LOCUS       sequence-486824-        4173 bp    DNA     circular SYN 22-SEP-2025
DEFINITION  Allows for rap-1(G12V)::mGFP entry into Gateway Vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4173)
  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 4173)
  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: pSJC316
FEATURES             Location/Qualifiers
     source          1..4173
                     /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(1303..1305,1306..1308,1309..2022)
                     /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(1348..1369)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1609..1628)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1956..1977
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2052..2151)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2168..2187)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2169..2187)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2192..2208)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2192..2208)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2639..3430
                     /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      3523..4111
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4012..4031
                     /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 ccatgcggga gtataagatt
      721 gttgtgctcg gatctgtcgg agtaggaaaa tccgcactga cggtacaatt cgtgcaggga
      781 attttcgtag agaagtatga cccaacaatt gaggatagtt acagaaagca agtggaagtc
      841 gacggccagc aatgtatgct tgagatactg gatacagcag gaacggagca attcacagcg
      901 atgcgtgatt tgtacatgaa gaatggacaa ggatttgttc tcgtttattc aattactgct
      961 caatcgacat tcaatgatct tatggatctt cgtgatcaga ttcttcgtgt caaagatacg
     1021 gacgaggttc caatgatttt ggtaggaaat aagtgtgatt tggaagacga acgagttgtt
     1081 ggaaaagatc agggacagaa tctcgccagg cagtttggat cagcattcct tgaaacctct
     1141 gcaaaagcaa agattaacgt cagcgaggtg ttctacgacc tcgttcgtca aataaaccga
     1201 cgttatcccg aatctggtcg acgccaggga caatcaaata agcagtgctg ctcgtgtgtc
     1261 atcatgggta ccagtggttc cggaggatct ggaggatctg gtatggtgag caagggcgag
     1321 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1381 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1441 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     1501 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     1561 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1621 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1681 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1741 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     1801 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1861 acccccatcg gcgacggccc cgtgctgctg tccgacaacc actacctgag cacccagtcc
     1921 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1981 gccgccggga tcactctcgg catggacgag ctgtacaagt agcatatggc ggccgcggaa
     2041 gggcgaattc gacccagctt tcttgtacaa agttggcatt ataaaaaata attgctcatc
     2101 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2161 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2221 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct
     2281 ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg
     2341 cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa
     2401 gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
     2461 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca
     2521 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
     2581 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat
     2641 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga
     2701 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg
     2761 cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac
     2821 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga
     2881 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta
     2941 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat
     3001 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca
     3061 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga
     3121 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg
     3181 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa
     3241 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca
     3301 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc
     3361 gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt
     3421 cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3481 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3541 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3601 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3661 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3721 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3781 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3841 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3901 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3961 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4021 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4081 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4141 cctggccttt tgctggcctt ttgctcacat gtt
//
