgtgatggca
     1141 aaggaaatca aggcggtcaa gtacctggaa tgctcggcgc ttacccaaat tggattgaaa
     1201 caagttttcg atgaggcaat tcgtactggg ctcaccccgc cacaaacacc acaaacgaga
     1261 gccaaaaaga gcaattgcac ggtgggtacc agtggttccg gaggatctgg aggatctggt
     1321 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1381 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1441 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1501 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1561 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1621 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1681 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1741 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1801 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1861 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgtc cgacaaccac
     1921 tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1981 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtag
     2041 catatggcgg ccgcggaagg gcgaattcga cccagctttc ttgtacaaag ttggcattat
     2101 aaaaaataat tgctcatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     2161 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     2221 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     2281 tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt cgctgctacc
     2341 caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata
     2401 agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag
     2461 aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga
     2521 ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
     2581 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa
     2641 aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga
     2701 ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg
     2761 ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac
     2821 cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc
     2881 ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga
     2941 catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa
     3001 tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct
     3061 gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga
     3121 tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc
     3181 gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta
     3241 cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg
     3301 cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga
     3361 agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg
     3421 cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa
     3481 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3541 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3601 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3661 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3721 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3781 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3841 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3901 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3961 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4021 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4081 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4141 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
