LOCUS       sequence-9275-ex        6086 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 6086)
  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 6086)
  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: pGC90
FEATURES             Location/Qualifiers
     source          1..6086
                     /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             164..184
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             206..226
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             248..268
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             290..310
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(353..381)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1129..1149
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             complement(1791..1802)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3837..3941
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3942..4010,4011..4802)
                     /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(4160..4179)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4973..5561
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5462..5481
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5715..5732
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5849..5870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5885..5890,5891..5908,5909..5915)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5923..5939
                     /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     5928..5950
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5947..5963
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5947..5963
                     /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 ccccgggtac cgagctcaga aaaaatgact gctccaaaga agaagcgtaa
      181 ggtaccggca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg cagaaaaaat
      241 gactgctcca aagaagaagc gtaaggtacc ggcagaaaaa atgactgctc caaagaagaa
      301 gcgtaaggta ccggtagaaa aaatgagtaa aggagaagaa cttttcactg gagttgtccc
      361 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg
      421 tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact
      481 acctgttcca tgggtaagtt taaacatata tatactaact aaccctgatt atttaaattt
      541 tcagccaaca cttgtcacta ctttctgtta tggtgttcaa tgcttctcga gatacccaga
      601 tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag
      661 aactatattt ttcaaagatg acgggaacta caagacacgt aagtttaaac agttcggtac
      721 taactaacca tacatattta aattttcagg tgctgaagtc aagtttgaag gtgataccct
      781 tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca
      841 caaattggaa tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa
      901 tggaatcaaa gttgtaagtt taaacatgat tttactaact aactaatctg atttaaattt
      961 tcagaacttc aaaattagac acaacattga agatggaagc gttcaactag cagaccatta
     1021 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
     1081 cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga
     1141 gtttgtaaca gctgctggga ttacacatgg catggacgaa ctatacaaaa gcccacaagc
     1201 tttggaattc gctagccggc catacaagta atccggatga tcgacgccaa cgtcgttgaa
     1261 ttttcaaatt ttaaatactg aatatttgtt ttttttccta ttatttattt attctctttg
     1321 tgtttttttt cttgctttct aaaaaattaa ttcaatccaa atctaaacat gagcggtttt
     1381 ttttctcttt ccgtctccca attcgtattc cgctcctctc atctgaacac aatgtgcaag
     1441 tttatttatc ttctcgcttt catttcatta ggacgtgggg ggaattggtg gaagggggaa
     1501 acacacaaaa ggatgatgga aatgaaataa ggacacacaa tatgcaacaa cattcaattc
     1561 agaaatatgg aggaaggttt aaaagaaaac ataaaaatat atagaggagg aaggaaaact
     1621 agtctagaat cctatgcttg cactttgagc caatttatcc aagtccttgt aaaaagtatt
     1681 cgaaaattgt taacggataa aatgtttatt ataatatcaa aaacaatttg tcagttgacc
     1741 actttttgat atatttgaca gaaaccggga tgaattggct caaagttagg gcgcccttct
     1801 attacagggt ttctgataac aaaccggtta ttaactccca acaagggatg atttcaattc
     1861 atcatgctca aattgaccca aattaagtta catgacaaat tcatcgccct tttcaactct
     1921 tcttggctca tcatctgtta ttctgttcct attctgtctg caccctatac ccctttgcat
     1981 actctctcgt cattcctttc ttggatagtg tcttcttctc cccagctctg ctacttctat
     2041 gacttgcgcc cgctgctttt ccgctcgttc ctctcctctc tgacgtcttc ttctgcttct
     2101 tttctccttc ctttttccct tctcaatctt cctctttttc ccatttctgt caatcattcg
     2161 aagaagaaga agaagaagac cctcattcat ttcatttttt tttctgtcgg tgtgtgctgc
     2221 tcggttaaga gtgagctctc tattccacgt cttcttcttt ttcttcttcg attcgaatca
     2281 atcactccac caaaacgcat tcgtttttgg gattcacccc gcggttcgca acaggtttct
     2341 ttttcaaatt attagcgtta taaatagaaa aatggtggag tttcaataaa aatgataatt
     2401 tacaaagtgt attttgatta catgtactca aaaaggttga aatttttcat accagttttc
     2461 cggaaatcca tctgatatca ttatcgtatt tttcttttta aaaatgtttt caaaaaaaaa
     2521 caaaatatag ctggttattt ggcaccctct aattaccatt ttcctgtcac accacactct
     2581 tttccttctt ccacttcttt ccgttttcag ccgcttccaa ccaaaccgat atgaaagccg
     2641 agcaacaaca atcgattcca cccggctcgg cgaccttccc gtcgcaggtg agactcagaa
     2701 aactagagaa accgctcaac taactcttga tatccgattt cattcttttc tcttttcttt
     2761 ttgttgaact tttccatatt tccagatgcc acggccacca ccaagcaccg aacaaggaat
     2821 cacaacggaa tcggagcttg cgaagaaagc tcaaatcact ccgaacgacg ttttagcact
     2881 tccgggaatc actcaaggta tcttgctccc tttggatttc agtttaaaac ataaatttag
     2941 gattcttatg ctccccatct gcgaacatct ataacatcga gttcaccaag ttccaaatcc
     3001 gtgatctgga cactgagcaa gtgcttttcg agatcgccaa accggagaac gatcaggaga
     3061 atgatgagtc gccacaggag tcggcaagat acgtgcgtta tagatttgct ccaaactttt
     3121 tgaaactcaa aacggtcgga gcaactgtgg aattcaaagt aggagacatc ccaatccatc
     3181 atttccgaat gatcgaacgt cacttcttca aagatcgcct tctgaagtgt tttgactttg
     3241 aattcggatt ctgtattccg aattcacgaa acaactgtga acatatctat gagttccctc
     3301 aactctctca acaactcagt gagtcattat tctaaaaagt acaattcaaa agactaatct
     3361 ctttttcagt ggacgacatg atcaacaatc caaacgagac tcgttctgac agcttctatt
     3421 tcgtcgataa caaactcgtc atgcacaaca aagccgacta ctcatatgat gcataaatat
     3481 ttaatacaaa aatgttctgg ataattattc tgtcgaatag aaaaaaaact ccaaatgtga
     3541 ttaaattcca ataattcctg tctagtttgt tccttcccct tccccttctc atgttcaatg
     3601 cattcctaag cttttcagtt ccccccttgt ttctatattt tttcgctgtc ctgtcacact
     3661 cgctaaaaac actaatcaca cggaaatctg ttttcaataa aaactccaac tttaactcat
     3721 tttcaatttc aactgaaaga ttttttcatt agagaatgtc taggcatgct ctagagggcc
     3781 caaaaggcct tttttttttg gcgccgacgt caggtggcac ttttcgggga aatgtgcgcg
     3841 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3901 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3961 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4021 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4081 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4141 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4201 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4261 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4321 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4381 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4441 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4501 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4561 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4621 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4681 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     4741 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     4801 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     4861 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4921 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4981 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5041 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5101 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5161 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5221 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5281 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5341 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     5401 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5461 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5521 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5581 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5641 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5701 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     5761 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     5821 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     5881 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     5941 ttcacacagg aaacagctat gaccatgatt acgccaagct gtaagtttaa acatgatctt
     6001 actaactaac tattctcatt taaattttca gagcttaaaa atggctgaaa tcactcacaa
     6061 cgatggatac gctaacaact tggaaa
//
