LOCUS       sequence-9280-ex        4332 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4332)
  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 4332)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pGC80
FEATURES             Location/Qualifiers
     source          1..4332
                     /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             154..831
                     /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             874..1548
                     /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"
     promoter        2077..2181
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2182..2250,2251..3042)
                     /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(2400..2419)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3213..3801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3702..3721
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3955..3972
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4089..4110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4125..4130,4131..4148,4149..4155)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4163..4179
                     /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     4168..4190
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4187..4203
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4187..4203
                     /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 ccccggtaga cgataaggat ccgatggtgg cctcctccga ggacgtcatc
      181 aaagagttca tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca cgagttcgag
      241 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
      301 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc
      361 tccaaggcgt acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc
      421 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
      481 caggactcct ccctgcagga cggcacgctg atctacaagg tgaagttccg cggcaccaac
      541 ttcccccccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag
      601 cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc accaggccct gaagctgaag
      661 gacggcggcc actacctggt ggagttcaag accatctaca tggccaagaa gcccgtgcag
      721 ctgcccggct actactacgt ggacaccaag ctggacatca cctcccacaa cgaggactac
      781 accatcgtgg aacagtacga gcgctccgag ggccgccacc acctgttcct ggggcatggc
      841 accggcagca ccggcagcgg cagctccggc accgcctcct ccgaggacgt catcaaagag
      901 ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg gccacgagtt cgagatcgag
      961 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     1021 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttccagta cggctccaag
     1081 gcgtacgtga agcaccccgc cgacatcccc gactacaaga agctgtcctt ccccgagggc
     1141 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     1201 tcctccctgc aggacggcac gctgatctac aaggtgaagt tccgcggcac caacttcccc
     1261 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg
     1321 tacccccgcg acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc
     1381 ggccactacc tggtggagtt caagaccatc tacatggcca agaagcccgt gcagctgccc
     1441 ggctactact acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc
     1501 gtggaacagt acgagcgctc cgagggccgc caccacctgt tcctgtagga attcgctagc
     1561 cggccataca agtaatccgg atgatcgacg ccaacgtcgt tgaattttca aattttaaat
     1621 actgaatatt tgtttttttt cctattattt atttattctc tttgtgtttt ttttcttgct
     1681 ttctaaaaaa ttaattcaat ccaaatctaa acatgagcgg ttttttttct ctttccgtct
     1741 cccaattcgt attccgctcc tctcatctga acacaatgtg caagtttatt tatcttctcg
     1801 ctttcatttc attaggacgt ggggggaatt ggtggaaggg ggaaacacac aaaaggatga
     1861 tggaaatgaa ataaggacac acaatatgca acaacattca attcagaaat atggaggaag
     1921 gtttaaaaga aaacataaaa atatatagag gaggaaggaa aactagtaaa aaataagcaa
     1981 agaaattagg cgaacgatga gaattgtcct cgcttgggcc caaaaggcct tttttttttg
     2041 gcgccgacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2101 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2161 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2221 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2281 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2341 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2401 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     2461 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2521 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2581 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2641 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2701 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     2761 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2821 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2881 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2941 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3001 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3061 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3121 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3181 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3241 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3301 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3361 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3421 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3481 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3541 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3601 gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3661 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3721 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3781 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3841 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3901 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3961 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4021 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4081 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     4141 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     4201 gaccatgatt acgccaagct gtaagtttaa acatgatctt actaactaac tattctcatt
     4261 taaattttca gagcttaaaa atggctgaaa tcactcacaa cgatggatac gctaacaact
     4321 tggaaatgaa at
//
