LOCUS       sequence-212471-        8045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses INS-22-Emerald in the cholinergic motor neuron DA9 for 
            imaging Dense Core 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 8045)
  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 8045)
  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#920
FEATURES             Location/Qualifiers
     source          1..8045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(3145..3147,3148..3150,3151..3864)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3190..3211)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3451..3470)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3798..3819
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4884..4906)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5006..5025
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5107..5125)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5193..5297
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5298..5366,5367..6158)
                     /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(5516..5535)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6329..6917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6818..6837
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7071..7088
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7205..7226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7241..7246,7247..7264,7265..7271)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7279..7295
                     /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     7284..7306
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7303..7319
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7303..7319
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tgtcgagtta tgatctcagt aattgcttca attttccaca tacgttccca agtgtctcat
       61 tagaatataa ataagacaat attttgttct gtgtcctgta gagaatacac aattccaact
      121 agaaacgttt tttttcattt ttttaaattg atttcaaaac atcttttgac ctgatgtcag
      181 tgtgggggat ttttttggca tttgtaatcg aaattaatga atttttagac aatttctgaa
      241 aaatagtaag gtttaaaaaa atcttaagga cggcacacac tgtacaacaa tactttgact
      301 gatggttttt ctgtttttca ttacaatctt attttttcta gtcagctttt ttatgtgatt
      361 atttaaaaaa acaatggtct tgctttagat agtcaagatc tcacactctt aataaaaaag
      421 gttttaatac ttcagatttt gctttaactt ttggaaattg ccaaccgttt ctgaaaaatt
      481 tgaagtcttt gaaaacaatt acagcacttt tgcacgtttt gctgctatca aatgtttaat
      541 ataaataagc aaaatatttg tgaaaaataa aatacttcac ttcatttttt aaattttgct
      601 tctttctcac aaatgatata ttgactgtaa atgtcagaat tttgaaaagt taaaaaaaat
      661 tatgttgaaa atttattttt ttttcttact gacagactga tgtgcctttt tcagcatctt
      721 gaataagttc aagttttttt aatcgaagct gtttccagtg tctgtcagtg acccacacac
      781 tagaacgcct aaaataaaaa tcgaaatttt tggagtatcc gaatttaatt tggaatcaaa
      841 ttgatgcttg aaatatatgt gagctcttaa aataataata aaaaattgct acaatcattt
      901 tattaaaata tttttcgatc taaaaattgt tcatcattga gtaatgtaat tttgataaat
      961 tagacccccc ccccttcttc tacaaaactg atcaaaaggt ccgaaagact tagaagaccc
     1021 gcacaatagt agaatattca atggccgaat agtgaatgcg cgaggcgggt ggttgctgtc
     1081 ttctcggttc atataaggct catctgttca atcaccaaat gatcgatgtg tttttgtggc
     1141 cttcggcgtc agtttggagc agcacttcaa cctcattgaa acctctgtct tcctcctatc
     1201 tttgcctttt tttagtcaag aggcggtgtg aacgacgcaa tattcccgtt ggttatcacc
     1261 acactagaag tttttttgag cactcttatc agggaacaat tattgatgat ttattgccaa
     1321 ctagtgggtt ggttttgcta ttttttgaaa tttactggta tggtcttttg agattttcag
     1381 gcagttaaaa cttactaaga ctgatggtta acaaaaagtt gtaatataca atactaactg
     1441 aagcgaaact ttgaacgaaa aatatattaa actgttaaat tttatttgca gctagctaac
     1501 taataatttg tatgtttagt gcatttaaaa aagttttcag ctatattata atgatagttt
     1561 gaaactatga taaaactgag tttatacgac tttttgaata cacaaaaaaa atttaaaaat
     1621 tttttgaaaa cttttttaat gtattttttg tagtatttta ttttccctgg cctaggaatt
     1681 atgaaaaaaa gtttctagat ttcaatttat tccttatttt gacaaattat tatgaaataa
     1741 ttcattaatt tcattttgta cgtaaatggg cgggactaat gagatcctca aactccttgt
     1801 gattctcttt cctcctcgcc ttcttctacc atactcctgg acctcgctct tcccgcgcac
     1861 tctcttcccc ctctttcact cttgccacac ttgccaaggg agtcataaat cgtgcaacat
     1921 gtttagtagg tgccgttttg aaatatagat gctctttttt tctttctcac aaaccatctt
     1981 ttagttggtc tcgctcatct attttgaaaa atgacgatcc gtgtagagct ttgattcttc
     2041 atagttattt tcttgtgtgt gcgtcgggcc tcatactgcc aggtagagtg tacgtgttta
     2101 cctaacaagg gagaggtcta tgtggcactg cgcacttcat cgtatgcaaa tttgctttta
     2161 tgatctcttc cctcaggtca ttgtccatca taagcatccg cacacttttt ttttggtgtt
     2221 ttcttctctt tttttgggtc accagttcac aaccaaaaat ataggtggtc aaaacataac
     2281 atttttctgt acttacagta agtggaaatg tcgtctaagc acacttagaa tattctaaat
     2341 aaaagagaat tattagaaaa caactttaaa atgtttctgg caacgttttg aaagtttgag
     2401 aaatttataa caaacgttca aatcaagttg caacataatt agaatataaa atttgtgaaa
     2461 aacacttttt ggtcaacttc caaaattatc aatgatgaaa aagtgactgt ttgcaactat
     2521 ttggctttaa attaaagaac taatttttta ttttaatttt cactatttcc ctacaagcaa
     2581 cctgtaaagt gatattgtgt gtgactattt tttctctttt accacaatat ttgagtatta
     2641 accccccttt agctgtcttc aattatcatt taattcagaa tttcaggtag gcgcgcctct
     2701 agaggatccc cgggattggc caaaggaccc aaaggtatgt ttcgaatgat actaacataa
     2761 catagaacat tttcaggagg acccttggcg gccgcgctag cagctctcct gcaaaaaatg
     2821 cacactacaa ctattctcat atgctttttc atctttcttg ttcaagtctc cacaatggat
     2881 gctcacactg acaaatacgt cagaactctg tgtggaaaaa ctgcaatcag aaatattgcc
     2941 aacctttgcc cgccaaagcc agaaatgaag ggtatctgtt ctaccggagg tgagataaac
     3001 ttaaactcaa aagccttgtt tgcaaataat tatttttcta aattttcaga gtatccaagc
     3061 atcaccgaat actgttccat gggattttca gactctcaga tcaagtttat gtgctgtgat
     3121 aaccaagggt tcgacggtac cggtatggtg agcaagggcg aggagctgtt caccggggtg
     3181 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3241 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3301 aagctgcccg tgccctggcc caccctcgtg accaccttga cctacggcgt gcagtgcttc
     3361 gcccgctacc ccgaccacat gaaacagcac gacttcttca agtccgccat gcccgaaggc
     3421 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3481 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3541 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caaggtctat
     3601 atcaccgccg acaagcagaa gaacggcatc aaggtgaact tcaagacccg ccacaacatc
     3661 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3721 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3781 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3841 ggcatggacg agctgtacaa gtaacagata tcactcgagg agctcccgag atcttcagcg
     3901 ctcagcattc gtagaattgt aagtttaaac tttctcatac taactaacgc gctctattta
     3961 aattttcaga attccaactg agcgccggtc gctaccatta ccaacttgtc tggtgtcaaa
     4021 aataataggg gccgctgtca tcagagtaag tttaaactga gttctactaa ctaacgagta
     4081 atatttaaat tttcagcatc tcgcgcccgt gcctctgact tctaagtcca attactcttc
     4141 aacatcccta catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa
     4201 aacttctctt aatttctttg ttttttagct tcttttaagt cacctctaac aatgaaattg
     4261 tgtagattca aaaatagaat taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc
     4321 ccccattaat aataattcta tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt
     4381 tttttacttc tgataaattt ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt
     4441 atcatatgtt acgtttcagt ttatgaccgc aatttttatt tcttcgcacg tctgggcctc
     4501 tcatgacgtc aaatcatgct catcgtgaaa aagttttgga gtatttttgg aatttttcaa
     4561 tcaagtgaaa gtttatgaaa ttaattttcc tgcttttgct ttttggggtt tcccctattg
     4621 tttgtcaaga tttcgaggac ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg
     4681 atgcaagaaa gatcggaaga aggtttgggt ttgaggctca gtggaaggtg agtagaagtt
     4741 gataatttga aagtggagta gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc
     4801 ccaatatacc aaacataact gtttcctact agtcggccgt acgggccctt tcgtctcgcg
     4861 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4921 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4981 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     5041 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa
     5101 ttaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5161 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5221 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5281 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5341 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5401 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5461 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5521 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5581 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5641 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5701 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5761 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5821 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5881 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5941 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6001 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6061 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6121 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6181 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6241 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6301 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6361 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6421 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6481 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6541 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6601 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6661 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6721 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6781 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6841 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6901 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6961 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7021 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     7081 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     7141 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     7201 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     7261 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     7321 atgattacgc caagcttgca tgcctgacac taagtttctc tgcagtgtta cacgaacttt
     7381 ttacaatcta ctactactaa cgaacgtcgc ttgtatgtgt tgcgctctga aaagttcaca
     7441 tctgttgctg ccttttgtac attatttgag aatctttact catttcgtgt tcttttatca
     7501 atttagtgaa taaactgatt catattgatt aacaagtagt tgaacaaaaa gaaacggtgt
     7561 ccttttgtgt ttgcatttta tctctgtcct ttgtagtctg attttaattt tttcttactc
     7621 tattcatctc cttctctgct ccaccaaacc ggtatatatg actagggctt acacagacgt
     7681 gcttcatttt cttttcttgc ttcttcccca caaatgatac actgtcaatg ggcgaggaag
     7741 aagagctgct tcttgggacg gactcgtcgt tctttctcat caatcaccgg cgatgcactc
     7801 gacacaaaca cgttacaaga aagaccctcc cgggctcttt cagcccattt tgttgaaata
     7861 tgtgcttttg aaacgcaaca ttcgaatgat ttgtcacgac agtcggccta attaattgac
     7921 tctcgaaaat gagcatacga ttgagtgtta atgtgccgaa gatgtaggga ggagcaagaa
     7981 aagagggtgg aagagatgat gagtgacaga ctgtcttaac agatgacctg cttgtggtgg
     8041 tcacc
//
