LOCUS       sequence-37590-e        5983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5983)
  AUTHORS   Hochbaum D, Zhang Y, Stuckenholz C, Labhart P, Alexiadis V, Martin 
            R, Knolker HJ, Fisher AL
  TITLE     DAF-12 Regulates a Connected Network of Genes to Ensure Robust 
            Developmental Decisions.
  JOURNAL   PLoS Genet. 2011 Jul;7(7):e1002179. Epub 2011 Jul 21.
  PUBMED    21814518
REFERENCE   2  (bases 1 to 5983)
  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"; journalName: "PLoS 
            Genet."; date: "2011-07"; volume: "7(7)"; pages: "e1002179. Epub 
            2011 Jul 21"
COMMENT     Sequence Label: pCMV myc DAF-12
FEATURES             Location/Qualifiers
     source          1..5983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     531..555
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          672..768
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     polyA_signal    3122..3256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3146..3165)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3200..3219
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3367..3383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3367..3383)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3380..3402)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3391..3407
                     /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(3415..3421,3422..3439,3440..3445))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3460..3481
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3598..3615)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3769..4357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3849..3868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4528..5319,5320..5388))
                     /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     5151..5170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5389..5493)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5561..5579
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5617..5639)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5739..5758
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5952..5974
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5966..5983
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5967..5983
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat ggcatcaatg
      841 cagaagctga tctcagagga ggacctgctt atggccatgg aggccggcac aaatggagga
      901 gtcattgcgg aacagtcaat ggaaatcgaa acaaatgaaa acccagataa ggtagaggag
      961 cccgtcgtga gaaggaaacg agtaacccgc aggcgccaca ggcggataca cagcaagaat
     1021 aactgcttga cacctccaaa cagcgatgac gaccctcaaa tgagcacacc agatgatcca
     1081 gtcatccaca gtccaccatc aattggagca gctcctggta tgaatgggta tcatggatca
     1141 ggagtgaaac ttgaagaaag ttctggtgct tgtggctctc cggatgatgg cctgttagac
     1201 tccagcgaag agagtcgacg acgtcagaaa acatgtagag tatgtggaga tcatgcgacg
     1261 ggctataatt tcaacgttat cacatgcgaa tcttgtaaag cattcttcag gagaaatgcg
     1321 ttacgtccca aggagttcaa atgtccatat tctgaagatt gcgaaattaa ttctgtaagc
     1381 agacggtttt gtcagaaatg tcgactgagg aaatgcttca cagttggtat gaaaaaagag
     1441 tggattttaa acgaagaaca actgcggcgg aggaaaaact ctcgtttgaa caatacggga
     1501 acgtgtaata aaagatccca gccagggaat cagcagtcac cacaaggtcc taatcaacag
     1561 ccacatttat caccgcatca tccaggagtt gcaatatatc caccacaacc acaacgccca
     1621 ctaacaatca atccaatgga caatcaaatg atgcatcata tgcaagctaa cagaccaaat
     1681 gccatgccac aacttatcag tccaccagga gcacaaccgt atccattgac atccccagtc
     1741 ggctctagtg catctgactc tccaccaaac cgaagtttaa caatgatgca caacggcgaa
     1801 aaaagtccag atggctatga tccaaacatt atggctcatc gtgccccgcc gccatccttc
     1861 aacaataggc ctaaaatgga tagtggacag gttgttcttt ccacggaaga atataaacaa
     1921 ctcttgtctc gaattcccgg tgcccaagtc cctggtctta tgaatgaaga agaacctatc
     1981 aacaaacgtg cggcatacaa ttgcaatggt catccaatgc cagctgaaac aacaccaccc
     2041 tacagtgctc cgatgagtga catgagcctc tcgcgccaca actccacatc atctaatagt
     2101 gcacaaaatc atttcgatat tgcatcgttt ggtatgggaa tcgtgactgc aaccggaggt
     2161 ggtgatgctg ccgaagagat gtacaaacgg atgaacatgt tttacgaaaa ttgcatccaa
     2221 tcggctctcg acagcccaga aaatcaagaa ccaaaaccac aagaggcaat gattccaaag
     2281 gaagaataca tgactccaac acatggtttt caatatcaat ctgatccgta tcaagttcca
     2341 ccagcagaaa gaaacatcaa ttatcaactg aatgccgctg agcttaaagc actggatgct
     2401 gtccgagaag ctttctatgg tatggacgac ccaatggagc agggacgtca aatgcagtca
     2461 ttccttaagg caaacaaaac tccagctgat attatgaata ttatggatgt taccatgagg
     2521 cgtttcgtca aagttgccaa aggtgttccg gcgttccgtg aggtatctca ggaaggaaag
     2581 ttttcgcttc tcaagggggg aatgattgaa atgctgacag tccgtggtgt gacccgctac
     2641 gacgcttcga ccaactcttt caagacacca acaattaagg ggcaaaacgt ttccgtcaac
     2701 gtggatgata tgtttgctaa attgaatgca aatgcacaag ctcagaaagc taaatgttta
     2761 gagttcttcg gtttcttcga cgaggagatt aaaaagaatg aacttgccgt ttatctagtc
     2821 atgttggctg ttctcttctc tgtccgctct gatcctccga tgaacgaaaa cgatgttcgc
     2881 atcgttacag aacgacataa tcatttcatg agtcttctca acagatattt ggaatcgttg
     2941 ttcggagagc aggcccgccg tatctttgaa cggattccaa aagcactggg attacttaat
     3001 gaaattgctc gtaatgctgg aatgctgttc atgggaactg tgcgttctgg tgaagccgaa
     3061 gagctgccag gagaattttt caaaatcaaa tagagatctc tcgaggtacc gcggccgcgg
     3121 ggatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     3181 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     3241 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     3301 ggtgtgggag gttttttcgg atcctctaga gtcgatctgc aggcatgcta gcttggcgta
     3361 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3421 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3481 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3541 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3601 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3661 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3721 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3781 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3841 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3901 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3961 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4021 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4081 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4141 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4201 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4261 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4321 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4381 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4441 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4501 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4561 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4621 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4681 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4741 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4801 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4861 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4921 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4981 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5041 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5101 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5161 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5221 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5281 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5341 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5401 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5461 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5521 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     5581 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     5641 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     5701 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     5761 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     5821 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5881 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     5941 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agt
//
