LOCUS       sequence-9293-ex        4362 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 4362)
  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 4362)
  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: pGC294
FEATURES             Location/Qualifiers
     source          1..4362
                     /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..264
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             286..306
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             328..348
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(391..419)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1167..1187
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1802..1824)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1924..1943
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2021..2039)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2107..2211
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2212..2280,2281..3072)
                     /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(2430..2449)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3243..3831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3732..3751
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3985..4002
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4119..4140
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4155..4160,4161..4178,4179..4185)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4193..4209
                     /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     4198..4220
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4217..4233
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4217..4233
                     /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 taccggcaga aaaaatgact
      241 gctccaaaga agaagcgtaa ggtaccggca gaaaaaatga ctgctccaaa gaagaagcgt
      301 aaggtaccgg cagaaaaaat gactgctcca aagaagaagc gtaaggtacc ggtagaaaaa
      361 atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
      421 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
      481 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg ggtaagttta
      541 aacatatata tactaactaa ccctgattat ttaaattttc agccaacact tgtcactact
      601 ttctcatggg gcgttcaatg cttctcgaga tacccagatc atatgaaaca gcatgacttt
      661 ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac
      721 gggaactaca agacacgtaa gtttaaacag ttcggtacta actaaccata catatttaaa
      781 ttttcaggtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa
      841 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata caactatatt
      901 tcacacaacg tatacatcac tgccgacaaa caaaagaatg ggatcaaagc ggtaagttta
      961 aacatgattt tactaactaa ctaatctgat ttaaattttc agaacttcaa aattagacac
     1021 aacattgaag atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc
     1081 gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa
     1141 gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt
     1201 acacatggca tggatgaact atacaaatag cattcgtaga attgtaagtt taaactttct
     1261 catactaact aacgcgctct atttaaattt tcagaattcg ctagccggcc atacaagtaa
     1321 tccggatgat cgacgccaac gtcgttgaat tttcaaattt taaatactga atatttgttt
     1381 tttttcctat tatttattta ttctctttgt gttttttttc ttgctttcta aaaaattaat
     1441 tcaatccaaa tctaaacatg agcggttttt tttctctttc cgtctcccaa ttcgtattcc
     1501 gctcctctca tctgaacaca atgtgcaagt ttatttatct tctcgctttc atttcattag
     1561 gacgtggggg gaattggtgg aagggggaaa cacacaaaag gatgatggaa atgaaataag
     1621 gacacacaat atgcaacaac attcaattca gaaatatgga ggaaggttta aaagaaaaca
     1681 taaaaatata tagaggagga aggaaaacta gtaaaaaata agcaaagaaa ttaggcgaac
     1741 gatgagaatt gtcctcgctt gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     1801 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     1861 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     1921 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     1981 gatgcgtaag gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat acgcctattt
     2041 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     2101 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2161 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2221 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2281 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2341 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2401 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2461 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2521 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2581 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2641 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2701 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2761 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2821 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2881 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2941 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3001 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3061 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3121 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3181 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3241 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3301 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3361 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3421 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3481 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3541 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3601 acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa
     3661 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3721 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3781 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3841 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3901 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3961 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     4021 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     4081 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     4141 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     4201 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct gtaagtttaa
     4261 acatgatctt actaactaac tattctcatt taaattttca gagcttaaaa atggctgaaa
     4321 tcactcacaa cgatggatac gctaacaact tggaaatgaa at
//
