LOCUS       sequence-9282-ex        4832 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 4832)
  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 4832)
  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: pGC232
FEATURES             Location/Qualifiers
     source          1..4832
                     /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             201..221
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             243..263
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             285..305
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             327..347
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             369..1046
                     /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             1089..1763
                     /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(2272..2294)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2394..2413
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2491..2509)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2577..2681
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2682..2750,2751..3542)
                     /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(2900..2919)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3713..4301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4202..4221
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4455..4472
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4589..4610
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4625..4630,4631..4648,4649..4655)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4663..4679
                     /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     4668..4690
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4687..4703
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4687..4703
                     /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 ccccgggtat gtttcgaatg atactaacat aacatagaac attttcagga
      181 gctcagaaaa aatgactgct ccaaagaaga agcgtaaggt accggcagaa aaaatgactg
      241 ctccaaagaa gaagcgtaag gtaccggcag aaaaaatgac tgctccaaag aagaagcgta
      301 aggtaccggc agaaaaaatg actgctccaa agaagaagcg taaggtaccg gtagacgata
      361 aggatccgat ggtggcctcc tccgaggacg tcatcaaaga gttcatgcgc ttcaaggtgc
      421 gcatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
      481 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg
      541 cctgggacat cctgtccccc cagttccagt acggctccaa ggcgtacgtg aagcaccccg
      601 ccgacatccc cgactacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
      661 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggca
      721 cgctgatcta caaggtgaag ttccgcggca ccaacttccc ccccgacggc cccgtaatgc
      781 agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc
      841 tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt
      901 tcaagaccat ctacatggcc aagaagcccg tgcagctgcc cggctactac tacgtggaca
      961 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct
     1021 ccgagggccg ccaccacctg ttcctggggc atggcaccgg cagcaccggc agcggcagct
     1081 ccggcaccgc ctcctccgag gacgtcatca aagagttcat gcgcttcaag gtgcgcatgg
     1141 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     1201 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg
     1261 acatcctgtc cccccagttc cagtacggct ccaaggcgta cgtgaagcac cccgccgaca
     1321 tccccgacta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     1381 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga
     1441 tctacaaggt gaagttccgc ggcaccaact tcccccccga cggccccgta atgcagaaga
     1501 agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg
     1561 gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga
     1621 ccatctacat ggccaagaag cccgtgcagc tgcccggcta ctactacgtg gacaccaagc
     1681 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg
     1741 gccgccacca cctgttcctg taggaattcg ctagccggcc atacaagtaa tccggatgat
     1801 cgacgccaac gtcgttgaat tttcaaattt taaatactga atatttgttt tttttcctat
     1861 tatttattta ttctctttgt gttttttttc ttgctttcta aaaaattaat tcaatccaaa
     1921 tctaaacatg agcggttttt tttctctttc cgtctcccaa ttcgtattcc gctcctctca
     1981 tctgaacaca atgtgcaagt ttatttatct tctcgctttc atttcattag gacgtggggg
     2041 gaattggtgg aagggggaaa cacacaaaag gatgatggaa atgaaataag gacacacaat
     2101 atgcaacaac attcaattca gaaatatgga ggaaggttta aaagaaaaca taaaaatata
     2161 tagaggagga aggaaaacta gtaaaaaata agcaaagaaa ttaggcgaac gatgagaatt
     2221 gtcctcgctt gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2281 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2341 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2401 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     2461 gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat acgcctattt ttataggtta
     2521 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2581 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2641 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2701 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2761 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2821 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2881 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2941 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3001 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3061 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3121 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3181 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3241 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3301 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3361 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3421 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3481 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3541 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3601 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3661 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3721 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3781 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3841 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3901 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3961 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4021 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4081 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     4141 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4201 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4261 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4321 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4381 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4441 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     4501 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     4561 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     4621 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     4681 ttcacacagg aaacagctat gaccatgatt acgccaagct gtaagtttaa acatgatctt
     4741 actaactaac tattctcatt taaattttca gagcttaaaa atggctgaaa tcactcacaa
     4801 cgatggatac gctaacaact tggaaatgaa at
//
