LOCUS       sequence-266280-        8624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prgef-1 INS-6::GFP unc-54 3' UTR C.elegans pan-neural expression of 
            INS-6::GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8624)
  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 8624)
  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: pJH2256
FEATURES             Location/Qualifiers
     source          1..8624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1202..1211
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1208..1210,1211..1213,1214..1927)
                     /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(1208..1210,1211..1213,1214..1924)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1253..1274)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1514..1533)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1861..1882
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1952..1976)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2002..2043
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2869..2886)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2869..2885)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2878..2900)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3098..3553
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3185..3204)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3395..3416
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3570..3589)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3689..3711
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3749..3767)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3835..3939
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3940..4008,4009..4800)
                     /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(4158..4177)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4971..5559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5460..5479
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5713..5730
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5847..5868
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5883..5888,5889..5906,5907..5913)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5921..5937
                     /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     5926..5948
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5945..5961
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5945..5961
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tctctagact tttggtgccc gttccagaaa gttaagtaat tgctattcta agaaagttct
       61 caacattgtt ttttagttct gattgaattc tgatgttcca ggaatatatt ttaaatttaa
      121 tattctttgc actatttcta tactaactaa ataataaata gtctaagtat gttcaatgag
      181 aacaaaaaag ctctatctat atttttctat cctatttcat ttatatcctt ttcattttga
      241 actcaccagc ttctcttctt cttcttgtcc atatacttct taaaagtctc gaacttctct
      301 ctctcactct ccatcaattt ctttatggct accgcttgcg ctctgccgct tccgaagaaa
      361 gaaggcgttg gcagagctct caattctatt ttttctcgcc gtaggctatc gatttctcag
      421 cttctcttct ctcttcctca ctccaccacc accacttcgg gcttcttctt ttcttcattt
      481 tcgtcttctt cttcatcctc ttcttttttt ttcagttccc cctcactcct ccccttatat
      541 gcgcgtgcga gaggggtgca aaagcagcgc gcatccgaga attgaaacga gaaaacggag
      601 acgcagcagc agttcgtcct cagaaaaata gccagtaaaa agagaaaaag atagagagag
      661 acctatcgca ttttattttc aaattgcaat tcctgcaatt catgtgtgcg tgtgcccatt
      721 ttcaattctt cccgttattt ttcagaccat taccaacgtt ctttctagat cttgagacaa
      781 tcttccttct gctcaatcgt tcgtcgtgaa gacggcatcg acgacgacga cgatcagcag
      841 taaaggatcc ctatgaactc tgtctttact atcatcttcg ttttgtgcgc actccaagtc
      901 gctgcaagtt tccgtcaatc cttcggtcct tcaatgtctg aagaatcagc aagcatgcaa
      961 cttctccgtg aacttcaaca caacatgatg gaatcagctc accgaccaat gccacgagca
     1021 agacgtgttc cagcaccagg agaaactcgt gcctgcggaa gaaaactcat ctctttagtc
     1081 atggctgtct gtggagatct ttgcaaccca caagaaggaa aggacattgc gactgaatgc
     1141 tgcggaaatc agtgttctga tgactacata agatctgctt gttgtccaaa ggatcccggt
     1201 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1261 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1321 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1381 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1441 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1501 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1561 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1621 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1681 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1741 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1801 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1861 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1921 caagtaaagc ggccgcactc gagatatcta gacccagctt tcttgtacaa agtggttgat
     1981 ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg tctcgattct
     2041 acgcgtaccg gttagtaatg agtttatctg agctccgcat cggccgctgt catcagatcg
     2101 ccatctcgcg cccgtgcctc tgacttctaa gtccaattac tcttcaacat ccctacatgc
     2161 tctttctccc tgtgctccca ccccctattt ttgttattat caaaaaactt ctcttaattt
     2221 ctttgttttt tagcttcttt taagtcacct ctaacaatga aattgtgtag attcaaaaat
     2281 agaattaatt cgtaataaaa agtcgaaaaa aattgtgctc cctcccccca ttaataataa
     2341 ttctatccca aaatctacac aatgttctgt gtacacttct tatgtttttt acttctgata
     2401 aatttttttg aaacatcata gaaaaaaccg cacacaaaat accttatcat atgttacgtt
     2461 tcagtttatg accgcaattt ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc
     2521 atgctcatcg tgaaaaagtt ttggagtatt tttggaattt ttcaatcaag tgaaagttta
     2581 tgaaattaat tttcctgctt ttgctttttg gggtttcccc tattgtttgt caagatttcg
     2641 aggacggcgt ttttcttgct aaaatcacaa gtattgatga gcacgatgca agaaagatcg
     2701 gaagaaggtt tgggtttgag gctcagtgga aggtgagtag aagttgataa tttgaaagtg
     2761 gagtagtgtc tatggggttt ttgccttaaa tgacagaata cattcccaat ataccaaaca
     2821 taactgtttc tactagtggc cacgtgggcc gtgcacctta agcttggcac tggccgtcgt
     2881 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     2941 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     3001 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     3061 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3121 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3181 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3241 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3301 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3361 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3421 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     3481 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3541 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3601 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3661 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3721 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3781 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3841 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3901 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3961 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4021 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4081 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4141 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4201 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4261 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4321 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4381 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4441 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4501 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4561 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4621 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4681 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4741 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4801 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4861 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4921 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5041 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5101 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5161 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5221 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5281 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5341 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5401 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5461 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5521 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5581 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5641 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5701 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     5761 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     5821 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     5881 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     5941 cacacaggaa acagctatga catgattacg aattcgaatg gccatgggac gtcgacgctt
     6001 gcatgcctgc agcgtttccg ataccccctt atatcagcac acattcagtc atcataccac
     6061 aaaatgttgt gatagtgata ggcaaaacta ccgtatttcc tctattaatc ttgcatgcaa
     6121 gactaatttt cgattaaccc gtaggggtgc aagactaata gagactgcaa gactattaga
     6181 ggctgaaata ctaattttcg tatgctcaat aattttggaa attggcctat tttttgtaga
     6241 aacttgatac cgtttaaaca aggaaaaata cacacttttt aatatttcat caataatttg
     6301 aacgattttg tgattttaag ttcaatttgc ccaaaaaaag acaatttttc tgacgttacc
     6361 gcaaagtaat gctggtcagg caaaaagagc ggtgcaaaaa tataagagac tgcaatacta
     6421 atagaggaaa tacggtaatt gaattttagt gaaacttgcg acagttttcc tcattttttt
     6481 gttattccgg catacgagtg tggcatacga gtgaggtcat cttttgtttc ttccgtttct
     6541 tcatcgcttt tgaaaaaaaa tgttgaagaa aacttcgtag ccgatagccg gataatgttt
     6601 ggcacggcgt tccaacatat atcattggaa attttttaag atttttgcga aaaatcacat
     6661 tcttcacgat gagaacacgt tattgaagga atataaatca agaataacat atagttatat
     6721 ttctctatta ctttaacgtt aaatatgagc aaatttgagc attttgattg cgatgaaaag
     6781 cagaatcgag tcaactgaaa tccgttcaaa aacctaagct cggttcgccc gaaagtcgat
     6841 ttttccaata gccgaacagc actgcttctt tattttccag gtatttttgg ggtgtaagaa
     6901 ttatatcttc tttgtttggg tatttacgtt tccgaattcc ctttcttgat ctgtcgctat
     6961 ttttcagaaa ccagaacagt ttatcctttt tcatgagaac ctgaatagct caaaaacccg
     7021 tttaatttct tttcatcatc tcatttgaaa ctttcctagt cttcttaatg atttccaggc
     7081 tcctcctaca tttgtatctc aagatgcata cactttattt ccgctccaat tcgaatcatc
     7141 tacttctctt ttttcttcaa ctttctactt ttttccattc ttttctttca aatgttcgac
     7201 gtcttcaagt gactgctctt cattctcctc atttttctcg aatttcattc ttgtctcttt
     7261 tttgctcaaa aatggaaaat gaaagtgacg tgagatttgg acggcgggac acgggggcag
     7321 tagaagcagc aaaaaggaga gaaagaggac acaaataaga agaacgaatt caaaaataag
     7381 cggagaggag ctatttccgt caattctacc tccccaatct tcatcaattc ggctcaattg
     7441 aatgacgtca cagggagata aaccgtttgg atgagcgccg tccatcactg acgccatccc
     7501 gtttgggaca agaaaagaga aaaaagagca caaagttttt ggtgacggat cttgtcaatc
     7561 atatgaaagt tgttctgatt gattgtcagt ttttttccta cttttttgga ttctatccac
     7621 ttctgaactt ttgacaagtt tcaaactttc tgaatcattt tctatgcatt ttcctggaat
     7681 tctttttatg taaaatatga aatagaatgt ttttgaattc aagttctgct tttttcttct
     7741 tttctgttct gttcgggctt gggtatgctt tttttaaaaa ttattttgca catcgaccaa
     7801 taagtgcgca acttataaaa ttaatttatt tttgttaatt tttgaaatac ttgtattgct
     7861 ttaagtgatc tgacctcgcc ctgagctttc cacgtagtta tcaaatacaa atcctccaag
     7921 ggtaacgtac ctatattact gatctttata ataactttat cacctgtcca gttccagagg
     7981 aattctgtta agcttatagg cacagaagga gtcatttctg ctggttttta atgatccaaa
     8041 tctttatttc aagtaaaaaa ctgaacactt gcgaataaaa ctatcagatt aaccattcac
     8101 caaaaatgtg tttgaatcta aaactttctc agtattccaa atatagaaat aaataaccac
     8161 gacattgcta aaatctgtct gaattgtgta ctccttaccg tgaagtaata atggatataa
     8221 tgaatcgttt gaaatgaatg atcagcacat ttttggtgaa agatcacaaa taaggaataa
     8281 gcgtcggaaa ataaaacgat tatttcggat caaaaatttg ttgaagatca tatacactcg
     8341 agaccaagat tattctagac aattttcaaa ttggcttctt tgtttgcaaa tcttcataat
     8401 aatctgtgaa gtttggaaat ttgaattttt aatctttttt catagattat agtttttatt
     8461 tctttgcaaa actatattaa aaaccgatgc attgttttag ggaaattaat gagccttttg
     8521 ttcaacacta aaaacaataa aattaaaatt ttggcttcat catttgacct tttttaagtt
     8581 cgaaaacttt tctcgtattt ctgaaccgcc aattttttca caca
//
