LOCUS       sequence-346666-        9931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pmyo-3 GFP::DAF-16a unc-54 3' UTR C.elegans muscle expression of 
            isoforms a of DAF-16.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9931)
  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 9931)
  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: pJH2972
FEATURES             Location/Qualifiers
     source          1..9931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     34..51
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    168..189
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(204..209,210..227,228..234)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    242..258
                     /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     247..269
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     266..282
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     266..282
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2784..2812)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3560..3580
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(7121..7138)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7121..7137)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7130..7152)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7350..7805
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7437..7456)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7647..7668
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7822..7841)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7941..7963
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8001..8019)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8087..8191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8192..8260,8261..9052)
                     /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(8410..8429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9223..9811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9712..9731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
       61 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      121 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      181 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      241 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattcgaat
      301 ggccatggga cgtcgacgcg gccgccggct ataataagtt cttgaataaa ataattttcc
      361 cgacaaaaca tgagtatttc tttcgaaaat aaaagtgcag gctaattaga gattattctg
      421 taattaactg cataatttgt cacgtgccat agttttacat tccactacgt catagttctt
      481 aaaatactaa tctcctgaaa atagaagtag gtgaagaaag tttaattatc agttctaaaa
      541 tgacaattga tctttggaat atgttctgaa actaccgatc attgaacaga tgctatttga
      601 atgatataga attgtatatt tgcaatttct gaaacgcgtt cttaaaggca cacagattaa
      661 ttcaaaaggg tctggccgca aaaaggttta tggtggccga ttttgagttt tgtgtgtgat
      721 tgctttttca caatcagtgt tttcaggatt atgtgatgaa ctagatcttc aagtttcgtt
      781 acatttcata tgttttcgga actcacgaag tacatattgg gtattgtgct caaaaaattc
      841 agcaatcagc ttcgctccgc tgactttaga acccaaaaaa atagtatggc caaactgact
      901 gtgttacgat catttcaatt tttcaataca tatttaagat ttctaagagt aagaaggtca
      961 aaaactgttc tggaatacat atatattttt caggttacaa ttagtcaaaa agtgcactga
     1021 aatatacgtt ttaatttcac gaataaccca attagttcaa tgtatttttg gtcaaccaac
     1081 gttaaagttt ggcttccaac caattatcat ttctgatcaa ccacaatgtt ttttctttat
     1141 ctgcaagtta attttttatt tttatccaga tgtttggcat atttttcaat tcttcactag
     1201 cgcccacttc ttgcacttcc ggcgccctga atctaatgca tctgttgcaa gaattgaaag
     1261 accaatcaac acattgtttt cttcacgaga tactgaagaa aatgaataaa aacagagaaa
     1321 aagagccatg tgattagtga caactgttgc taacagatac catagcttgg acttggtacg
     1381 tgatggcaac gtatgggtca acaaaaatga ttgcagaggg ggtgcaaaac agtcaagtcg
     1441 agaaaatatg aaaaacagaa aacaaagaac agaaaaatgg gtttgagagt cagtataatt
     1501 tataaaagaa aaattgtaca tagaaattaa ccatttttgt agaagaagtt atttttcaag
     1561 catcgttaaa aattattcaa agcaccttat ttcatattta attttaaaca tggttaaatg
     1621 aacaacacgg tgcgcaatca ggaaaacttg aaatctgaaa ctgttgttgt gatcttcttc
     1681 gcaactgttc agatagcact agtgtaatgt taagagtgcg cgaatataat ggaatataat
     1741 ggatcacacc tcctgccatc aggtaaacgt ctctgttatc acatatttcc aactattaaa
     1801 tttttacctt ttacagtttt acattttttt gaaaaaagta actttttgtc ttcaaaatcc
     1861 ctgacgaaaa tatcaaatat tttaatcgag actgcagagg aaccgattga tgatttggaa
     1921 aatccagctt tacctgtgta agaactgaaa agtttcataa ccctagggta ttcccagtta
     1981 cattccccac tggctaacaa tagcacccag tttttcatca cctttcttca aatttctcgg
     2041 cgatttgtta aaaacaaaat ttgtgtccct tctctgatat ctctatgtct ctaaacacaa
     2101 gttcatcgga aaacgaagga gggtaggtgt tggttgggct cccgaagtga aaatagaaga
     2161 gcaagaatag aatattagag agagagtgca gagagggcgg gatagctccc gggattccgt
     2221 tttcttcttc tttatcttca acgatgatgt gtgtgcgtgt tgtatagatt ctgttgctcc
     2281 cccacaactc gctccgaagg ctcaatacaa ttcaattgat attggaggag agcctaccgg
     2341 agtgggagga taagaagaaa cataagaaga agaagaagaa gaagcatgct tctggttttt
     2401 gatgctatga aaacggcaca aaaagatgat tgaggtccct tttcaatacc ttctctcatc
     2461 tttcaaatcc cattgaaacc taaaacttct caccacgctt taccattgtt ctccaaaaac
     2521 ttatagcaat gtctataact tttttatctc tgaaaagcag tgttccattt ttctttttcc
     2581 tattttattt caattgtttc tcacatttcg tttggattct ttgcttgtca accagcttct
     2641 tcttccactt ttaccgtcta attttcaggg cagggagcca tcaaacccac gaccactaga
     2701 tccatctagg gatccccggg taccgagctc gaattcaagt accggtagaa aaaatgagta
     2761 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
     2821 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
     2881 cccttaaatt tatttgcact actggaaaac tacctgttcc atgggtaagt ttaaacatat
     2941 atatactaac taaccctgat tatttaaatt ttcagccaac acttgtcact actttctgtt
     3001 atggtgttca atgcttctcg agatacccag atcatatgaa acggcatgac tttttcaaga
     3061 gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact
     3121 acaagacacg taagtttaaa cagttcggta ctaactaacc atacatattt aaattttcag
     3181 gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg
     3241 attttaaaga agatggaaac attcttggac acaaattgga atacaactat aactcacaca
     3301 atgtatacat catggcagac aaacaaaaga atggaatcaa agttgtaagt ttaaacatga
     3361 ttttactaac taactaatct gatttaaatt ttcagaactt caaaattaga cacaacattg
     3421 aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc
     3481 ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca
     3541 acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg
     3601 gcatggacga actatacaaa agcccacaag cttacatgtc ctgggcccat cgattcggac
     3661 gtcccgaggg cttaagcgga cgcgtcgagg ggatccctat gatggagatg ctggtagatc
     3721 agggaactga tgcatcgtca tccgcctcca cgtccacctc atctgtttcg agattcggag
     3781 cggacacgtt catgaataca ccggatgatg tgatgatgaa tgatgatatg gaaccgattc
     3841 ctcgtgatcg gtgcaatacg tggccaatgc gtaggccgca actcgaacca ccactcaact
     3901 cgagtcccat tattcatgaa caaattcctg aagaagatgc tgacctatac gggagcaatg
     3961 agcaatgtgg acagctcggc ggagcatctt caaacgggtc gacagcaatg cttcatactc
     4021 cagatggaag caattctcat cagacatcgt ttccttcgga aatgtccgaa tcgccagacg
     4081 ataccgtatc gggaaaaaag acaacgacca gacggaacgc ttggggaaat atgtcatatg
     4141 ctgaacttat cactacagcc attatggcta gtccagagaa acggttaact cttgcacaag
     4201 tttacgaatg gatggtccag aatgttccat acttcaggga taagggagat tcgaacagtt
     4261 cagctggatg gaagaactcg atccgtcaca atctgtctct tcattctcgt ttcatgcgaa
     4321 ttcagaatga aggagccgga aagagctcgt ggtgggttat taatccagat gcaaagccag
     4381 gaaggaatcc acggcgtaca cgtgaacgat ccaatactat tgagacgact acaaaggctc
     4441 aactcgaaaa atctcgccgc ggagccaaga agaggataaa ggagagagca ttgatgggct
     4501 cccttcactc gacacttaat ggaaattcga ttgccggatc gattcaaacg atttctcacg
     4561 atttgtatga tgatgattca atgcaaggag catttgataa cgttccatcg tctttccggt
     4621 tagtaattta gttttatttt tattttttga attttccaaa tttccaaatt ttttcagtcc
     4681 ccgaactcaa tcgaacctct cgattcctgg atcgtcgtct cgtgtttctc cagctattgg
     4741 aagtgatatc tatgatgatc tagaattccc atcatcggtt ggcgaatcgg ttccagcaat
     4801 tccaagtgat attgttgata gaactgatca aatgcgtatc gatgcaacta ctcatattgg
     4861 tggagttcag attaagcagg agtcgaagcc gattaaggtg agaaatgaga atttgcaatt
     4921 ttcaggaaaa tttgaaaatt ttcttgtggt tagtacaagt gagctccttt ttagacaatt
     4981 aaaaaatttg aattttactt aaattcaata tttttaattc tgaatttagt ttcaaggtcc
     5041 cctaaaatat tagtaacatg caaatcattt tatattttct cacaccgaaa attagaattt
     5101 tctttttttt taagtgaaaa aactgagtaa tatatagaat tttcagacgg aaccaattgc
     5161 tccaccacca tcataccacg agttgaacag tgtccgtgga tcgtgtgctc agaatccact
     5221 tcttcgaaat ccaattgtgc caagcactaa cttcaagccg ttagtttttt tttgaacaga
     5281 agttaattaa tctatcgtaa tttcacaaga ttttgggtcc cggcgctcaa aaatgttttt
     5341 tttaaatttt tatgttaaaa atagttttaa atcagttttg aaagtgtaaa ttttggctta
     5401 aaattatttt aaacagaaaa ttggaaataa aacgaagaaa taaacttttt tgcgcgccga
     5461 gacccatctg aataaatgcg tgcgccttta agaaacctgt gaaattacta tatatacaaa
     5521 attctgagaa tgcgtattac gcaacaaatt tgacgcgcaa aatatctcgt agcgaaaact
     5581 acagtaatcc ttaaaatgag tactgtagcg cttgcgtcga tttacgggct cgattttcgt
     5641 gaattaattt attttcgaaa agtgacagcg atattacatt taactttgtt ctttctaatc
     5701 attttctcat ttttgtgaat ttttaaaaat aatttttcga ttaataaagg attgccgtaa
     5761 atcgacacat ggtgctacag taggcattta aaggattact gtagttttcg ctacgagata
     5821 tttcgcgcgt caaatatgtt gcgtaatacg cattctcaga attttatgat cccataatat
     5881 atatttcttc tttaattcaa tatataataa ttatccaaaa tttgttcaga atgccactac
     5941 cgggtgccta tggaaactat caaaatggtg gaataactcc aatcaattgg ctatcaacat
     6001 ccagctcatc tccactgcct ggaattcaat cgtgtggaat tgtagctgca cagcatactg
     6061 tcgcttcttc atcggctctt ccaattgatt tggaaaatct gacacttccc gatcagccac
     6121 tgatggatac tatggatgtt gatgcattga tcagacatga gctgagtcaa gctggagggc
     6181 agcagcatat tcattttgat ttgtaactgc aggaattcga tatgcagaag ctgatctcag
     6241 aggaggacct gcttcagaag cttatctcag aggaggacct tcgacctcga gggggggccc
     6301 ggtaccatgg tattgatatc tgagctccgc atcggccgct gtcatcagat cgccatctcg
     6361 cgcccgtgcc tctgacttct aagtccaatt actcttcaac atccctacat gctctttctc
     6421 cctgtgctcc caccccctat ttttgttatt atcaaaaaac ttctcttaat ttctttgttt
     6481 tttagcttct tttaagtcac ctctaacaat gaaattgtgt agattcaaaa atagaattaa
     6541 ttcgtaataa aaagtcgaaa aaaattgtgc tccctccccc cattaataat aattctatcc
     6601 caaaatctac acaatgttct gtgtacactt cttatgtttt ttacttctga taaatttttt
     6661 tgaaacatca tagaaaaaac cgcacacaaa ataccttatc atatgttacg tttcagttta
     6721 tgaccgcaat ttttatttct tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat
     6781 cgtgaaaaag ttttggagta tttttggaat ttttcaatca agtgaaagtt tatgaaatta
     6841 attttcctgc ttttgctttt tggggtttcc cctattgttt gtcaagattt cgaggacggc
     6901 gtttttcttg ctaaaatcac aagtattgat gagcacgatg caagaaagat cggaagaagg
     6961 tttgggtttg aggctcagtg gaaggtgagt agaagttgat aatttgaaag tggagtagtg
     7021 tctatggggt ttttgcctta aatgacagaa tacattccca atataccaaa cataactgtt
     7081 tctactagtg gccacgtggg ccgtgcacct taagcttggc actggccgtc gttttacaac
     7141 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     7201 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     7261 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     7321 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     7381 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     7441 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     7501 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     7561 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     7621 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     7681 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     7741 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     7801 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     7861 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     7921 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     7981 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     8041 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     8101 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     8161 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     8221 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     8281 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     8341 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     8401 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     8461 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     8521 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     8581 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     8641 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     8701 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     8761 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     8821 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     8881 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     8941 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     9001 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     9061 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     9121 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     9181 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     9241 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     9301 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     9361 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     9421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     9481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     9541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     9601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     9661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     9721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     9781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     9841 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     9901 ggataaccgt attaccgcct ttgagtgagc t
//
