LOCUS       sequence-9299-ex        4146 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 4146)
  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 4146)
  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: pGC382
FEATURES             Location/Qualifiers
     source          1..4146
                     /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)."
     primer_bind     complement(175..203)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     951..971
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1586..1608)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1708..1727
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1805..1823)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1891..1995
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1996..2064,2065..2856)
                     /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(2214..2233)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3027..3615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3516..3535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3769..3786
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3903..3924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3939..3944,3945..3962,3963..3969)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3977..3993
                     /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     3982..4004
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4001..4017
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4001..4017
                     /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 ccccggtaga aaaaatgagt aaaggagaag aacttttcac tggagttgtc
      181 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
      241 ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa
      301 ctacctgttc catgggtaag tttaaacata tatatactaa ctaaccctga ttatttaaat
      361 tttcagccaa cacttgtcac tactttcggt tatggtcttc aatgcttcgc cagataccca
      421 gatcatatga aacagcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
      481 agaactatat ttttcaaaga tgacgggaac tacaagacac gtaagtttaa acagttcggt
      541 actaactaac catacatatt taaattttca ggtgctgaag tcaagtttga aggtgatacc
      601 cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga
      661 cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga caaacaaaag
      721 aatggaatca aagttgtaag tttaaacatg attttactaa ctaactaatc tgatttaaat
      781 tttcagaact tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat
      841 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg
      901 tcctaccaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt
      961 gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa atagcattcg
     1021 tagaattgta agtttaaact ttctcatact aactaacgcg ctctatttaa attttcagaa
     1081 ttcgctagcc ggccatacaa gtaatccgga tgatcgacgc caacgtcgtt gaattttcaa
     1141 attttaaata ctgaatattt gttttttttc ctattattta tttattctct ttgtgttttt
     1201 tttcttgctt tctaaaaaat taattcaatc caaatctaaa catgagcggt tttttttctc
     1261 tttccgtctc ccaattcgta ttccgctcct ctcatctgaa cacaatgtgc aagtttattt
     1321 atcttctcgc tttcatttca ttaggacgtg gggggaattg gtggaagggg gaaacacaca
     1381 aaaggatgat ggaaatgaaa taaggacaca caatatgcaa caacattcaa ttcagaaata
     1441 tggaggaagg tttaaaagaa aacataaaaa tatatagagg aggaaggaaa actagtaaaa
     1501 aataagcaaa gaaattaggc gaacgatgag aattgtcctc gcttgggccc tttcgtctcg
     1561 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     1621 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     1681 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     1741 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcggcctt
     1801 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     1861 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1921 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1981 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2041 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2101 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2161 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2221 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2281 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2341 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2401 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2461 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2521 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2581 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2641 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2701 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2761 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2821 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2881 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2941 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3001 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3061 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3121 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     3181 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3241 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3301 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3361 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat
     3421 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3481 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3541 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3601 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3661 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3721 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3781 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3841 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3901 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     3961 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     4021 gattacgcca agctgtaagt ttaaacatga tcttactaac taactattct catttaaatt
     4081 ttcagagctt aaaaatggct gaaatcactc acaacgatgg atacgctaac aacttggaaa
     4141 tgaaat
//
