LOCUS       sequence-9283-ex        4833 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 4833)
  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 4833)
  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: pGC296
FEATURES             Location/Qualifiers
     source          1..4833
                     /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"
     CDS             370..1047
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFL"
     CDS             1090..1764
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQA
                     LKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHL
                     FL"
     primer_bind     complement(2273..2295)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2395..2414
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2492..2510)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2578..2682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2683..2751,2752..3543)
                     /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(2901..2920)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3714..4302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4203..4222
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4456..4473
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4590..4611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4626..4631,4632..4649,4650..4656)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4664..4680
                     /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     4669..4691
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4688..4704
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4688..4704
                     /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 ggtagacgat
      361 aaggatccga tggtggcctc ctccgaggac gtcatcaaag agttcatgcg cttcaaggtg
      421 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      481 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
      541 gcctgggaca tcctgtcccc ccagttccag tacggctcca aggcgtacgt gaagcacccc
      601 gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
      661 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
      721 acgctgatct acaaggtgaa gttccgcggc accaacttcc cccccgacgg ccccgtaatg
      781 cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg
      841 ctgaagggcg agatccacca ggccctgaag ctgaaggacg gcggccacta cctggtggag
      901 ttcaagacca tctacatggc caagaagccc gtgcagctgc ccggctacta ctacgtggac
      961 accaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc
     1021 tccgagggcc gccaccacct gttcctgggg catggcaccg gcagcaccgg cagcggcagc
     1081 tccggcaccg cctcctccga ggacgtcatc aaagagttca tgcgcttcaa ggtgcgcatg
     1141 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1201 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
     1261 gacatcctgt ccccccagtt ccagtacggc tccaaggcgt acgtgaagca ccccgccgac
     1321 atccccgact acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     1381 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcacgctg
     1441 atctacaagg tgaagttccg cggcaccaac ttcccccccg acggccccgt aatgcagaag
     1501 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag
     1561 ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag
     1621 accatctaca tggccaagaa gcccgtgcag ctgcccggct actactacgt ggacaccaag
     1681 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag
     1741 ggccgccacc acctgttcct gtaggaattc gctagccggc catacaagta atccggatga
     1801 tcgacgccaa cgtcgttgaa ttttcaaatt ttaaatactg aatatttgtt ttttttccta
     1861 ttatttattt attctctttg tgtttttttt cttgctttct aaaaaattaa ttcaatccaa
     1921 atctaaacat gagcggtttt ttttctcttt ccgtctccca attcgtattc cgctcctctc
     1981 atctgaacac aatgtgcaag tttatttatc ttctcgcttt catttcatta ggacgtgggg
     2041 ggaattggtg gaagggggaa acacacaaaa ggatgatgga aatgaaataa ggacacacaa
     2101 tatgcaacaa cattcaattc agaaatatgg aggaaggttt aaaagaaaac ataaaaatat
     2161 atagaggagg aaggaaaact agtaaaaaat aagcaaagaa attaggcgaa cgatgagaat
     2221 tgtcctcgct tgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     2281 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     2341 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     2401 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     2461 ggagaaaata ccgcatcagg cggccttaag ggcctcgtga tacgcctatt tttataggtt
     2521 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2581 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2641 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2701 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2761 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2821 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2881 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2941 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3001 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3061 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3121 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3181 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3241 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3301 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3361 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3421 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3481 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3541 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3601 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3661 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3721 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3781 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3841 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3901 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3961 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4021 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4081 gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag
     4141 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4201 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4261 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4321 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4381 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4441 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     4501 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     4561 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     4621 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     4681 tttcacacag gaaacagcta tgaccatgat tacgccaagc tgtaagttta aacatgatct
     4741 tactaactaa ctattctcat ttaaattttc agagcttaaa aatggctgaa atcactcaca
     4801 acgatggata cgctaacaac ttggaaatga aat
//
