LOCUS       sequence-9289-ex        4121 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 4121)
  AUTHORS   Voutev R, Hubbard EJ
  TITLE     A ""FLP-Out"" system for controlled gene expression in 
            Caenorhabditis elegans.
  JOURNAL   Genetics. 2008 Sep;180(1):103-19. doi: 10.1534/genetics.108.090274.
  PUBMED    18723890
REFERENCE   2  (bases 1 to 4121)
  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: 
            "10.1534/genetics.108"; journalName: "Genetics"; date: "2008-09"; 
            volume: "180"; issue: "1"; pages: "103-19"
COMMENT     Sequence Label: pGC293
FEATURES             Location/Qualifiers
     source          1..4121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(78..125)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             202..222
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             244..264
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             286..306
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             328..348
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      355..364
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             361..1038
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(541..561)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     953..976
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1561..1583)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1683..1702
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1780..1798)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1866..1970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1971..2039,2040..2831)
                     /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(2189..2208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3002..3590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3491..3510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3744..3761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3878..3899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3914..3919,3920..3937,3938..3944)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3952..3968
                     /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     3957..3979
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3976..3992
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3976..3992
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ccgcggtttt ggcgcggatc cactagttct
       61 agcttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga
      121 acttcaagat ccccggggta tgtttcgaat gatactaaca taacatagaa cattttcagg
      181 agctcagaaa aaatgactgc tccaaagaag aagcgtaagg taccggcaga aaaaatgact
      241 gctccaaaga agaagcgtaa ggtaccggca gaaaaaatga ctgctccaaa gaagaagcgt
      301 aaggtaccgg cagaaaaaat gactgctcca aagaagaagc gtaaggtacc ggtcgccacc
      361 atggcctcct ccgagaacgt catcaccgag ttcatgcgct tcaaggtgcg catggagggc
      421 accgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
      481 cacaacaccg tgaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
      541 ctgtcccccc agttccagta cggctccaag gtgtacgtga agcaccccgc cgacatcccc
      601 gactacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
      661 gacggcggcg tggcgaccgt gacccaggac tcctccctgc aggacggctg cttcatctac
      721 aaggtgaagt tcatcggcgt gaacttcccc tccgacggcc ccgtgatgca gaagaagacc
      781 atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag
      841 acccacaagg ccctgaagct gaaggacggc ggccactacc tggtggagtt caagtccatc
      901 tacatggcca agaagcccgt gcagctgccc ggctactact acgtggacgc caagctggac
      961 atcacctccc acaacgagga ctacaccatc gtggagcagt acgagcgcac cgagggccgc
     1021 caccacctgt tcctgtagcg gccgcgactc tagaattcgc tagccggcca tacaagtaat
     1081 ccggatgatc gacgccaacg tcgttgaatt ttcaaatttt aaatactgaa tatttgtttt
     1141 ttttcctatt atttatttat tctctttgtg ttttttttct tgctttctaa aaaattaatt
     1201 caatccaaat ctaaacatga gcggtttttt ttctctttcc gtctcccaat tcgtattccg
     1261 ctcctctcat ctgaacacaa tgtgcaagtt tatttatctt ctcgctttca tttcattagg
     1321 acgtgggggg aattggtgga agggggaaac acacaaaagg atgatggaaa tgaaataagg
     1381 acacacaata tgcaacaaca ttcaattcag aaatatggag gaaggtttaa aagaaaacat
     1441 aaaaatatat agaggaggaa ggaaaactag taaaaaataa gcaaagaaat taggcgaacg
     1501 atgagaattg tcctcgcttg ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     1561 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1621 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     1681 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     1741 atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg cctcgtgata cgcctatttt
     1801 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     1861 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1921 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1981 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2041 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2101 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2161 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2221 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2281 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2341 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2401 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2461 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2521 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2581 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2641 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2701 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2761 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2821 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2881 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2941 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3001 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3061 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3121 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3361 cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag
     3421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     3781 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     3841 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     3901 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     3961 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctg taagtttaaa
     4021 catgatctta ctaactaact attctcattt aaattttcag agcttaaaaa tggctgaaat
     4081 cactcacaac gatggatacg ctaacaactt ggaaatgaaa t
//
