LOCUS       sequence-9306-ex        8833 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 8833)
  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 8833)
  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: pGC260
FEATURES             Location/Qualifiers
     source          1..8833
                     /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"
     CDS             complement(4538..4549)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        6584..6688
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6689..6757,6758..7549)
                     /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(6907..6926)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7720..8308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8209..8228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8462..8479
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8596..8617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8632..8637,8638..8655,8656..8662)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8670..8686
                     /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     8675..8697
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8694..8710
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8694..8710
                     /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 tcttagacaa
     4441 cggataaaat gtttattata atatcaaaaa caatttgtca gttgaccact ttttgatata
     4501 tttgacagaa accgggatga attggctcaa agttagggcg cccttctatt acagggtttc
     4561 tgataacaaa ccggttatta actcccaaca agggatgatt tcaattcatc atgctcaaat
     4621 tgacccaaat taagttacat gacaaattca tcgccctttt caactcttct tggctcatca
     4681 tctgttattc tgttcctatt ctgtctgcac cctatacccc tttgcatact ctctcgtcat
     4741 tcctttcttg gatagtgtct tcttctcccc agctctgcta cttctatgac ttgcgcccgc
     4801 tgcttttccg ctcgttcctc tcctctctga cgtcttcttc tgcttctttt ctccttcctt
     4861 tttcccttct caatcttcct ctttttccca tttctgtcaa tcattcgaag aagaagaaga
     4921 agaagaccct cattcatttc attttttttt ctgtcggtgt gtgctgctcg gttaagagtg
     4981 agctctctat tccacgtctt cttctttttc ttcttcgatt cgaatcaatc actccaccaa
     5041 aacgcattcg tttttgggat tcaccccgcg gttcgcaaca ggtttctttt tcaaattatt
     5101 agcgttataa atagaaaaat ggtggagttt caataaaaat gataatttac aaagtgtatt
     5161 ttgattacat gtactcaaaa aggttgaaat ttttcatacc agttttccgg aaatccatct
     5221 gatatcatta tcgtattttt ctttttaaaa atgttttcaa aaaaaaacaa aatatagctg
     5281 gttatttggc accctctaat taccattttc ctgtcacacc acactctttt ccttcttcca
     5341 cttctttccg ttttcagccg cttccaacca aaccgatatg aaagccgagc aacaacaatc
     5401 gattccaccc ggctcggcga ccttcccgtc gcaggtgaga ctcagaaaac tagagaaacc
     5461 gctcaactaa ctcttgatat ccgatttcat tcttttctct tttctttttg ttgaactttt
     5521 ccatatttcc agatgccacg gccaccacca agcaccgaac aaggaatcac aacggaatcg
     5581 gagcttgcga agaaagctca aatcactccg aacgacgttt tagcacttcc gggaatcact
     5641 caaggtatct tgctcccttt ggatttcagt ttaaaacata aatttaggat tcttatgctc
     5701 cccatctgcg aacatctata acatcgagtt caccaagttc caaatccgtg atctggacac
     5761 tgagcaagtg cttttcgaga tcgccaaacc ggagaacgat caggagaatg atgagtcgcc
     5821 acaggagtcg gcaagatacg tgcgttatag atttgctcca aactttttga aactcaaaac
     5881 ggtcggagca actgtggaat tcaaagtagg agacatccca atccatcatt tccgaatgat
     5941 cgaacgtcac ttcttcaaag atcgccttct gaagtgtttt gactttgaat tcggattctg
     6001 tattccgaat tcacgaaaca actgtgaaca tatctatgag ttccctcaac tctctcaaca
     6061 actcagtgag tcattattct aaaaagtaca attcaaaaga ctaatctctt tttcagtgga
     6121 cgacatgatc aacaatccaa acgagactcg ttctgacagc ttctatttcg tcgataacaa
     6181 actcgtcatg cacaacaaag ccgactactc atatgatgca taaatattta atacaaaaat
     6241 gttctggata attattctgt cgaatagaaa aaaaactcca aatgtgatta aattccaata
     6301 attcctgtct agtttgttcc ttccccttcc ccttctcatg ttcaatgcat tcctaagctt
     6361 ttcagttccc cccttgtttc tatatttttt cgctgtcctg tcacactcgc taaaaacact
     6421 aatcacacgg aaatctgttt tcaataaaaa ctccaacttt aactcatttt caatttcaac
     6481 tgaaagattt tttcattaga gaatgtctag gcatgctcta gagggcccaa aaggcctttt
     6541 ttttttggcg ccgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6661 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6721 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6781 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6841 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6901 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     6961 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     7021 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     7081 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     7141 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     7201 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7261 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7321 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7381 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7441 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7501 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7561 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7861 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     8041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     8101 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     8161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     8221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     8281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     8341 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     8401 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     8461 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     8521 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     8581 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     8641 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     8701 cagctatgac catgattacg ccaagctgta agtttaaaca tgatcttact aactaactat
     8761 tctcatttaa attttcagag cttaaaaatg gctgaaatca ctcacaacga tggatacgct
     8821 aacaacttgg aaa
//
