LOCUS       sequence-375296-        8604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Punc-17::GtACR2::wCherry unc-54 3' UTR C.elegans neuron expression 
            of GtACR2 RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8604)
  AUTHORS   Lu Y, Ahamed T, Mulcahy B, Meng J, Witvliet D, Guan SA, Holmyard D, 
            Hung W, Wen Q, Chisholm AD, Samuel ADT, Zhen M
  TITLE     Extrasynaptic signaling enables an asymmetric juvenile motor circuit
            to produce symmetric undulation.
  JOURNAL   Curr Biol. 2022 Nov 7;32(21):4631-4644.e5. doi: 
            10.1016/j.cub.2022.09.002. Epub 2022 Sep 30.
  PUBMED    36182701
REFERENCE   2  (bases 1 to 8604)
  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: 
            "10.1016/j.cub.2022.09.002"; journalName: "Curr Biol"; date: 
            "2022-11-7- 7"; volume: "32"; issue: "21"; pages: "4631-4644.e5"
COMMENT     Sequence Label: pJH4330
FEATURES             Location/Qualifiers
     source          1..8604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      118..127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6136..6158)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6258..6277
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6355..6373)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6441..6545
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6546..6614,6615..7406)
                     /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(6764..6783)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7577..8165
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8066..8085
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8319..8336
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8453..8474
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8489..8494,8495..8512,8513..8519)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8527..8543
                     /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     8532..8554
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8551..8567
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8551..8567
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 actaactaac tattctcatt taaattttca gagcttaaaa atggctgaaa tcactcacaa
       61 cgatggatac gctaacaact tggaaatgaa ataagcttgg gctgcagcat gctcccggcc
      121 gccatggccg cgggattaca ccaatcattt ctcccctccg aaatgtgtag tcgtcatgac
      181 aaagtggtga cactggatta cggtaggctc cgcccacttc ttggaaaagt gacgtgggct
      241 cggtgagccc taaagctttt ggttttcaca attttctggt ttttgggatt tttgcgggaa
      301 ttttgagagg agaaagttgg gagagaagat attggaatgg tccccgggga gctgtacatt
      361 tatacatcta gactgaagct ggggtattgc aaaaagagaa aagcctatca ctttctttcc
      421 atcctgcata cttgccaaaa agttttggct gagcttattt ttaaaattgg aattttttgg
      481 aattttcaga aaagaaaaag ttttttcttt ttttttaatt taaaaaattt ttgctattta
      541 ttgcatggta ctacagaaaa acttgaatga aataaaaaat ttggaaaatt caagaaaact
      601 ttttttttca tagtttattt attcgagttt taatgtggct tttattttgt tcggtttttg
      661 ttttaaaaag attttataca aaatttcaag aatttacaat ttttccgttg taaaaaatcc
      721 aaaaaaaaat ccaaaaaaat agttcagacc tttttttaaa ttttaaattt aaaaattcaa
      781 taccaaaaaa ttaaaaaaaa ttagtttttc cgagaataaa ggaaaaatgt tgaaaaaata
      841 ttatatataa aatttccaga aaatgtgaaa actccaaaaa actgcagact tttccccaaa
      901 ctagctattt tttgaattca ttacctttaa aggtggagta ccgaaattta agactttgct
      961 tttttagacc caaaatggtc caaaactacc aaatttcgta atgagacgtt ctgaaaattt
     1021 ttcaaaaaaa tgttatggga gctcaaagtt tttgaaaaaa tggtatattt ttagctaatt
     1081 ttaaccactt cttggcgtcg cagcggttgg aactcaattt taattaaatt atcatccttc
     1141 aatgtccttc aatctttgca atttcaatcg accatacgcg ttcagagcat tttttttttt
     1201 cgtttttcct cccgtccagc acatatttaa atattctctc tccccctccc ctggttccta
     1261 aaaacttggg aaaggggaat ttcgaaggag gtggaagctt cttcttctcc ccgagcacac
     1321 acacacatcc tctcttcaaa atctttaaga aatagtgcgt gtgaatgact agacgctgaa
     1381 gctaacttgg tttccgcgaa caatcccaat cagtgcctgt gccgaggctt cggagaagaa
     1441 gagattgacg tcgtcttttt ctttttttgt gaaacagaat aaaaaagaat acgaaatgga
     1501 ctttttcaca cattttcgtt acttttccag aaaaaaacga agctctagcc atggaagttt
     1561 gaattgaaaa cctaaacccc gatgtgtatt catttgagct taagcctaag cctaattcta
     1621 cgcctaagca taagcctaag cctatgttca agcctaagcc aaattgtaga acgttaccaa
     1681 aagatcaaac ctatttttcc tacagaaaag gtctaatgat aactgttaag cgagagacaa
     1741 ggagagatat atatcaaaaa ctagcccaaa ctgtgattaa catctattaa ctgacctcaa
     1801 aaaaaaaaat agtagagatt aaatactgct caaaaagttt cttggaaaag taaattgtta
     1861 ggtttctggg gcctgtaaaa ctggaccgag ttttgaagaa tttttttaac ttttcaaagt
     1921 ttggcgtcac tgaaagcttt atattgtaca tgttccacca attttctttt gtatccgttt
     1981 tttctgtctc aaaacggagc tcgagcccat gacgtgtgga taccgtgaga atccgcccaa
     2041 ttccttcaca tcgatctcat atttctttcc cagtttatat tgaattcccg ttgaattgcg
     2101 gctttgcagg ggagaagatg tgcgtcttta tatttttttt tctatcctta cccgccgtga
     2161 atccggctcg aattgagcca atttttatga atagtagagc taattgactt cgtctcccta
     2221 gaattgatga ggtaaagctc gtggggagat atggagattt ttcgattttt tttccagtaa
     2281 ttttcagttc aaaatttgag cttttgagca ggagttttga gatttgtgcc aaaatttcct
     2341 attatttttt caaatatttt tgcttttgta aaaaaggtgt agtagaagtt ttgtagtgcc
     2401 actatttttt ttttgaaaat tatgttttgg cggaaagttc tttaaaaaag ttaatcaaaa
     2461 tttttggcgc tttagtcaaa ttaattttct ggcaaaaata ttagtaattt tggcgtgagt
     2521 tcaattttcc cattcttaaa atttcacaca attaagaatt ttaagatttg ggtttttttt
     2581 tcaacaaaaa atcaaatatt tacatgaaaa ttcggcggga aataaaaaaa aatattccga
     2641 ttcaatgtgt gaataacttt ttgcggggaa ttcaaatttg cattatttaa acttttttta
     2701 agctcgaaaa attgaaattc ttaaataaat aaataaaatt ttcctcgaat tttgaatgtt
     2761 tcgatttttt agcgttattt tttcaaaatt tccactaaaa ctctagttct aaagttctaa
     2821 aactttaaaa acttcctaat gttgtggtct tgtgtttttt tggcagcgat ttccaaattt
     2881 ctgaaattcc aacaaaaaaa ccactacgtc agtttgcata tccaaccccc ccccccctct
     2941 gagccaatca atcatgtgca ttttgaattg gcgccaaggc aatcttcgta aaaaataaca
     3001 atagagtcat ccctcagaac agaaaagaaa cccgcccggg ggctcaaata cacatgtcat
     3061 ttttcttttt tcgcctgatt tttagcctaa attagtgttt agcaattttt taaggtaact
     3121 cggtggagtc atttgaactt tttaaactaa aatttcagaa aaaattcttt ttttttgtca
     3181 aaaaataatt ttcccgccgg aatttttcaa aaaaataatt tttattttat ttttcaagtt
     3241 ccttgtttcc tctttatttt tttattttgc ccccaaaagt tttacactaa aatatttaat
     3301 tttcaggtaa ttcatcgaaa gtctttctat tttccgcatc tcttgttcaa gggattggcc
     3361 aaaggaccca aaggtatgtt tcgaatgata ctaacataac atagaacatt ttcaggagga
     3421 cccttggcta gcatggcctc ccaggtcgtg tacggcgagt gggccagcac ccacacagag
     3481 tgctacaaca tgagcaggat cgacagcacc ttcgtgagcc tgctgcagct ggtgtgggcc
     3541 gtggtgagcg gatgccagac catcttcatg atcagcaggg cccccaaggt gccctgggag
     3601 agcgtgtacc tgcccttcgt ggagtccatc acatacgccc tggccagcac cggcaatggc
     3661 acactgcaga tgagggacgg caggttcttt ccctggtcca ggatggccag ctggctgtgc
     3721 acatgcccca ttatgctggg ccagatctcc aacatggctc tggtgaagta caagagcatc
     3781 cctctgaacc ccatcgccca ggccgccagc attatcaggg tggtgatggg cattaccgct
     3841 accatctccc ccgccgagta catgaagtgg ctgttcttct tcttcggcgc tacctgcctg
     3901 gtcttcgaat acagcgtggt gttcaccatc ttccaggtgg gactgtacgg cttcgagtcc
     3961 gtgggcaccc ctctggccca gaaggtggtg gtgaggatca agatgctgag gctgatcttc
     4021 ttcatcgcct ggaccatgtt ccccatcgtg tggctgattt cccccaccgg cgtgtgcgtg
     4081 atccacgaga acgtgagcgc catcctgtat ctgctcgccg acggcctgtg caagaacacc
     4141 tacggcgtga tcctgtggtc caccgcctgg ggagtgctcg agggaaagtg ggacccggct
     4201 tgcctccctg gccaggagaa gcctgaggcc gacgacccct tcggactgaa ccacgagaag
     4261 aatgcccccc ccaacgacga ggtcaacatc agaatgttcg gcagggcggc cgccgcggta
     4321 ccgggaggag gaaccatggt ctcaaagggt gaagaagata acatggcaat tattaaagag
     4381 tttatgcgtt tcaaggtgca tatggaggga tctgtcaatg ggcatgagtt tgaaattgaa
     4441 ggtgaaggag aaggccgacc atatgaggga acacaaaccg caaaactaaa ggtaagttta
     4501 aacatatata tactaactaa ccctgattat ttaaattttc aggtaactaa aggcggacca
     4561 ttaccattcg cctgggacat cctctctcca cagttcatgt atggaagtaa agcttatgtt
     4621 aaacatccgg cagatatacc agattatttg aaactttcat tcccggaggg ttttaagtgg
     4681 gaacgcgtaa tgaattttga agacggagga gttgttacag tgacgcaaga ctcaaggtaa
     4741 gtttaaacag ttcggtacta actaaccata catatttaaa ttttcagcct ccaagatgga
     4801 gaatttattt ataaagtcaa acttcgagga acgaatttcc cctcggatgg atctgttatg
     4861 cagaagaaga ctatgggatg ggaagcttca agtgaaagaa tgtaccctga agacggtgct
     4921 cttaagggag agattaaaca acgtcttaaa ttgaaagatg gaggacatta cgatgctgag
     4981 gtaagtttaa acatgatttt actaactaac taatctgatt taaattttca ggtgaagaca
     5041 acttacaaag ccaaaaaacc agttcagctg ccaggagcgt acaatgttaa tattaaactg
     5101 gatatcacct cccacaacga ggattacact atcgttgagc aatatgaaag agctgaaggg
     5161 cggcactcga caggtggcat ggatgaattg tataagttct gctacgagaa cgaggtctaa
     5221 gaattccaac tgagcgccgg tcgctaccat taccaacttg tctggtgtca aaaataatag
     5281 gggccgctgt catcagagta agtttaaact gagttctact aactaacgag taatatttaa
     5341 attttcagca tctcgcgccc gtgcctctga cttctaagtc caattactct tcaacatccc
     5401 tacatgctct ttctccctgt gctcccaccc cctatttttg ttattatcaa aaaacttctc
     5461 ttaatttctt tgttttttag cttcttttaa gtcacctcta acaatgaaat tgtgtagatt
     5521 caaaaataga attaattcgt aataaaaagt cgaaaaaaat tgtgctccct ccccccatta
     5581 ataataattc tatcccaaaa tctacacaat gttctgtgta cacttcttat gttttttact
     5641 tctgataaat ttttttgaaa catcatagaa aaaaccgcac acaaaatacc ttatcatatg
     5701 ttacgtttca gtttatgacc gcaattttta tttcttcgca cgtctgggcc tctcatgacg
     5761 tcaaatcatg ctcatcgtga aaaagttttg gagtattttt ggaatttttc aatcaagtga
     5821 aagtttatga aattaatttt cctgcttttg ctttttgggg tttcccctat tgtttgtcaa
     5881 gatttcgagg acggcgtttt tcttgctaaa atcacaagta ttgatgagca cgatgcaaga
     5941 aagatcggaa gaaggtttgg gtttgaggct cagtggaagg tgagtagaag ttgataattt
     6001 gaaagtggag tagtgtctat ggggtttttg ccttaaatga cagaatacat tcccaatata
     6061 ccaaacataa ctgtttccta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg
     6121 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     6181 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     6241 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     6301 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg
     6361 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     6421 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6481 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6541 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6601 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6661 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6721 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6781 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6841 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6901 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6961 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7021 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7081 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7141 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7201 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7261 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7321 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7381 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7441 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7501 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7561 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7621 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7681 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7741 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7801 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7861 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7921 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7981 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8041 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8101 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8161 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     8221 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     8281 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     8341 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     8401 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     8461 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     8521 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     8581 agctgtaagt ttaaacatga tctt
//
