LOCUS       sequence-9305-ex        6722 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 6722)
  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 6722)
  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: pGC246
FEATURES             Location/Qualifiers
     source          1..6722
                     /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)."
     protein_bind    193..317
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(746..1051)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(1491..1615)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    2225..2349
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2681..2986)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3523..3647)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4162..4184)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4284..4303
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4381..4399)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4467..4571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4572..4640,4641..5432)
                     /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(4790..4809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5603..6191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6092..6111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6345..6362
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6479..6500
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6515..6520,6521..6538,6539..6545)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6553..6569
                     /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     6558..6580
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6577..6593
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6577..6593
                     /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 ggatccctcg agacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
      241 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
      301 catatccagt cactatgaat caactactta gatggtatta gtgacctgta gtcgaccgac
      361 agccttccaa atgttcttcg ggtgatgctg ccaacttagt cgaccgacag ccttccaaat
      421 gttcttctca aacggaatcg tcgtatccag cctactcgct attgtcctca atgccgtatt
      481 aaatcataaa aagaaataag aaaaagaggt gcgagcctct tttttgtgtg acaaaataaa
      541 aacatctacc tattcatata cgctagtgtc atagtcctga aaatcatctg catcaagaac
      601 aatttcacaa ctcttatact tttctcttac aagtcgttcg gcttcatctg gattttcagc
      661 ctctatactt actaaacgtg ataaagtttc tgtaatttct actgtatcga cctgcagact
      721 ggctgtgtat aagggagcct gacatttata ttccccagaa catcaggtta atggcgtttt
      781 tgatgtcatt ttcgcggtgg ctgagatcag ccacttcttc cccgataacg gagaccggca
      841 cactggccat atcggtggtc atcatgcgcc agctttcatc cccgatatgc accaccgggt
      901 aaagttcacg ggagacttta tctgacagca gacgtgcact ggccaggggg atcaccatcc
      961 gtcgcccggg cgtgtcaata atatcactct gtacatccac aaacagacga taacggctct
     1021 ctcttttata ggtgtaaacc ttaaactgca tttcaccagt ccctgttctc gtcagcaaaa
     1081 gagccgttca tttcaataaa ccgggcgacc tcagccatcc cttcctgatt ttccgctttc
     1141 cagcgttcgg cacgcagacg acgggcttca ttctgcatgg ttgtgcttac cagaccggag
     1201 atattgacat catatatgcc ttgagcaact gatagctgtc gctgtcaact gtcactgtaa
     1261 tacgctgctt catagcacac ctctttttga catacttcgg gtagtgccga tcaacgtctc
     1321 attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca ggatttattt
     1381 attctgcgaa gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg tcgggtgatg
     1441 ctgccaactt agtcgactac aggtcactaa taccatctaa gtagttgatt catagtgact
     1501 ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
     1561 tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggtctcga
     1621 gaggcctgtt agccaactct agagtactcc tacagtaccc ctatagtaca gctacagtac
     1681 ctcgcactat ccctccatca attaatttta cagtaatcct acagtacccc atacagtatc
     1741 actacaacac ctcctactat ccctccacca actacttcca cagttgtcct acagtacccc
     1801 tacagtatca ctgcagtacc tactactatc tcatcaccaa ctactcctac agtaatccta
     1861 aagtaccatg cactattcct ccatcaacta accttacagt aatcctacag taccgctaca
     1921 gtacctcata ctatccctcc accaattaat cttacagttg tcctacagta ccccatacag
     1981 tatcactaca gtacctccta ctatatcacc accaactact tctacagtac tcctacagta
     2041 cctctatagt gcagctacag tacctcatac tatcccacca acaactaata ttacagtaat
     2101 cctacagtac cccatacagt acctctacac tgcccccgcc atgcaatatg aacatatcaa
     2161 tagtagaaaa aaatccaatt acagaggcct tgatcagcgg ccgcgggaat tcgattgggt
     2221 ctagaccact ttgtacaaga aagctgaacg agaaacgtaa aatgatataa atatcaatat
     2281 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca
     2341 gtcactatga atcaactact tagatggtat tagtgacctg tagtcgacta agttggcagc
     2401 atcacccgac gcactttgcg ccgaataaat acctgtgacg gaagatcact tcgcagaata
     2461 aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg
     2521 agacgttgat cggcacattt cacaactctt atacttttct cttacaagtc gttcggcttc
     2581 atctggattt tcagcctcta tacttactaa acgtgataaa gtttctgtaa tttctactgt
     2641 atcgacctgc agactggctg tgtataaggg agcctgacat ttatattccc cagaacatca
     2701 ggttaatggc gtttttgatg tcattttcgc ggtggctgag atcagccact tcttccccga
     2761 taacggagac cggcacactg gccatatcgg tggtcatcat gcgccagctt tcatccccga
     2821 tatgcaccac cgggtaaagt tcacgggaga ctttatctga cagcagacgt gcactggcca
     2881 gggggatcac catccgtcgc ccgggcgtgt caataatatc actctgtaca tccacaaaca
     2941 gacgataacg gctctctctt ttataggtgt aaaccttaaa ctgcatttca ccagtccctg
     3001 ttctcgtcag caaaagagcc gttcatttca ataaaccggg cgacctcagc catcccttcc
     3061 tgattttccg ctttccagcg ttcggcacgc agacgacggg cttcattctg catggttgtg
     3121 cttaccagac cggagatatt gacatcatat atgccttgag caactgatag ctgtcgctgt
     3181 caactgtcac tgtaatacgc tgcttcatag cacacctctt tttgacatac ttcgggtatg
     3241 ttcttgatgc agatgatttt caggactatg acactagcgt atatgaatag gtagatgttt
     3301 ttattttgtc acacaaaaaa gaggctcgca cctctttttc ttatttcttt ttatgattta
     3361 atacggcatt gaggacaata gcgagtaggc tggatacgac gattccgttt gagaagaaca
     3421 tttggaaggc tgtcggtcga ctaagttggc agcatcaccc gaagaacatt tggaaggctg
     3481 tcggtcgact acaggtcact aataccatct aagtagttga ttcatagtga ctggatatgt
     3541 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
     3601 tttatatcat tttacgtttc tcgttcagct tttttgtaca aacttgtcta gacccaatca
     3661 ctagccggcc atacaagtaa tccggatgat cgacgccaac gtcgttgaat tttcaaattt
     3721 taaatactga atatttgttt tttttcctat tatttattta ttctctttgt gttttttttc
     3781 ttgctttcta aaaaattaat tcaatccaaa tctaaacatg agcggttttt tttctctttc
     3841 cgtctcccaa ttcgtattcc gctcctctca tctgaacaca atgtgcaagt ttatttatct
     3901 tctcgctttc atttcattag gacgtggggg gaattggtgg aagggggaaa cacacaaaag
     3961 gatgatggaa atgaaataag gacacacaat atgcaacaac attcaattca gaaatatgga
     4021 ggaaggttta aaagaaaaca taaaaatata tagaggagga aggaaaacta gtaaaaaata
     4081 agcaaagaaa ttaggcgaac gatgagaatt gtcctcgctt gggccctttc gtctcgcgcg
     4141 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4201 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4261 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     4321 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc ggccttaagg
     4381 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4441 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4501 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4561 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4621 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4681 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4741 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4801 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4861 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4921 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4981 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5041 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5101 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5161 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5221 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5281 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5341 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5401 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5461 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5521 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5581 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5641 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5701 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5761 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5821 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5881 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5941 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag
     6001 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6061 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6121 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6181 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6241 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6301 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6361 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6421 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     6481 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     6541 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     6601 acgccaagct gtaagtttaa acatgatctt actaactaac tattctcatt taaattttca
     6661 gagcttaaaa atggctgaaa tcactcacaa cgatggatac gctaacaact tggaaatgaa
     6721 at
//
