LOCUS       sequence-489306-        7622 bp    DNA     circular SYN 15-OCT-2025
DEFINITION  Express rac-2(G12V) in all neurons..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7622)
  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 7622)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 15, 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: pREA3
FEATURES             Location/Qualifiers
     source          1..7622
                     /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    4722..4746
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5392..5416)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6258..6280)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6380..6399
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6477..6495)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6563..6667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6668..6736,6737..7528)
                     /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(6886..6905)
                     /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 tgttcttcta 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 gcgtgagcta 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 tattaacctt 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 aggtcaggca 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 ttcctggctc 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 ctgaacttgt 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 ctagaatttt 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 gctcctcttc 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 ctgcagaatc aagggcgaat tccagcacac tggcggccgt tactagagga
     4621 tccccgggat tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga
     4681 acattttcag gaggaccctt ggctagcgtc gacggtacat cacaagtttg tacaaaaaag
     4741 caggctccga attcgccctt ggactagtca tatgatgcaa gcaatcaaat gtgtcgtcgt
     4801 tggtgacgta gccgtcggta agacgtgtct cctgctatcg tacactacaa acgcttttcc
     4861 cggagaatat attctaacgg tattcgacac ctactcaaca aatgtgatgg tcgacggaag
     4921 gccaataaat ctcagcctat gggacacagc tggacaggac gattacgatc aattccgcca
     4981 cctgtcattt ccacaaacag acgtattcct cgtatgcttt gcactgaata atccagcaag
     5041 ttttgagaat gttcgtgcaa aatggtatcc agaagtatca catcattgcc caaatacacc
     5101 gattattttg gttggaacta aagctgattt acgcgaggat cgagatacta ttgaacggct
     5161 ccgtgaacgt cggctccaac cagtgagcca cacccagggt tacgtgatgg caaaggaaat
     5221 caaggcggtc aagtacctgg aatgctcggc gcttacccaa attggattga aacaagtttt
     5281 cgatgaggca attcgtactg ggctcacccc gccacaaaca ccacaaacga gagccaaaaa
     5341 gagcaattgc acggtgcttt aacatatggc ggccgcggaa gggcgaattc gacccagctt
     5401 tcttgtacaa agtggtgatg taccatggta ttgatatctg agctccgcat cggccgctgt
     5461 catcagatcg ccatctcgcg cccgtgcctc tgacttctaa gtccaattac tcttcaacat
     5521 ccctacatgc tctttctccc tgtgctccca ccccctattt ttgttattat caaaaaactt
     5581 ctcttaattt ctttgttttt tagcttcttt taagtcacct ctaacaatga aattgtgtag
     5641 attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa aattgtgctc cctcccccca
     5701 ttaataataa ttctatccca aaatctacac aatgttctgt gtacacttct tatgtttttt
     5761 acttctgata aatttttttg aaacatcata gaaaaaaccg cacacaaaat accttatcat
     5821 atgttacgtt tcagtttatg accgcaattt ttatttcttc gcacgtctgg gcctctcatg
     5881 acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt tttggaattt ttcaatcaag
     5941 tgaaagttta tgaaattaat tttcctgctt ttgctttttg gggtttcccc tattgtttgt
     6001 caagatttcg aggacggcgt ttttcttgct aaaatcacaa gtattgatga gcacgatgca
     6061 agaaagatcg gaagaaggtt tgggtttgag gctcagtgga aggtgagtag aagttgataa
     6121 tttgaaagtg gagtagtgtc tatggggttt ttgccttaaa tgacagaata cattcccaat
     6181 ataccaaaca taactgtttc tactagtcgg ccgtacgggc cctttcgtct cgcgcgtttc
     6241 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     6301 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     6361 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     6421 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcggcc ttaagggcct
     6481 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     6541 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6601 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6661 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6721 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6781 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6841 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6901 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6961 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7021 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7081 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7141 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7201 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7261 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7321 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7381 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7441 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7501 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7561 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7621 tc
//
