LOCUS       sequence-9277-ex        4311 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 4311)
  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 4311)
  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: pGC292
FEATURES             Location/Qualifiers
     source          1..4311
                     /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(1751..1773)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1873..1892
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1970..1988)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2056..2160
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2161..2229,2230..3021)
                     /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(2379..2398)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3192..3780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3681..3700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3934..3951
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4068..4089
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4104..4109,4110..4127,4128..4134)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4142..4158
                     /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     4147..4169
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4166..4182
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4166..4182
                     /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 ttctgttatg gtgttcaatg cttctcgaga tacccagatc atatgaaacg 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 caactataac
      901 tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt tgtaagttta
      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 tggacgaact atacaaaagc ccacaagctt tggaattcgc tagccggcca
     1261 tacaagtaat ccggatgatc gacgccaacg tcgttgaatt ttcaaatttt aaatactgaa
     1321 tatttgtttt ttttcctatt atttatttat tctctttgtg ttttttttct tgctttctaa
     1381 aaaattaatt caatccaaat ctaaacatga gcggtttttt ttctctttcc gtctcccaat
     1441 tcgtattccg ctcctctcat ctgaacacaa tgtgcaagtt tatttatctt ctcgctttca
     1501 tttcattagg acgtgggggg aattggtgga agggggaaac acacaaaagg atgatggaaa
     1561 tgaaataagg acacacaata tgcaacaaca ttcaattcag aaatatggag gaaggtttaa
     1621 aagaaaacat aaaaatatat agaggaggaa ggaaaactag taaaaaataa gcaaagaaat
     1681 taggcgaacg atgagaattg tcctcgcttg ggccctttcg tctcgcgcgt ttcggtgatg
     1741 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     1801 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     1861 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     1921 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg cctcgtgata
     1981 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     2041 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2101 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2161 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     2221 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2281 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     2341 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2401 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     2461 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2521 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2581 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2641 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2701 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2761 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2821 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     2881 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2941 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     3001 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     3061 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3121 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3181 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3241 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3301 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     3361 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3421 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3481 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3541 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg
     3601 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3661 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3721 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3781 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3841 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3901 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3961 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     4021 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     4081 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     4141 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctg
     4201 taagtttaaa catgatctta ctaactaact attctcattt aaattttcag agcttaaaaa
     4261 tggctgaaat cactcacaac gatggatacg ctaacaactt ggaaatgaaa t
//
