LOCUS       sequence-9284-ex        4707 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 4707)
  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 4707)
  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: pGC309
FEATURES             Location/Qualifiers
     source          1..4707
                     /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..921
                     /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             964..1638
                     /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(2147..2169)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2269..2288
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2366..2384)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2452..2556
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2557..2625,2626..3417)
                     /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(2775..2794)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3588..4176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4077..4096
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4330..4347
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4464..4485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4500..4505,4506..4523,4524..4530)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4538..4554
                     /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     4543..4565
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4562..4578
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4562..4578
                     /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 taccggtaga cgataaggat
      241 ccgatggtgg cctcctccga ggacgtcatc aaagagttca tgcgcttcaa ggtgcgcatg
      301 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
      361 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
      421 gacatcctgt ccccccagtt ccagtacggc tccaaggcgt acgtgaagca ccccgccgac
      481 atccccgact acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
      541 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcacgctg
      601 atctacaagg tgaagttccg cggcaccaac ttcccccccg acggccccgt aatgcagaag
      661 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag
      721 ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag
      781 accatctaca tggccaagaa gcccgtgcag ctgcccggct actactacgt ggacaccaag
      841 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag
      901 ggccgccacc acctgttcct ggggcatggc accggcagca ccggcagcgg cagctccggc
      961 accgcctcct ccgaggacgt catcaaagag ttcatgcgct tcaaggtgcg catggagggc
     1021 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     1081 acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
     1141 ctgtcccccc agttccagta cggctccaag gcgtacgtga agcaccccgc cgacatcccc
     1201 gactacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     1261 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac
     1321 aaggtgaagt tccgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc
     1381 atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag
     1441 atccaccagg ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc
     1501 tacatggcca agaagcccgt gcagctgccc ggctactact acgtggacac caagctggac
     1561 atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc
     1621 caccacctgt tcctgtagga attcgctagc cggccataca agtaatccgg atgatcgacg
     1681 ccaacgtcgt tgaattttca aattttaaat actgaatatt tgtttttttt cctattattt
     1741 atttattctc tttgtgtttt ttttcttgct ttctaaaaaa ttaattcaat ccaaatctaa
     1801 acatgagcgg ttttttttct ctttccgtct cccaattcgt attccgctcc tctcatctga
     1861 acacaatgtg caagtttatt tatcttctcg ctttcatttc attaggacgt ggggggaatt
     1921 ggtggaaggg ggaaacacac aaaaggatga tggaaatgaa ataaggacac acaatatgca
     1981 acaacattca attcagaaat atggaggaag gtttaaaaga aaacataaaa atatatagag
     2041 gaggaaggaa aactagtaaa aaataagcaa agaaattagg cgaacgatga gaattgtcct
     2101 cgcttgggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2161 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2221 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     2281 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2341 aataccgcat caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc
     2401 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2461 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2521 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2581 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2641 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2701 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2761 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2821 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2881 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2941 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3001 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3061 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3121 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3181 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3241 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3301 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3361 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3421 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3481 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3541 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3601 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3661 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3721 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3781 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3841 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3901 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3961 agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4021 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4081 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4141 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4201 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4261 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4321 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     4381 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     4441 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     4501 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     4561 acaggaaaca gctatgacca tgattacgcc aagctgtaag tttaaacatg atcttactaa
     4621 ctaactattc tcatttaaat tttcagagct taaaaatggc tgaaatcact cacaacgatg
     4681 gatacgctaa caacttggaa atgaaat
//
