LOCUS       sequence-9296-ex        4361 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 4361)
  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 4361)
  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: pGC258
FEATURES             Location/Qualifiers
     source          1..4361
                     /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             201..221
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             243..263
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             285..305
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             327..347
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(390..418)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1166..1186
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1801..1823)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1923..1942
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2020..2038)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2106..2210
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2211..2279,2280..3071)
                     /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(2429..2448)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3242..3830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3731..3750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3984..4001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4118..4139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4154..4159,4160..4177,4178..4184)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4192..4208
                     /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     4197..4219
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4216..4232
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4216..4232
                     /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 ccccgggtat gtttcgaatg atactaacat aacatagaac attttcagga
      181 gctcagaaaa aatgactgct ccaaagaaga agcgtaaggt accggcagaa aaaatgactg
      241 ctccaaagaa gaagcgtaag gtaccggcag aaaaaatgac tgctccaaag aagaagcgta
      301 aggtaccggc agaaaaaatg actgctccaa agaagaagcg taaggtaccg gtagaaaaaa
      361 tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
      421 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
      481 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg gtaagtttaa
      541 acatatatat actaactaac cctgattatt taaattttca gccaacactt gtcactactt
      601 tcggttatgg tcttcaatgc ttcgccagat acccagatca tatgaaacag catgactttt
      661 tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg
      721 ggaactacaa gacacgtaag tttaaacagt tcggtactaa ctaaccatac atatttaaat
      781 tttcaggtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag
      841 gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac aactataact
      901 cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt gtaagtttaa
      961 acatgatttt actaactaac taatctgatt taaattttca gaacttcaaa attagacaca
     1021 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
     1081 atggccctgt ccttttacca gacaaccatt acctgtccta ccaatctgcc ctttcgaaag
     1141 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
     1201 cacatggcat ggatgaacta tacaaatagc attcgtagaa ttgtaagttt aaactttctc
     1261 atactaacta acgcgctcta tttaaatttt cagaattcgc tagccggcca tacaagtaat
     1321 ccggatgatc gacgccaacg tcgttgaatt ttcaaatttt aaatactgaa tatttgtttt
     1381 ttttcctatt atttatttat tctctttgtg ttttttttct tgctttctaa aaaattaatt
     1441 caatccaaat ctaaacatga gcggtttttt ttctctttcc gtctcccaat tcgtattccg
     1501 ctcctctcat ctgaacacaa tgtgcaagtt tatttatctt ctcgctttca tttcattagg
     1561 acgtgggggg aattggtgga agggggaaac acacaaaagg atgatggaaa tgaaataagg
     1621 acacacaata tgcaacaaca ttcaattcag aaatatggag gaaggtttaa aagaaaacat
     1681 aaaaatatat agaggaggaa ggaaaactag taaaaaataa gcaaagaaat taggcgaacg
     1741 atgagaattg tcctcgcttg ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     1801 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1861 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     1921 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     1981 atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg cctcgtgata cgcctatttt
     2041 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     2101 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2161 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2221 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2281 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2341 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2401 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2461 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2521 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2581 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2641 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2701 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2761 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2821 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2881 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2941 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3001 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3061 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3121 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3181 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3241 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3301 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3361 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3421 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3481 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3541 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3601 cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag
     3661 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3721 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3781 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3841 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3901 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3961 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4021 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     4081 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     4141 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     4201 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctg taagtttaaa
     4261 catgatctta ctaactaact attctcattt aaattttcag agcttaaaaa tggctgaaat
     4321 cactcacaac gatggatacg ctaacaactt ggaaatgaaa t
//
