LOCUS       sequence-216868-        8396 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Emerald-RAB-3 in the cholinergic motor neuron DA9 for 
            imaging synaptic vesicles in a single neuron in a living animal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8396)
  AUTHORS   Edwards SL, Morrison LM, Manning L, Stec N, Richmond JE, Miller KG
  TITLE     Sentryn Acts with a Subset of Active Zone Proteins To Optimize the 
            Localization of Synaptic Vesicles in Caenorhabditis elegans.
  JOURNAL   Genetics. 2018 Nov;210(3):947-968. doi: 10.1534/genetics.118.301466.
  PUBMED    30401765
REFERENCE   2  (bases 1 to 8396)
  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.118"; journalName: "Genetics"; date: "2018-11"; 
            volume: "210"; issue: "3"; pages: "947-968"
COMMENT     Sequence Label: KG#901
FEATURES             Location/Qualifiers
     source          1..8396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2..18
                     /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     7..29
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     26..42
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     26..42
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(3587..3589,3590..3592,3593..4303)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3632..3653)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3893..3912)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4240..4261
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6003..6025)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6125..6144
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6226..6244)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6312..6416
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6417..6485,6486..7277)
                     /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(6635..6654)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7448..8036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7937..7956
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8190..8207
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8324..8345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8360..8365,8366..8383,8384..8390)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt
       61 gcatgcctga cactaagttt ctctgcagtg ttacacgaac tttttacaat ctactactac
      121 taacgaacgt cgcttgtatg tgttgcgctc tgaaaagttc acatctgttg ctgccttttg
      181 tacattattt gagaatcttt actcatttcg tgttctttta tcaatttagt gaataaactg
      241 attcatattg attaacaagt agttgaacaa aaagaaacgg tgtccttttg tgtttgcatt
      301 ttatctctgt cctttgtagt ctgattttaa ttttttctta ctctattcat ctccttctct
      361 gctccaccaa accggtatat atgactaggg cttacacaga cgtgcttcat tttcttttct
      421 tgcttcttcc ccacaaatga tacactgtca atgggcgagg aagaagagct gcttcttggg
      481 acggactcgt cgttctttct catcaatcac cggcgatgca ctcgacacaa acacgttaca
      541 agaaagaccc tcccgggctc tttcagccca ttttgttgaa atatgtgctt ttgaaacgca
      601 acattcgaat gatttgtcac gacagtcggc ctaattaatt gactctcgaa aatgagcata
      661 cgattgagtg ttaatgtgcc gaagatgtag ggaggagcaa gaaaagaggg tggaagagat
      721 gatgagtgac agactgtctt aacagatgac ctgcttgtgg tggtcacctg tcgagttatg
      781 atctcagtaa ttgcttcaat tttccacata cgttcccaag tgtctcatta gaatataaat
      841 aagacaatat tttgttctgt gtcctgtaga gaatacacaa ttccaactag aaacgttttt
      901 tttcattttt ttaaattgat ttcaaaacat cttttgacct gatgtcagtg tgggggattt
      961 ttttggcatt tgtaatcgaa attaatgaat ttttagacaa tttctgaaaa atagtaaggt
     1021 ttaaaaaaat cttaaggacg gcacacactg tacaacaata ctttgactga tggtttttct
     1081 gtttttcatt acaatcttat tttttctagt cagctttttt atgtgattat ttaaaaaaac
     1141 aatggtcttg ctttagatag tcaagatctc acactcttaa taaaaaaggt tttaatactt
     1201 cagattttgc tttaactttt ggaaattgcc aaccgtttct gaaaaatttg aagtctttga
     1261 aaacaattac agcacttttg cacgttttgc tgctatcaaa tgtttaatat aaataagcaa
     1321 aatatttgtg aaaaataaaa tacttcactt cattttttaa attttgcttc tttctcacaa
     1381 atgatatatt gactgtaaat gtcagaattt tgaaaagtta aaaaaaatta tgttgaaaat
     1441 ttattttttt ttcttactga cagactgatg tgcctttttc agcatcttga ataagttcaa
     1501 gtttttttaa tcgaagctgt ttccagtgtc tgtcagtgac ccacacacta gaacgcctaa
     1561 aataaaaatc gaaatttttg gagtatccga atttaatttg gaatcaaatt gatgcttgaa
     1621 atatatgtga gctcttaaaa taataataaa aaattgctac aatcatttta ttaaaatatt
     1681 tttcgatcta aaaattgttc atcattgagt aatgtaattt tgataaatta gacccccccc
     1741 ccttcttcta caaaactgat caaaaggtcc gaaagactta gaagacccgc acaatagtag
     1801 aatattcaat ggccgaatag tgaatgcgcg aggcgggtgg ttgctgtctt ctcggttcat
     1861 ataaggctca tctgttcaat caccaaatga tcgatgtgtt tttgtggcct tcggcgtcag
     1921 tttggagcag cacttcaacc tcattgaaac ctctgtcttc ctcctatctt tgcctttttt
     1981 tagtcaagag gcggtgtgaa cgacgcaata ttcccgttgg ttatcaccac actagaagtt
     2041 tttttgagca ctcttatcag ggaacaatta ttgatgattt attgccaact agtgggttgg
     2101 ttttgctatt ttttgaaatt tactggtatg gtcttttgag attttcaggc agttaaaact
     2161 tactaagact gatggttaac aaaaagttgt aatatacaat actaactgaa gcgaaacttt
     2221 gaacgaaaaa tatattaaac tgttaaattt tatttgcagc tagctaacta ataatttgta
     2281 tgtttagtgc atttaaaaaa gttttcagct atattataat gatagtttga aactatgata
     2341 aaactgagtt tatacgactt tttgaataca caaaaaaaat ttaaaaattt tttgaaaact
     2401 tttttaatgt attttttgta gtattttatt ttccctggcc taggaattat gaaaaaaagt
     2461 ttctagattt caatttattc cttattttga caaattatta tgaaataatt cattaatttc
     2521 attttgtacg taaatgggcg ggactaatga gatcctcaaa ctccttgtga ttctctttcc
     2581 tcctcgcctt cttctaccat actcctggac ctcgctcttc ccgcgcactc tcttccccct
     2641 ctttcactct tgccacactt gccaagggag tcataaatcg tgcaacatgt ttagtaggtg
     2701 ccgttttgaa atatagatgc tctttttttc tttctcacaa accatctttt agttggtctc
     2761 gctcatctat tttgaaaaat gacgatccgt gtagagcttt gattcttcat agttattttc
     2821 ttgtgtgtgc gtcgggcctc atactgccag gtagagtgta cgtgtttacc taacaaggga
     2881 gaggtctatg tggcactgcg cacttcatcg tatgcaaatt tgcttttatg atctcttccc
     2941 tcaggtcatt gtccatcata agcatccgca cacttttttt ttggtgtttt cttctctttt
     3001 tttgggtcac cagttcacaa ccaaaaatat aggtggtcaa aacataacat ttttctgtac
     3061 ttacagtaag tggaaatgtc gtctaagcac acttagaata ttctaaataa aagagaatta
     3121 ttagaaaaca actttaaaat gtttctggca acgttttgaa agtttgagaa atttataaca
     3181 aacgttcaaa tcaagttgca acataattag aatataaaat ttgtgaaaaa cactttttgg
     3241 tcaacttcca aaattatcaa tgatgaaaaa gtgactgttt gcaactattt ggctttaaat
     3301 taaagaacta attttttatt ttaattttca ctatttccct acaagcaacc tgtaaagtga
     3361 tattgtgtgt gactattttt tctcttttac cacaatattt gagtattaac ccccctttag
     3421 ctgtcttcaa ttatcattta attcagaatt tcaggtaggc gcgcctctag aggatccccg
     3481 ggattggcca aaggacccaa aggtatgttt cgaatgatac taacataaca tagaacattt
     3541 tcaggaggac ccttggctag cagctctcct gcagggttcg acaaaaatgg tgagcaaggg
     3601 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     3661 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     3721 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
     3781 gacctacggc gtgcagtgct tcgcccgcta ccccgaccac atgaaacagc acgacttctt
     3841 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3901 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3961 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     4021 ctacaacagc cacaaggtct atatcaccgc cgacaagcag aagaacggca tcaaggtgaa
     4081 cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     4141 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     4201 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     4261 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggtaccg gtggatccgg
     4321 aatggcggct ggcggacaac ctcaaggcgc tacaccggga caacccgatc agaactttga
     4381 ctacatgttc aagctcctga taatcggaaa ttcatcagtt ggaaaaacat cattcctctt
     4441 ccgttactgt gatgattcat tcacttctgc cttcgtctct actgtcggaa tcgatttcaa
     4501 agtgaaaact gtgttccgtg gagacaaacg agtcaaactt caaatctggg ataccgccgg
     4561 acaggagagg taccgtacca tcaccaccgc ctactatcgt ggagcaatgg gattcattct
     4621 gatgtatgac atcactaatg aagagtcttt taatagtgtt caggattggt gcactcaaat
     4681 caagacatac tcatgggaaa atgctcaagt tgttttggtt ggaaataaat gtgatatgga
     4741 ctctgaaaga gttgtatcta tggatagggg acgccaactt gctgatcaac ttggtttgga
     4801 attcttcgaa acatcagcca aggagaacat taatgtaaag gcagtttttg agaagttggt
     4861 ggagattatt tgtgataaga tggcagagag tttggataag gacccacagc aacagccaaa
     4921 aggacagaag ctcgaagcga atccgaccca aaagcctgct caacagcaat gcaattgcta
     4981 aatcagatat cactcgagga gctcccgaga tcttcagcgc tcagcattcg tagaattgta
     5041 agtttaaact ttctcatact aactaacgcg ctctatttaa attttcagaa ttccaactga
     5101 gcgccggtcg ctaccattac caacttgtct ggtgtcaaaa ataatagggg ccgctgtcat
     5161 cagagtaagt ttaaactgag ttctactaac taacgagtaa tatttaaatt ttcagcatct
     5221 cgcgcccgtg cctctgactt ctaagtccaa ttactcttca acatccctac atgctctttc
     5281 tccctgtgct cccaccccct atttttgtta ttatcaaaaa acttctctta atttctttgt
     5341 tttttagctt cttttaagtc acctctaaca atgaaattgt gtagattcaa aaatagaatt
     5401 aattcgtaat aaaaagtcga aaaaaattgt gctccctccc cccattaata ataattctat
     5461 cccaaaatct acacaatgtt ctgtgtacac ttcttatgtt ttttacttct gataaatttt
     5521 tttgaaacat catagaaaaa accgcacaca aaatacctta tcatatgtta cgtttcagtt
     5581 tatgaccgca atttttattt cttcgcacgt ctgggcctct catgacgtca aatcatgctc
     5641 atcgtgaaaa agttttggag tatttttgga atttttcaat caagtgaaag tttatgaaat
     5701 taattttcct gcttttgctt tttggggttt cccctattgt ttgtcaagat ttcgaggacg
     5761 gcgtttttct tgctaaaatc acaagtattg atgagcacga tgcaagaaag atcggaagaa
     5821 ggtttgggtt tgaggctcag tggaaggtga gtagaagttg ataatttgaa agtggagtag
     5881 tgtctatggg gtttttgcct taaatgacag aatacattcc caatatacca aacataactg
     5941 tttcctacta gtcggccgta cgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     6001 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     6061 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     6121 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     6181 agatgcgtaa ggagaaaata ccgcatcagg cggccttaat taagggcctc gtgatacgcc
     6241 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6301 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6361 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6421 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6481 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6541 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6601 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6661 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6721 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6781 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6841 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6901 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6961 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7021 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7081 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     7141 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7201 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7261 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7321 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7381 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7441 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7501 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7561 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7621 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7681 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7741 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7801 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7861 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7921 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7981 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8041 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     8101 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     8161 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     8221 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     8281 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     8341 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtgga
//
