LOCUS       sequence-483367-        7680 bp    DNA     circular SYN 19-AUG-2025
DEFINITION  Express rap-1(G12V) in all neurons..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7680)
  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 7680)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 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: pREA6
FEATURES             Location/Qualifiers
     source          1..7680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      77..665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     566..585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     819..836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    953..974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(989..994,995..1012,1013..1019)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1027..1043
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1032..1054
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1051..1067
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1051..1067
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4758..4782
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5442..5466)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6316..6338)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6438..6457
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6535..6553)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6621..6725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6726..6794,6795..7586)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6944..6963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
       61 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
      121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
      241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg
      481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      781 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      841 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      901 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      961 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     1021 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     1081 agcttgcatg cctgcaggtc gactctagcc cccacacaca tcgctgccgg aatagatgat
     1141 cccatcgtag ttcgataatc tcgtgacact cgtttccgat acccccttat atcagcacac
     1201 attcagtcat cataccacaa aatgttgtga tagtgatagg caaaactacc gtatttcctc
     1261 tattaatctt gcatgcaaga ctaattttcg attaacccgt aggggtgcaa gactaataga
     1321 gactgcaaga ctattagagg ctgaaatact aattttcgta tgctcaataa ttttggaaat
     1381 tggcctattt tttgtagaaa cttgataccg tttaaacaag gaaaaataca cactttttaa
     1441 tatttcatca ataatttgaa cgattttgtg attttaagtt caatttgccc aaaaaaagac
     1501 aatttttctg acgttaccgc aaagtaatgc tggtcaggca aaaagagcgg tgcaaaaata
     1561 taagagactg caatactaat agaggaaata cggtaattga attttagtga aacttgcgac
     1621 agttttcctc atttttttgt tattccggca tacgagtgtg gcatacgagt gaggtcatct
     1681 tttgtttctt ccgtttcttc atcgcttttg aaaaaaaatg ttgaagaaaa cttcgtagcc
     1741 gatagccgga taatgtttgg cacggcgttc caacatatat cattggaaat tttttaagat
     1801 ttttgcgaaa aatcacattc ttcacgatga gaacacgtta ttgaaggaat ataaatcaag
     1861 aataacatat agttatattt ctctattact ttaacgttaa atatgagcaa atttgagcat
     1921 tttgattgcg atgaaaagca gaatcgagtc aactgaaatc cgttcaaaaa cctaagctcg
     1981 gttcgcccga aagtcgattt ttccaatagc cgaacagcac tgcttcttta ttttccaggt
     2041 atttttgggg tgtaagaatt atatcttctt tgtttgggta tttacgtttc cgaattccct
     2101 ttcttgatct gtcgctattt ttcagaaacc agaacagttt atcctttttc atgagaacct
     2161 gaatagctca aaaacccgtt taatttcttt tcatcatctc atttgaaact ttcctagtct
     2221 tcttaatgat ttccaggctc ctcctacatt tgtatctcaa gatgcataca ctttatttcc
     2281 gctccaattc gaatcatcta cttctctttt ttcttcaact ttctactttt ttccattctt
     2341 ttctttcaaa tgttcgacgt cttcaagtga ctgctcttca ttctcctcat ttttctcgaa
     2401 tttcattctt gtctcttttt tgctcaaaaa tggaaaatga aagtgacgtg agatttggac
     2461 ggcgggacac gggggcagta gaagcagcaa aaaggagaga aagaggacac aaataagaag
     2521 aacgaattca aaaataagcg gagaggagct atttccgtca attctacctc cccaatcttc
     2581 atcaattcgg ctcaattgaa tgacgtcaca gggagataaa ccgtttggat gagcgccgtc
     2641 catcactgac gccatcccgt ttgggacaag aaaagagaaa aaagagcaca aagtttttgg
     2701 tgacggatct tgtcaatcat atgaaagttg ttctgattga ttgtcagttt ttttcctact
     2761 tttttggatt ctatccactt ctgaactttt gacaagtttc aaactttctg aatcattttc
     2821 tatgcatttt cctggaattc tttttatgta aaatatgaaa tagaatgttt ttgaattcaa
     2881 gttctgcttt tttcttcttt tttgttctgt tcgggcttgg gtatgctttt tttaaaaatt
     2941 attttgcaca tcgaccaata agtgcgcaac ttataaaatt aatttatttt tgttaatttt
     3001 tgaaatactt gtattgcttt aagtgatctg acctcgccct gagctttcca cgtagttatc
     3061 aaatacaaat cctccaaggg taacgtacct atattactga tctttataat aactttatca
     3121 cctgtccagt tccagaggaa ttctgttaag cttataggca cagaaggagt catttctgct
     3181 ggtttttaat gatccaaatc tttatttcaa gtaaaaaact gaacacttgc gaataaaact
     3241 atcagattaa ccattcacca aaaatgtgtt tgaatctaaa actttctcag tattccaaat
     3301 atagaaataa ataaccacga cattgctaaa atctgtctga attgtgtact ccttaccgtg
     3361 aagtaataat ggatataatg aatcgtttga aatgaatgat cagcacattt ttggtgaaag
     3421 atcacaaata aggaataagc gacggaaaat aaaacgatta tttcggatca aaaatttgtt
     3481 gaagatcata tacactcgag accaagatta ttctagacaa ttttcaaatt ggcttctttg
     3541 tttgcaaatc ttcataataa tctgtgaagt ttggaaattt gaatttttaa tcttttttca
     3601 tagattatag tttttatttc tttgcaaaac tatattaaaa accgatgcat tgttttaggg
     3661 aaattaatga gccttttgtt caacactaaa aacaataaaa ttaaaatttt ggcttcatca
     3721 tttgaccttt tttaagttcg aaaacttttc tcgtatttct gaaccgccaa ttttttcaca
     3781 catctctaga cttttggtgc ccgttccaga aagttaagta attgctattc taagaaagtt
     3841 ctcaacattg ttttttagtt ctgattgaat tctgatgttc caggaatata ttttaaattt
     3901 aatattcttt gcactatttc tatactaact aaataataaa tagtctaagt atgttcaatg
     3961 agaacaaaaa agctctatct atatttttct atcctatttc atttatatcc ttttcatttt
     4021 gaactcacca gcttctcttc ttcttcttgt ccatatactt cttaaaagtc tcgaacttct
     4081 ctctctcact ctccatcaat ttctttatgg ctaccgcttg cgctctgccg cttccgaaga
     4141 aagaaggcgt tggcagagct ctcaattcta ttttttctcg ccgtaggcta tcgatttctc
     4201 agcttctctt ctctcttcct cactccacca ccaccacttc gggcttcttc ttttcttcat
     4261 tttcgtcttc ttcttcatcc tcttcttttt ttttcagttc cccctcactc ctccccttat
     4321 atgcgcgtgc gagaggggtg caaaagcagc gcgcatccga gaattgaaac gagaaaacgg
     4381 agacgcagca gcagttcgtc ctcagaaaaa tagccagtaa aaagagaaaa agatagagag
     4441 agacctatcg cattttattt tcaaattgca attcctgcaa ttcatgtgtg cgtgtgccca
     4501 ttttcaattc ttcccgttat ttttcagacc attaccaacg ttctttctag atcttgagac
     4561 aatcttcctt ctgctcaatc gttcgtcgtg aagacggcat cgacgactgc agcgataagg
     4621 gcgaattcca gcacactggc ggccgttact agaggatccc cgggattggc caaaggaccc
     4681 aaaggtatgt ttcgaatgat actaacataa catagaacat tttcaggagg acccttggct
     4741 agcgtcgacg gtacatcaca agtttgtaca aaaaagcagg ctccgaattc gcccttggac
     4801 tagtcatatg caccataaca gatcttcaaa aaaagaatgc gggagtataa gattgttgtg
     4861 ctcggatctg tcggagtagg aaaatccgca ctgacggtac aattcgtgca gggaattttc
     4921 gtagagaagt atgacccaac aattgaggat agttacagaa agcaagtgga agtcgacggc
     4981 cagcaatgta tgcttgagat actggataca gcaggaacgg agcaattcac agcgatgcgt
     5041 gatttgtaca tgaagaatgg acaaggattt gttctcgttt attcaattac tgctcaatcg
     5101 acattcaatg atcttatgga tcttcgtgat cagattcttc gtgtcaaaga tacggacgag
     5161 gttccaatga ttttggtagg aaataagtgt gatttggaag acgaacgagt tgttggaaaa
     5221 gatcagggac agaatctcgc caggcagttt ggatcagcat tccttgaaac ctctgcaaaa
     5281 gcaaagatta acgtcagcga ggtgttctac gacctcgttc gtcaaataaa ccgacgttat
     5341 cccgaatctg gtcgacgcca gggacaatca aataagcagt gctgctcgtg tgtcatcatg
     5401 tgattatctt tacatatggc ggccgcggaa gggcgaattc gacccagctt tcttgtacaa
     5461 agtggtgatg taccatggta ttgatatctg agctccgcat cggccgctgt catcagatcg
     5521 ccatctcgcg cccgtgcctc tgacttctaa gtccaattac tcttcaacat ccctacatgc
     5581 tctttctccc tgtgctccca ccccctattt ttgttattat caaaaaaact tcttcttaat
     5641 ttctttgttt tttagcttct tttaagtcac ctctaacaat gaaattgtgt agattcaaaa
     5701 atagaattaa ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc cattaataat
     5761 aattctatcc caaaatctac acaatgttct gtgtacactt cttatgtttt ttttacttct
     5821 gataaatttt ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt atcatatgtt
     5881 acgtttcagt ttatgaccgc aatttttatt tcttcgcacg tctgggcctc tcatgacgtc
     5941 aaatcatgct catcgtgaaa aagttttgga gtatttttgg aatttttcaa tcaagtgaaa
     6001 gtttatgaaa ttaattttcc tgcttttgct ttttgggggt ttcccctatt gtttgtcaag
     6061 agtttcgagg acggcgtttt tcttgctaaa atcacaagta ttgatgagca cgatgcaaga
     6121 aagatcggaa gaaggtttgg gtttgaggct cagtggaagg tgagtagaag ttgataattt
     6181 gaaagtggag tagtgtctat ggggtttttg ccttaaatga cagaatacat tcccaatata
     6241 ccaaacataa ctgtttccta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg
     6301 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     6361 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     6421 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     6481 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg
     6541 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     6601 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6661 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6721 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6781 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6841 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6901 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6961 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7021 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7081 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7141 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7201 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7261 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7321 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7381 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7441 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7501 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7561 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7621 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
//
