LOCUS       sequence-9274-ex        5939 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 5939)
  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 5939)
  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: pGC89
FEATURES             Location/Qualifiers
     source          1..5939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(57..104)
                     /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             143..163
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(206..234)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     982..1002
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(1644..1655)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3690..3794
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3795..3863,3864..4655)
                     /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(4013..4032)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4826..5414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5315..5334
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5568..5585
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5702..5723
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5738..5743,5744..5761,5762..5768)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5776..5792
                     /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     5781..5803
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5800..5816
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5800..5816
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcagcc cgggggatcc actagttcta gcttcaaaag cgctctgaag
       61 ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaagatc cccgggtacc
      121 gagctcagaa aaaatgacgg cacctaaaaa gaaacgcaaa gtaccggtag aaaaaatgag
      181 taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
      241 taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact
      301 tacccttaaa tttatttgca ctactggaaa actacctgtt ccatgggtaa gtttaaacat
      361 atatatacta actaaccctg attatttaaa ttttcagcca acacttgtca ctactttctg
      421 ttatggtgtt caatgcttct cgagataccc agatcatatg aaacggcatg actttttcaa
      481 gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa
      541 ctacaagaca cgtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc
      601 aggtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat
      661 tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca
      721 caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttgtaa gtttaaacat
      781 gattttacta actaactaat ctgatttaaa ttttcagaac ttcaaaatta gacacaacat
      841 tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg
      901 ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc
      961 caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca
     1021 tggcatggac gaactataca aaagcccaca agctttggaa ttcgctagcc ggccatacaa
     1081 gtaatccgga tgatcgacgc caacgtcgtt gaattttcaa attttaaata ctgaatattt
     1141 gttttttttc ctattattta tttattctct ttgtgttttt tttcttgctt tctaaaaaat
     1201 taattcaatc caaatctaaa catgagcggt tttttttctc tttccgtctc ccaattcgta
     1261 ttccgctcct ctcatctgaa cacaatgtgc aagtttattt atcttctcgc tttcatttca
     1321 ttaggacgtg gggggaattg gtggaagggg gaaacacaca aaaggatgat ggaaatgaaa
     1381 taaggacaca caatatgcaa caacattcaa ttcagaaata tggaggaagg tttaaaagaa
     1441 aacataaaaa tatatagagg aggaaggaaa actagtctag aatcctatgc ttgcactttg
     1501 agccaattta tccaagtcct tgtaaaaagt attcgaaaat tgttaacgga taaaatgttt
     1561 attataatat caaaaacaat ttgtcagttg accacttttt gatatatttg acagaaaccg
     1621 ggatgaattg gctcaaagtt agggcgccct tctattacag ggtttctgat aacaaaccgg
     1681 ttattaactc ccaacaaggg atgatttcaa ttcatcatgc tcaaattgac ccaaattaag
     1741 ttacatgaca aattcatcgc ccttttcaac tcttcttggc tcatcatctg ttattctgtt
     1801 cctattctgt ctgcacccta tacccctttg catactctct cgtcattcct ttcttggata
     1861 gtgtcttctt ctccccagct ctgctacttc tatgacttgc gcccgctgct tttccgctcg
     1921 ttcctctcct ctctgacgtc ttcttctgct tcttttctcc ttcctttttc ccttctcaat
     1981 cttcctcttt ttcccatttc tgtcaatcat tcgaagaaga agaagaagaa gaccctcatt
     2041 catttcattt ttttttctgt cggtgtgtgc tgctcggtta agagtgagct ctctattcca
     2101 cgtcttcttc tttttcttct tcgattcgaa tcaatcactc caccaaaacg cattcgtttt
     2161 tgggattcac cccgcggttc gcaacaggtt tctttttcaa attattagcg ttataaatag
     2221 aaaaatggtg gagtttcaat aaaaatgata atttacaaag tgtattttga ttacatgtac
     2281 tcaaaaaggt tgaaattttt cataccagtt ttccggaaat ccatctgata tcattatcgt
     2341 atttttcttt ttaaaaatgt tttcaaaaaa aaacaaaata tagctggtta tttggcaccc
     2401 tctaattacc attttcctgt cacaccacac tcttttcctt cttccacttc tttccgtttt
     2461 cagccgcttc caaccaaacc gatatgaaag ccgagcaaca acaatcgatt ccacccggct
     2521 cggcgacctt cccgtcgcag gtgagactca gaaaactaga gaaaccgctc aactaactct
     2581 tgatatccga tttcattctt ttctcttttc tttttgttga acttttccat atttccagat
     2641 gccacggcca ccaccaagca ccgaacaagg aatcacaacg gaatcggagc ttgcgaagaa
     2701 agctcaaatc actccgaacg acgttttagc acttccggga atcactcaag gtatcttgct
     2761 ccctttggat ttcagtttaa aacataaatt taggattctt atgctcccca tctgcgaaca
     2821 tctataacat cgagttcacc aagttccaaa tccgtgatct ggacactgag caagtgcttt
     2881 tcgagatcgc caaaccggag aacgatcagg agaatgatga gtcgccacag gagtcggcaa
     2941 gatacgtgcg ttatagattt gctccaaact ttttgaaact caaaacggtc ggagcaactg
     3001 tggaattcaa agtaggagac atcccaatcc atcatttccg aatgatcgaa cgtcacttct
     3061 tcaaagatcg ccttctgaag tgttttgact ttgaattcgg attctgtatt ccgaattcac
     3121 gaaacaactg tgaacatatc tatgagttcc ctcaactctc tcaacaactc agtgagtcat
     3181 tattctaaaa agtacaattc aaaagactaa tctctttttc agtggacgac atgatcaaca
     3241 atccaaacga gactcgttct gacagcttct atttcgtcga taacaaactc gtcatgcaca
     3301 acaaagccga ctactcatat gatgcataaa tatttaatac aaaaatgttc tggataatta
     3361 ttctgtcgaa tagaaaaaaa actccaaatg tgattaaatt ccaataattc ctgtctagtt
     3421 tgttccttcc ccttcccctt ctcatgttca atgcattcct aagcttttca gttcccccct
     3481 tgtttctata ttttttcgct gtcctgtcac actcgctaaa aacactaatc acacggaaat
     3541 ctgttttcaa taaaaactcc aactttaact cattttcaat ttcaactgaa agattttttc
     3601 attagagaat gtctaggcat gctctagagg gcccaaaagg cctttttttt ttggcgccga
     3661 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3721 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3781 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3841 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3901 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3961 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4021 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4081 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4141 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4201 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4261 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4321 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4381 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4441 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4501 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4561 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4621 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4681 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4741 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4801 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4861 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4921 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4981 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5041 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5101 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5161 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt
     5221 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5281 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5341 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5401 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5461 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5521 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5581 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5641 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5701 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5761 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     5821 attacgccaa gctgtaagtt taaacatgat cttactaact aactattctc atttaaattt
     5881 tcagagctta aaaatggctg aaatcactca caacgatgga tacgctaaca acttggaaa
//
