LOCUS       sequence-217951-        7950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the C. elegans acy--1 P260S gain-of-function cDNA in 
            ventral cord cholinergic motor neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7950)
  AUTHORS   Schade MA, Reynolds NK, Dollins CM, Miller KG
  TITLE     Mutations that rescue the paralysis of Caenorhabditis elegans ric-8 
            (synembryn) mutants activate the G alpha(s) pathway and define a 
            third major branch of the synaptic signaling network.
  JOURNAL   Genetics. 2005 Feb;169(2):631-49. doi: 10.1534/genetics.104.032334. 
            Epub 2004 Oct 16.
  PUBMED    15489510
REFERENCE   2  (bases 1 to 7950)
  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"; doi: 
            "10.1534/genetics.104.032334"; journalName: "Genetics"; date: 
            "2005-02"; volume: "169"; issue: "2"; pages: "631-49"
COMMENT     Sequence Label: KG#84
FEATURES             Location/Qualifiers
     source          1..7950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    113..134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(149..154,155..172,173..179)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    187..203
                     /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     192..214
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     211..227
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     211..227
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5746..5768)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5868..5887
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5965..5983)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6051..6155
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6156..6224,6225..7016)
                     /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(6374..6393)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7187..7775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7676..7695
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7929..7946
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
       61 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      121 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      181 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
      241 agctgtaagt ttaaacatga tcttactaac taactattct catttaaatt ttcagagctt
      301 aaaaatggct gaaatcactc acaacgatgg atacgctaac aacttggaaa tgaaataagc
      361 ttgggctgca gaagcttttg gttttcacaa ttttctggtt tttgggattt ttgcgggaat
      421 tttgagagga gaaagttggg agagaagata ttggaatggt ccccggggag ctgtacattt
      481 atacaccaag gcaatcttcg taaaaaataa caatagagtc atccctcaga acagaaaaga
      541 aacccgcccg ggggctcaaa tacacatgtc atttttcttt tttcgcctga tttttagcct
      601 aaattagtgt ttagcaattt tttaaggtaa ctcggtggag tcatttgaac tttttaaact
      661 aaaatttcag aaaaaattct ttttttttgt caaaaaataa ttttcccgcc ggaatttttc
      721 aaaaaaataa tttttatttt atttttcaag ttccttgttt cctctttatt tttttatttt
      781 gcccccaaaa gttttacact aaaatattta attttcaggt aattcatcga aagtctttct
      841 attttccgca tctcttgttc aaggatcccc gggattggcc aaaggaccca aaggtatgtt
      901 tcgaatgata ctaacataac atagaacatt ttcaggagga cccttggcta gcgtcgacgg
      961 taccggtaaa aatggacgac gatgtcggtg aacgaacacc ggctctcggt ggttcatgtg
     1021 gaccatcagt tcgtgcacac agtagttccc caagacgtgt tccattattt gagagagcat
     1081 ctgctagatg gtggaatcct caatttcgat ctgcaacact agaagctcaa tattggaagt
     1141 gtagtttcag tcagttaaga gatcgatttc gttctggatt aatatatatt gccgttgtaa
     1201 ttgccgcttg gacactttat ttagctttat tcgatagaac ttttattcag cattggatcg
     1261 tttcactatg cctctgtgcc attattttcg cgatgtttgc cttcacggcg tgcgccgctc
     1321 aataccaaag gttctacatg ccaacctcat ttctatgcac ttttttgata tgccttgtca
     1381 cgttgctcat cttctccgca gagaatcaag ctgcgtttat gactccagtt gcaagtcttg
     1441 ccacaagttt tcaggtcgtt ctcctgatct acacagtcat tccactgcca ctctatttat
     1501 gtatactcat cggaataatc tactcaatcc tatttgagat tctcaacaag aacaaaattg
     1561 gtcttgaaga ggccggatac attaagcttg tccttcacgc tggtgtccat ttactcggtg
     1621 tccatttgtt cattttaact caagtgcgac agaggaaaac gtttctgaaa gtgggacaat
     1681 cgatgttggc aagaaaggat ctagagttgg agacacagtt taaagatcat atgattcagt
     1741 ctgtgatgtc taaaaaggta gctgatgaat tactgaaaga tgcatcagaa cttcgccgac
     1801 cttctgcttc caacgactca aactgccgaa cttcaaatgc tacacaagtt gaccagcccc
     1861 ttgccaagat ggtacctgaa taccgaaagt tccgtccatt tacgatgaat ctcatgacaa
     1921 atgtgtcgat tctgtttgcg gatattgctg ggtttactaa aatgtcatca aataaatctg
     1981 ccgacgaact tgtcaatctg ctcaatgatc tttttggaag attcgatact ttgtgtcgat
     2041 tgagaggatt agagaagatt tcaactttgg gagattgtta ctattgtgtt gctggatgtc
     2101 ctgaaccatg tgatgatcat gcatgcagaa cagtcgaaat gggtctcgac atgattgttg
     2161 caatccgcca atttgatatt gatcgtggac aagaagtgaa tatgcgtgtg ggaattcata
     2221 caggaaaagt aatgtgcgga atggttggaa caaaaagatt caaattcgat gtgttctcaa
     2281 acgatgtcac actggcaaat gagatggaat cttctggtgt tgctggaaga gttcacgttt
     2341 ccgaagcaac tgcaaaactt ttgaaagggc tttatgaaat tgaagaggga ccggattatg
     2401 atggtccatt gaggatgcaa gttcagggaa ctgaacgtcg tgttaaacca gaaagcatga
     2461 agactttctt tatcaaagga cgtattaatg atggtgtaga agaagaagtg atgcaagttc
     2521 aggaagttga gagccttcac agtcagaaaa gcagtaaaaa gtcgactcta aaacagaaat
     2581 gggcggaaaa gttgaaaatg aaccacacaa attcttatcc aatgcgagcc gctgcacgtg
     2641 aaggaggtgg aagtttgaga ataaaattgg cagagagaaa tagaagtaca cagttactgc
     2701 caaaagaatc aaattcaatt tgtataatgg aagataaccg aaagtcagca tcacttcaag
     2761 cattggcaac gaataatttt aatggaagta atacagatac aaataacact tatagtgaac
     2821 gtggagttgc tggaagtgtt agtaagaagt ctgtagctgg aagtgaaagt aatagtataa
     2881 aaggaagtag aagcagtgga ctacaattat ctctacaaga tggaaattct gatttaaatt
     2941 cagttggagg tttggatact gcaatcagtc accatcacaa tgcagcatct ctcactcgat
     3001 ttgatactga taacaatttt gatcagcgtt tggcaatggt aattggacaa ggagaaggag
     3061 gatttgataa aggattttgg aatcatcatg atagtttgaa taagtggact cttcgattta
     3121 atgaaaagga cgttgaagag gaatatcgag ctcattttgt ggattcatcg gaaagatata
     3181 cggcatcaaa gaaaggtcat gttgaaagac ataaagattt gatggaacaa ggtggagaaa
     3241 aagatggaat tactggatca acggtcaaca agtatagata ttcaggagtc ttcatcgata
     3301 taattgttgc cacgttgata tttgtaatat cgggtgcagt tgcaattatg tctgtccgtc
     3361 cattcccatt atcactcatt gcatacttcc catttgctgc tgcaattctc attcttacaa
     3421 ttgtccttat tggtcttcca cttctcgccc gtaaaaaatc attccaatgt gcaaatcaat
     3481 ggatgccacg tcatctcatc ggtcttcttc tgatctttct tccaatcggt gtcgcaatct
     3541 gtatcatgcc attgtgccaa agtggagact gtgcaaatgt aattctgaac taccgtcttg
     3601 cattcagtta tgtcaccatt cttgccatat tcgctcattg caacttctcc caattagctg
     3661 cctggccaaa aactactgct gcagtattta tcggacttct ccacattgct ggagttttct
     3721 actgcgaatt caatttgaaa cacttagttg aagagcaaga cacctgcaac gtgacagcaa
     3781 tcatgattcc accaattcgc aaaggtctga actacacaat tgcattgaac tcgacatctg
     3841 caagaacatt atctcaagat tttggatctc cattattcat ttgggagctg cttctggatg
     3901 tgatcctgtc gattgttcta gttgcattcc ttaattatca gttcgaaacc gcattccgta
     3961 tgtcattctt tggagatgtt caagctcgtc gggacactga aagaatgcaa atcgtgcgtg
     4021 atcaagccga ttggctcctg aataacgtga tcccagccca cgcagtagaa agtcttaaaa
     4081 cggataccaa atacagtgaa aaccatgaaa cagttggtgt actgttcgct tcaatcacta
     4141 attggaatga tatgtatgaa gagaatttcg aagggggcag agaattcttg agagttctca
     4201 atgaagttat tggtgacttt gatgagcttc tcgatcggcc ggacttcacg catattgaaa
     4261 aaatcaaaac aattgggcct gcgtatatgg cagcaagtgg attgaatcca gaacgaaaaa
     4321 agaatatgtt gcatccaaaa gagcatctat atcagatggt tgattttgca ttggcagttc
     4381 aacatgtact cagtgttttc aacgaggatt tgctcaattt tgattttgtt tgcaagttag
     4441 gacttaatat tggacccgtc actgctggtg tcattggaac aacaaagttg tactacgata
     4501 tctggggtga cactgtaaac attgctagcc gaatgtacag caccggagtt ttaaatcgca
     4561 ttcaagtatc acaacacaca cgagaatatc tcctggatcg gtatgaattc gagttccgcg
     4621 atcatattga agtcaaggga attgatggag gaatggatac ttatcttttg gttggtcgaa
     4681 aaggtgatgg aatacctcca tcaatcaagg ataaccaaga agacgaattt taaatctcga
     4741 ggagctcccg agatcttcag cgctcagcat tcgtagaatt gtaagtttaa actttctcat
     4801 actaactaac gcgctctatt taaattttca gaattccaac tgagcgccgg tcgctaccat
     4861 taccaacttg tctggtgtca aaaataatag gggccgctgt catcagagta agtttaaact
     4921 gagttctact aactaacgag taatatttaa attttcagca tctcgcgccc gtgcctctga
     4981 cttctaagtc caattactct tcaacatccc tacatgctct ttctccctgt gctcccaccc
     5041 cctatttttg ttattatcaa aaaacttctc ttaatttctt tgttttttag cttcttttaa
     5101 gtcacctcta acaatgaaat tgtgtagatt caaaaataga attaattcgt aataaaaagt
     5161 cgaaaaaaat tgtgctccct ccccccatta ataataattc tatcccaaaa tctacacaat
     5221 gttctgtgta cacttcttat gttttttact tctgataaat ttttttgaaa catcatagaa
     5281 aaaaccgcac acaaaatacc ttatcatatg ttacgtttca gtttatgacc gcaattttta
     5341 tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg ctcatcgtga aaaagttttg
     5401 gagtattttt ggaatttttc aatcaagtga aagtttatga aattaatttt cctgcttttg
     5461 ctttttgggg tttcccctat tgtttgtcaa gatttcgagg acggcgtttt tcttgctaaa
     5521 atcacaagta ttgatgagca cgatgcaaga aagatcggaa gaaggtttgg gtttgaggct
     5581 cagtggaagg tgagtagaag ttgataattt gaaagtggag tagtgtctat ggggtttttg
     5641 ccttaaatga cagaatacat tcccaatata ccaaacataa ctgtttccta ctagtcggcc
     5701 gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     5761 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     5821 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     5881 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5941 ataccgcatc aggcggcctt aagggcctcg tgatacgcct atttttatag gttaatgtca
     6001 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6061 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6121 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6181 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6241 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6301 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6361 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6421 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6481 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6541 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6601 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6661 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6721 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6781 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6841 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6901 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6961 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7021 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7081 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7141 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7201 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7261 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7321 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7381 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7441 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7501 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7561 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7621 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7681 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7741 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7801 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     7861 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     7921 ccgagcgcag cgagtcagtg agcgaggaag
//
