LOCUS       sequence-128265-        9483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  distal tip cell expression of DAF-16 in C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9483)
  AUTHORS   Michaelson D, Korta DZ, Capua Y, Hubbard EJ
  TITLE     Insulin signaling promotes germline proliferation in C. elegans.
  JOURNAL   Development. 2010 Feb;137(4):671-80. doi: 10.1242/dev.042523.
  PUBMED    20110332
REFERENCE   2  (bases 1 to 9483)
  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"; 
            journalName: "Development"; date: "2010-02"; volume: "137"; issue: 
            "4"; pages: "671-80"
COMMENT     Sequence Label: pGC461
FEATURES             Location/Qualifiers
     source          1..9483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     669..685
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5540..5568)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6316..6336
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(7295..7315)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7295..7313)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7334..7350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7334..7350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7347..7369)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7358..7374
                     /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)."
     promoter        complement(join(7382..7388,7389..7406,7407..7412))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7427..7448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7565..7582)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7736..8324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7816..7835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8495..9286,9287..9355))
                     /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     9118..9137
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9356..9460)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc aggtcgacta
      721 gaggatccgc gacattgtag gaatcatgcg acatttcttt aaaattcccc aaaagctcgt
      781 atttttcaaa attttggcaa tttgccaaaa ctttgaccgg aaattatgaa aaatctaaaa
      841 ttttttggag ttttttatta tgatattcgg ttgttttgga tcattataag tgcatttaga
      901 caaaatcccc actggcgcta ctccaccttt aaattgcagt tgaatttttt ttaaattctc
      961 atttttggaa aatttggaaa tgcaccaaaa ttttgactga aaattatgaa acatttgcaa
     1021 tactttgaaa tttgtgtgat tcatttattt aggacagtta ttagtttacc ttaaatttcc
     1081 ctgaactact ctactccact ttttccacca aattattatt cacgaaccca aaactatctt
     1141 acacatctta tcttacaggt tatacattct gccacgtctt gtaaccccct cccacctatt
     1201 ctctaatata caacctccct ttcaaacagt ccgttatgac tctattgctc ctaatccccc
     1261 ttttgtttcc ggttcggcct gaaacaagaa taacaacagg tgactcgata actgttagga
     1321 aaataaattt tttggtccca atatttcggt atccatttca aaacatcttt taaaattgtc
     1381 aagacagaca attacttctg cttgaaatga aggacacatg tcccctcaat tcctcataaa
     1441 acaaatagag gggtgtcgaa atgataaatg ttgcaagtga ggtgcagatg gcaatgaaat
     1501 ttatatatgt ttctttcatt ccattcttgt ggggcaattt caaacgtttg ataactcaca
     1561 gtggtctaga tttgcagaac atctgtgtta ggtcaagcag agcgagaatt tgagcatttg
     1621 aactttaggg tcctgggggg gggagggggg gggggggggg ggggtgtcta gacagtcagc
     1681 ggcccataag ggttcatatg gtttgacggt gcattgaact attttttaag gaaaaaattg
     1741 agataaaggc catttttcca agaaagcatc tagctttttc aactgacact acaaatttcc
     1801 ctataacttt ctgaaattat cagtgaaaag ttgcccaaat ttgagacttt agctaaaaat
     1861 agacaatttt ttccaaaact ttgagcggcc ataacttttt tttgaaaaac tttcggaacg
     1921 tctcattaca aaatttggtt gctttgagcc attttgaatc cgaaaaagtc tcaaaaagca
     1981 aagtcttaaa tttcggtact ccacctttaa aaaaaatagg gtttccgccg aaatttctgg
     2041 gttccaataa tatctgttac catttccttg ctctgcgacg agcagccacc agacaaaaat
     2101 ctcctaatac ccctcttcat gcctttgttc ttccggtttg gcggtctgaa aaagagcacc
     2161 cctcccggcg gggggctctc ggtatctgtt ccgcaaaaac cgtcatatgg agggctatcg
     2221 ggagacggga agagttggaa atacgggaaa tggtgatgaa taggtagatc aatcgattcc
     2281 ggaacaaata atttgagttg agcacgcgtg ctccacatgt gctcagccgc ttaggtgtga
     2341 gtgaagatcc ttggatttca aggccatcag ccatttcccc tgtccagcta gagcagtcct
     2401 aaacggaatt ttttgcattc ttttgcttct ttacaatttc aaatgttcac tctaactaat
     2461 tcaaaaatct ttctttttta cgagctctcc ttgctcgccg ttgtattcca ttcatacctt
     2521 taagaccacc cgagtgtacg catatttaac caggcaccct aaaaacatga atagtctgtg
     2581 gtggaggtag acagtgtgtt ttctaagaat gtctccgtaa tttgaaatat attttagagt
     2641 cacctgaaat tcatttttca agcaatattt cttaccatat ctttggggat aattttaaaa
     2701 gtggagtgaa gtcgtttagg gacacgcttt taagttttag gaaatggttt ttttttaatt
     2761 cataatttaa aaaaatctat aaaaattgtt acatttttta tgattttctc aattttgttg
     2821 aatgtttgaa tttttttttt gagatgtatt gtttgaattg gagctttttt gtgagttttt
     2881 ttttaatttt gcaattaaat aaaaagtagc taaattagtt ttcgaattcc cgctcaaata
     2941 aatgtcccag aatttacatt tttgagtttt ccggctaaat ttttgccgtt ttttgaatat
     3001 gatttgtaat atttggtctg aaccatggtt tttccaattt tcagaatctt tctcgcaaaa
     3061 aatctccgta tcccgcaaaa taattgttga agcgtcaaaa attggcctaa aagtcgataa
     3121 attaacattg atttcgcttt ttttttagga aaaaagtcta aaagtatttt ttaaactata
     3181 caaaatcatt aaattttgac aagaaaacgc ttaaattaca atttttttta ttttttaaaa
     3241 atttaaaaaa attttaaaaa aacattaaaa ttcagataaa ttgcacaatt tttattgcac
     3301 aagtcgttcg tcgttcgtgt ttttttcgct gtcaaaatgt tggcaaagat tgtgaagtcc
     3361 cctgtagttt aacactctaa gttactccaa agacttctac ctgcctacct gcctgcctat
     3421 ctataaccta gtgcctatca cctactcgct gcatagtttg atgtacctat ataacagttc
     3481 ataaatgaat tttgtcaaaa ttccactctc aagtattctt acacgtactt attttgtaca
     3541 aatccttgtc agtcgctgca agaacataca catcgaagag gcgcaatcga aacaacaggt
     3601 gttcgcccgt tgatcgtctc cccgccccgc ctgttgggcg ggacgggtgt cggtcaccac
     3661 caccacatca tattgttgga cacactttgc acatccggtt cacacccgat taccgcatcg
     3721 gggtcttgat ctggggcggc tattggatct tttgtttatg tagatttttt tctcgccgtt
     3781 atgattttgg attatttttt ctcttatctt gcagtcgact tgtaatcttt gcctgattgc
     3841 tagctagcca aaactttcac ttgttctttt ttttcctcta agtattttcc cgaacttata
     3901 gtcgaccttg agctttgcta gaagccaaca acacttttca aatttgcccg cggatccccg
     3961 ggggatcccc atgatggaga tgctggtaga tcagggaact gatgcatcgt catccgcctc
     4021 cacgtccacc tcatctgttt cgagattcgg agcggacacg ttcatgaata caccggatga
     4081 tgtgatgatg aatgatgata tggaaccgat tcctcgtgat cggtgcaata cgtggccaat
     4141 gcgtaggccg caactcgaac caccactcaa ctcgagtccc attattcatg aacaaattcc
     4201 tgaagaagat gctgacctat acgggagcaa tgagcaatgt ggacagctcg gcggagcatc
     4261 ttcaaacggg tcgacagcaa tgcttcatac tccagatgga agcaattctc atcagacatc
     4321 gtttccttcg gaaatgtccg aatcgccaga cgataccgta tcgggaaaaa agacaacgac
     4381 cagacggaac gcttggggaa atatgtcata tgctgaactt atcactacag ccattatggc
     4441 tagtccagag aaacggttaa ctcttgcaca agtttacgaa tggatggtcc agaatgttcc
     4501 atacttcagg gataagggag attcgaacag ttcagctgga tggaagaact cgatccgtca
     4561 caatctgtct cttcattctc gtttcatgcg aattcagaat gaaggagccg gaaagagctc
     4621 gtggtgggtt attaatccag atgcaaagcc aggaaggaat ccacggcgta cacgtgaacg
     4681 atccaatact attgagacga ctacaaaggc tcaactcgaa aaatctcgcc gcggagccaa
     4741 gaagaggata aaggagagag cattgatggg ctcccttcac tcgacactta atggaaattc
     4801 gattgccgga tcgattcaaa cgatttctca cgatttgtat gatgatgatt caatgcaagg
     4861 agcatttgat aacgttccat catctttccg tccccgaact caatcgaacc tctcgattcc
     4921 tggatcgtcg tctcgtgttt ctccagctat tggaagtgat atctatgatg atctagaatt
     4981 cccatcatgg gttggcgaat cggttccagc aattccaagt gatattgttg atagaactga
     5041 tcaaatgcgt atcgatgcaa ctactcatat tggtggagtt cagattaagc aggagtcgaa
     5101 gccgattaag acggaaccaa ttgctccacc accatcatac cacgagttga acagtgtccg
     5161 tggatcgtgt gctcagaatc cacttcttcg aaatccaatt gtgccaagca ctaacttcaa
     5221 gccaatgcca ctaccgggtg cctatggaaa ctatcaaaat ggtggaataa ctccaatcaa
     5281 ttggctatca acatccaact catctccact gcctggaatt caatcgtgtg gaattgtagc
     5341 tgcacagcat actgtcgctt cttcatcggc tcttccaatt gatttggaaa atctgacact
     5401 tcccgatcag ccactgatgg atactatgga tgttgatgca ttgatcagac atgagctgag
     5461 tcaagctgga gggcagcata ttcattttga tttgccaccg gtagaaaaaa tgagtaaagg
     5521 agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg
     5581 gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct
     5641 taaatttatt tgcactactg gaaaactacc tgttccatgg gtaagtttaa acatatatat
     5701 actaactaac cctgattatt taaattttca gccaacactt gtcactactt tctgttatgg
     5761 tgttcaatgc ttctcgagat acccagatca tatgaaacgg catgactttt tcaagagtgc
     5821 catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa
     5881 gacacgtaag tttaaacagt tcggtactaa ctaaccatac atatttaaat tttcaggtgc
     5941 tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag gtattgattt
     6001 taaagaagat ggaaacattc ttggacacaa attggaatac aactataact cacacaatgt
     6061 atacatcatg gcagacaaac aaaagaatgg aatcaaagtt gtaagtttaa acatgatttt
     6121 actaactaac taatctgatt taaattttca gaacttcaaa attagacaca acattgaaga
     6181 tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg atggccctgt
     6241 ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag atcccaacga
     6301 aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat
     6361 ggatgaacta tacaaatagc attcgtagaa ttccaactga gcgccggtcg ctaccattac
     6421 caacttgtct ggtgtcaaaa ataatagggg ccgctgtcat cagagtaagt ttaaactgag
     6481 ttctactaac taacgagtaa tatttaaatt ttcagcatct cgcgcccgtg cctctgactt
     6541 ctaagtccaa ttactcttca acatccctac atgctctttc tccctgtgct cccaccccct
     6601 atttttgtta ttatcaaaaa acttctctta atttctttgt tttttagctt cttttaagtc
     6661 acctctaaca atgaaattgt gtagattcaa aaatagaatt aattcgtaat aaaaagtcga
     6721 aaaaaattgt gctccctccc cccattaata ataattctat cccaaaatct acacaatgtt
     6781 ctgtgtacac ttcttatgtt ttttacttct gataaatttt tttgaaacat catagaaaaa
     6841 accgcacaca aaatacctta tcatatgtta cgtttcagtt tatgaccgca atttttattt
     6901 cttcgcacgt ctgggcctct catgacgtca aatcatgctc atcgtgaaaa agttttggag
     6961 tatttttgga atttttcaat caagtgaaag tttatgaaat taattttcct gcttttgctt
     7021 tttggggttt cccctattgt ttgtcaagat ttcgaggacg gcgtttttct tgctaaaatc
     7081 acaagtattg atgagcacga tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag
     7141 tggaaggtga gtagaagttg ataatttgaa agtggagtag tgtctatggg gtttttgcct
     7201 taaatgacag aatacattcc caatatacca aacataactg tttcctacta gttctagagc
     7261 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     7321 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     7381 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     7441 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7501 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7561 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7621 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7681 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7741 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7801 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7861 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7921 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     7981 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     8041 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     8101 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     8161 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     8221 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     8281 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     8341 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     8401 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     8461 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     8521 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8581 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8641 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8701 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8761 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8821 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8881 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8941 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     9001 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     9061 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     9121 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     9181 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     9241 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     9301 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     9361 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     9421 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     9481 cac
//
