LOCUS       sequence-266236-        8692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prgef-1 INS-3::GFP unc-54 3' UTR C.elegans pan-neural expression of 
            INS-3::GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8692)
  AUTHORS   Hung WL, Wang Y, Chitturi J, Zhen M
  TITLE     A Caenorhabditis elegans developmental decision requires insulin 
            signaling-mediated neuron-intestine communication.
  JOURNAL   Development. 2014 Apr;141(8):1767-79. doi: 10.1242/dev.103846. Epub 
            2014 Mar 26.
  PUBMED    24671950
REFERENCE   2  (bases 1 to 8692)
  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: 
            "10.1242/dev.103846"; journalName: "Development"; date: "2014-04"; 
            volume: "141"; issue: "8"; pages: "1767-79"
COMMENT     Sequence Label: pJH2933
FEATURES             Location/Qualifiers
     source          1..8692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      3517..3526
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3523..3525,3526..3528,3529..4242)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3523..3525,3526..3528,3529..4239)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3568..3589)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3829..3848)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4176..4197
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4267..4291)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4317..4358
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(5184..5201)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5184..5200)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5193..5215)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5413..5868
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5500..5519)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5710..5731
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5885..5904)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6004..6026
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6064..6082)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6150..6254
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6255..6323,6324..7115)
                     /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(6473..6492)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7286..7874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7775..7794
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8028..8045
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8162..8183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8198..8203,8204..8221,8222..8228)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8236..8252
                     /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     8241..8263
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8260..8276
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8260..8276
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aggcaaaaag agcggtgcaa aaatataaga gactgcaata ctaatagagg aaatacggta
       61 attgaatttt agtgaaactt gcgacagttt tcctcatttt tttgttattc cggcatacga
      121 gtgtggcata cgagtgaggt catcttttgt ttcttccgtt tcttcatcgc ttttgaaaaa
      181 aaatgttgaa gaaaacttcg tagccgatag ccggataatg tttggcacgg cgttccaaca
      241 tatatcattg gaaatttttt aagatttttg cgaaaaatca cattcttcac gatgagaaca
      301 cgttattgaa ggaatataaa tcaagaataa catatagtta tatttctcta ttactttaac
      361 gttaaatatg agcaaatttg agcattttga ttgcgatgaa aagcagaatc gagtcaactg
      421 aaatccgttc aaaaacctaa gctcggttcg cccgaaagtc gatttttcca atagccgaac
      481 agcactgctt ctttattttc caggtatttt tggggtgtaa gaattatatc ttctttgttt
      541 gggtatttac gtttccgaat tccctttctt gatctgtcgc tatttttcag aaaccagaac
      601 agtttatcct ttttcatgag aacctgaata gctcaaaaac ccgtttaatt tcttttcatc
      661 atctcatttg aaactttcct agtcttctta atgatttcca ggctcctcct acatttgtat
      721 ctcaagatgc atacacttta tttccgctcc aattcgaatc atctacttct cttttttctt
      781 caactttcta cttttttcca ttcttttctt tcaaatgttc gacgtcttca agtgactgct
      841 cttcattctc ctcatttttc tcgaatttca ttcttgtctc ttttttgctc aaaaatggaa
      901 aatgaaagtg acgtgagatt tggacggcgg gacacggggg cagtagaagc agcaaaaagg
      961 agagaaagag gacacaaata agaagaacga attcaaaaat aagcggagag gagctatttc
     1021 cgtcaattct acctccccaa tcttcatcaa ttcggctcaa ttgaatgacg tcacagggag
     1081 ataaaccgtt tggatgagcg ccgtccatca ctgacgccat cccgtttggg acaagaaaag
     1141 agaaaaaaga gcacaaagtt tttggtgacg gatcttgtca atcatatgaa agttgttctg
     1201 attgattgtc agtttttttc ctactttttt ggattctatc cacttctgaa cttttgacaa
     1261 gtttcaaact ttctgaatca ttttctatgc attttcctgg aattcttttt atgtaaaata
     1321 tgaaatagaa tgtttttgaa ttcaagttct gcttttttct tcttttctgt tctgttcggg
     1381 cttgggtatg ctttttttaa aaattatttt gcacatcgac caataagtgc gcaacttata
     1441 aaattaattt atttttgtta atttttgaaa tacttgtatt gctttaagtg atctgacctc
     1501 gccctgagct ttccacgtag ttatcaaata caaatcctcc aagggtaacg tacctatatt
     1561 actgatcttt ataataactt tatcacctgt ccagttccag aggaattctg ttaagcttat
     1621 aggcacagaa ggagtcattt ctgctggttt ttaatgatcc aaatctttat ttcaagtaaa
     1681 aaactgaaca cttgcgaata aaactatcag attaaccatt caccaaaaat gtgtttgaat
     1741 ctaaaacttt ctcagtattc caaatataga aataaataac cacgacattg ctaaaatctg
     1801 tctgaattgt gtactcctta ccgtgaagta ataatggata taatgaatcg tttgaaatga
     1861 atgatcagca catttttggt gaaagatcac aaataaggaa taagcgtcgg aaaataaaac
     1921 gattatttcg gatcaaaaat ttgttgaaga tcatatacac tcgagaccaa gattattcta
     1981 gacaattttc aaattggctt ctttgtttgc aaatcttcat aataatctgt gaagtttgga
     2041 aatttgaatt tttaatcttt tttcatagat tatagttttt atttctttgc aaaactatat
     2101 taaaaaccga tgcattgttt tagggaaatt aatgagcctt ttgttcaaca ctaaaaacaa
     2161 taaaattaaa attttggctt catcatttga ccttttttaa gttcgaaaac ttttctcgta
     2221 tttctgaacc gccaattttt tcacacatct ctagactttt ggtgcccgtt ccagaaagtt
     2281 aagtaattgc tattctaaga aagttctcaa cattgttttt tagttctgat tgaattctga
     2341 tgttccagga atatatttta aatttaatat tctttgcact atttctatac taactaaata
     2401 ataaatagtc taagtatgtt caatgagaac aaaaaagctc tatctatatt tttctatcct
     2461 atttcattta tatccttttc attttgaact caccagcttc tcttcttctt cttgtccata
     2521 tacttcttaa aagtctcgaa cttctctctc tcactctcca tcaatttctt tatggctacc
     2581 gcttgcgctc tgccgcttcc gaagaaagaa ggcgttggca gagctctcaa ttctattttt
     2641 tctcgccgta ggctatcgat ttctcagctt ctcttctctc ttcctcactc caccaccacc
     2701 acttcgggct tcttcttttc ttcattttcg tcttcttctt catcctcttc tttttttttc
     2761 agttccccct cactcctccc cttatatgcg cgtgcgagag gggtgcaaaa gcagcgcgca
     2821 tccgagaatt gaaacgagaa aacggagacg cagcagcagt tcgtcctcag aaaaatagcc
     2881 agtaaaaaga gaaaaagata gagagagacc tatcgcattt tattttcaaa ttgcaattcc
     2941 tgcaattcat gtgtgcgtgt gcccattttc aattcttccc gttatttttc agaccattac
     3001 caacgttctt tctagatctt gagacaatct tccttctgct caatcgttcg tcgtgaagac
     3061 ggcatcgacg acgacgacga tcagcagtaa aggatcctta tgaaactctc cgttgttctt
     3121 gcacttttca ttattttcca acttggagct gcaagtctta tgcgtaactg gatgttcgat
     3181 tttgagaaag aattggaaca cgattatgat gattcggaaa ttggattcca taacattcac
     3241 tccctgatgg ccagatcaag aagaggagac aaagtgaaga tttgtggtac aaaagttctg
     3301 aaaatggtga tggtaatgtg tggaggagaa tgttcatgta agttgtattt gttacaaacg
     3361 tagtaattca aatcttatgg gaaatgtaaa actcattcaa aacgaaaaat attttcagca
     3421 acgaatgaga acatcgctac agaatgctgt gaaaaaatgt gcacaatgga agatataact
     3481 actaagtgct gcccttcaag acgggatcca ccggtcgcca ccatggtgag caagggcgag
     3541 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3601 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     3661 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     3721 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     3781 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3841 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3901 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     3961 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     4021 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     4081 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     4141 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     4201 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cactcgagat
     4261 atctagaccc agctttcttg tacaaagtgg ttgatctaga gggcccgcgg ttcgaaggta
     4321 agcctatccc taaccctctc ctcggtctcg attctacgcg taccggttag taatgagttt
     4381 atctgagctc cgcatcggcc gctgtcatca gatcgccatc tcgcgcccgt gcctctgact
     4441 tctaagtcca attactcttc aacatcccta catgctcttt ctccctgtgc tcccaccccc
     4501 tatttttgtt attatcaaaa aacttctctt aatttctttg ttttttagct tcttttaagt
     4561 cacctctaac aatgaaattg tgtagattca aaaatagaat taattcgtaa taaaaagtcg
     4621 aaaaaaattg tgctccctcc ccccattaat aataattcta tcccaaaatc tacacaatgt
     4681 tctgtgtaca cttcttatgt tttttacttc tgataaattt ttttgaaaca tcatagaaaa
     4741 aaccgcacac aaaatacctt atcatatgtt acgtttcagt ttatgaccgc aatttttatt
     4801 tcttcgcacg tctgggcctc tcatgacgtc aaatcatgct catcgtgaaa aagttttgga
     4861 gtatttttgg aatttttcaa tcaagtgaaa gtttatgaaa ttaattttcc tgcttttgct
     4921 ttttggggtt tcccctattg tttgtcaaga tttcgaggac ggcgtttttc ttgctaaaat
     4981 cacaagtatt gatgagcacg atgcaagaaa gatcggaaga aggtttgggt ttgaggctca
     5041 gtggaaggtg agtagaagtt gataatttga aagtggagta gtgtctatgg ggtttttgcc
     5101 ttaaatgaca gaatacattc ccaatatacc aaacataact gtttctacta gtggccacgt
     5161 gggccgtgca ccttaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     5221 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     5281 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     5341 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     5401 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5461 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5521 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5581 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     5641 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5701 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     5761 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5821 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     5881 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     5941 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     6001 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     6061 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     6121 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6181 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6241 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6301 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6361 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6421 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6481 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6541 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6601 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6661 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6721 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6781 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6841 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6901 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6961 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7021 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7081 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7141 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7201 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7261 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7321 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7381 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7441 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7501 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7561 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7621 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7681 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7741 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7801 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7861 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7921 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7981 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     8041 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     8101 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     8161 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     8221 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga
     8281 ttacgaattc gaatggccat gggacgtcga cgcttgcatg cctgcagcgt ttccgatacc
     8341 cccttatatc agcacacatt cagtcatcat accacaaaat gttgtgatag tgataggcaa
     8401 aactaccgta tttcctctat taatcttgca tgcaagacta attttcgatt aacccgtagg
     8461 ggtgcaagac taatagagac tgcaagacta ttagaggctg aaatactaat tttcgtatgc
     8521 tcaataattt tggaaattgg cctatttttt gtagaaactt gataccgttt aaacaaggaa
     8581 aaatacacac tttttaatat ttcatcaata atttgaacga ttttgtgatt ttaagttcaa
     8641 tttgcccaaa aaaagacaat ttttctgacg ttaccgcaaa gtaatgctgg tc
//
