LOCUS       sequence-9250-ex        8073 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 8073)
  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 8073)
  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: pGC267
FEATURES             Location/Qualifiers
     source          1..8073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    60..184
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(209..214,215..232,233..239)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             293..952
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(359..378)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1294..1599
                     /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"
     primer_bind     1478..1497
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1640..1764)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1780..1796)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1781..1797)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             1852..3123
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=FLP
                     /note="FLP is a site-specific recombinase from 
                     Saccharomyces cerevisiae. Recombination occurs at FRT 
                     sequences."
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     CDS             complement(3823..3834)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5989..6010
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6173..6277
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6278..6346,6347..7138)
                     /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(6496..6515)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7309..7897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7798..7817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8051..8068
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aacctggctt atcgaaatta atacgactca ctatagggag accggcagat ctgatatcaa
       61 caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa
      121 attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcat
      181 attggcggcc gcattaggca ccccaggctt tacactttat gcttccggct cgtataatgt
      241 gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta aaatggagaa
      301 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
      361 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
      421 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
      481 tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg gtgagctggt
      541 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
      601 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
      661 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
      721 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
      781 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
      841 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
      901 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aaagatctgg
      961 atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt tttgcggtat
     1021 aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc tatgaagcag
     1081 cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata tatgatgtca
     1141 atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg cgtgccgaac
     1201 gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt gaaatgaacg
     1261 gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta cacctataaa
     1321 agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga cacgcccggg
     1381 cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa
     1441 ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac cgatatggcc
     1501 agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg cgaaaatgac
     1561 atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc ccttatacac
     1621 agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt attatgtagt
     1681 ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt acgtttctcg
     1741 ttcagctttc ttgtacaaag tggttgatat caagcttatc gataccgtcg acctcgagta
     1801 tgtttcgaat gatactaaca taacatagaa cattttcagg gtaccaaaaa catgccacaa
     1861 tttgatatat tatgtaaaac accacctaag gtgcttgttc gtcagtttgt ggaaaggttt
     1921 gaaagacctt caggtgagaa aatagcatta tgtgctgctg aactaaccta tttatgttgg
     1981 atgattacac ataacggaac agcaatcaag agagccacat tcatgagcta taatactatc
     2041 ataagcaatt cgctgagttt cgatattgtc aataaatcac tccagtttaa atacaagacg
     2101 caaaaagcaa caattctgga agcctcatta aagaaattga ttcctgcttg ggaatttaca
     2161 attattcctt actatggaca aaaacatcaa tctgatatca ctgatattgt aagtagtttg
     2221 caattacagt tcgaatcatc ggaagaagca gataagggaa atagccacag taaaaaaatg
     2281 cttaaagcac ttctaagtga gggtgaaagc atctgggaga tcactgagaa aatactaaat
     2341 tcgtttgagt atacttcgag atttacaaaa acaaaaactt tataccaatt cctcttccta
     2401 gctactttca tcaattgtgg aagattcagc gatattaaga acgttgatcc gaaatcattt
     2461 aaattagtcc aaaataagta tctgggagta ataatccagt gtttagtgac agagacaaag
     2521 acaagcgtta gtaggcacat atacttcttt agcgcaaggg gtaggatcga tccacttgta
     2581 tatttggatg aatttttgag gaattctgaa ccagtcctaa aacgagtaaa taggaccggc
     2641 aattcttcaa gcaataaaca ggaataccaa ttattaaaag ataacttagt cagatcgtac
     2701 aataaagctt tgaagaaaaa tgcgccttat tcaatctttg ctataaaaaa tggcccaaaa
     2761 tctcacattg gaagacattt gatgacctca tttctttcaa tgaagggcct aacggagttg
     2821 actaatgttg tgggaaattg gagcgataag cgtgcttctg ccgtggccag gacaacgtat
     2881 actcatcaga taacagcaat acctgatcac tacttcgcac tagtttctcg gtactatgca
     2941 tatgatccaa tatcaaagga aatgatagca ttgaaggatg agactaatcc aattgaggag
     3001 tggcagcata tagaacagct aaagggtagt gctgaaggaa gcatacgata ccccgcatgg
     3061 aatgggataa tatcacagga ggtactagac tacctttcat cctacataaa tagacgcata
     3121 taagtacgca tttaagcata aacacgcact atgccgttct tctcatgtat atatatatac
     3181 aggcaacacg cagatatagg tgcgacgtga acagtgagct gtatgtgcgg ggatccacta
     3241 gttctagccg gccatacaag taatccggat gatcgacgcc aacgtcgttg aattttcaaa
     3301 ttttaaatac tgaatatttg ttttttttcc tattatttat ttattctctt tgtgtttttt
     3361 ttcttgcttt ctaaaaaatt aattcaatcc aaatctaaac atgagcggtt ttttttctct
     3421 ttccgtctcc caattcgtat tccgctcctc tcatctgaac acaatgtgca agtttattta
     3481 tcttctcgct ttcatttcat taggacgtgg ggggaattgg tggaaggggg aaacacacaa
     3541 aaggatgatg gaaatgaaat aaggacacac aatatgcaac aacattcaat tcagaaatat
     3601 ggaggaaggt ttaaaagaaa acataaaaat atatagagga ggaaggaaaa ctagtctaga
     3661 atcctatgct tgcactttga gccaatttat ccaagtcctt gtaaaaagta ttcgaaaatt
     3721 gttaacggat aaaatgttta ttataatatc aaaaacaatt tgtcagttga ccactttttg
     3781 atatatttga cagaaaccgg gatgaattgg ctcaaagtta gggcgccctt ctattacagg
     3841 gtttctgata acaaaccggt tattaactcc caacaaggga tgatttcaat tcatcatgct
     3901 caaattgacc caaattaagt tacatgacaa attcatcgcc cttttcaact cttcttggct
     3961 catcatctgt tattctgttc ctattctgtc tgcaccctat acccctttgc atactctctc
     4021 gtcattcctt tcttggatag tgtcttcttc tccccagctc tgctacttct atgacttgcg
     4081 cccgctgctt ttccgctcgt tcctctcctc tctgacgtct tcttctgctt cttttctcct
     4141 tcctttttcc cttctcaatc ttcctctttt tcccatttct gtcaatcatt cgaagaagaa
     4201 gaagaagaag accctcattc atttcatttt tttttctgtc ggtgtgtgct gctcggttaa
     4261 gagtgagctc tctattccac gtcttcttct ttttcttctt cgattcgaat caatcactcc
     4321 accaaaacgc attcgttttt gggattcacc ccgcggttcg caacaggttt ctttttcaaa
     4381 ttattagcgt tataaataga aaaatggtgg agtttcaata aaaatgataa tttacaaagt
     4441 gtattttgat tacatgtact caaaaaggtt gaaatttttc ataccagttt tccggaaatc
     4501 catctgatat cattatcgta tttttctttt taaaaatgtt ttcaaaaaaa aacaaaatat
     4561 agctggttat ttggcaccct ctaattacca ttttcctgtc acaccacact cttttccttc
     4621 ttccacttct ttccgttttc agccgcttcc aaccaaaccg atatgaaagc cgagcaacaa
     4681 caatcgattc cacccggctc ggcgaccttc ccgtcgcagg tgagactcag aaaactagag
     4741 aaaccgctca actaactctt gatatccgat ttcattcttt tctcttttct ttttgttgaa
     4801 cttttccata tttccagatg ccacggccac caccaagcac cgaacaagga atcacaacgg
     4861 aatcggagct tgcgaagaaa gctcaaatca ctccgaacga cgttttagca cttccgggaa
     4921 tcactcaagg tatcttgctc cctttggatt tcagtttaaa acataaattt aggattctta
     4981 tgctccccat ctgcgaacat ctataacatc gagttcacca agttccaaat ccgtgatctg
     5041 gacactgagc aagtgctttt cgagatcgcc aaaccggaga acgatcagga gaatgatgag
     5101 tcgccacagg agtcggcaag atacgtgcgt tatagatttg ctccaaactt tttgaaactc
     5161 aaaacggtcg gagcaactgt ggaattcaaa gtaggagaca tcccaatcca tcatttccga
     5221 atgatcgaac gtcacttctt caaagatcgc cttctgaagt gttttgactt tgaattcgga
     5281 ttctgtattc cgaattcacg aaacaactgt gaacatatct atgagttccc tcaactctct
     5341 caacaactca gtgagtcatt attctaaaaa gtacaattca aaagactaat ctctttttca
     5401 gtggacgaca tgatcaacaa tccaaacgag actcgttctg acagcttcta tttcgtcgat
     5461 aacaaactcg tcatgcacaa caaagccgac tactcatatg atgcataaat atttaataca
     5521 aaaatgttct ggataattat tctgtcgaat agaaaaaaaa ctccaaatgt gattaaattc
     5581 caataattcc tgtctagttt gttccttccc cttccccttc tcatgttcaa tgcattccta
     5641 agcttttcag ttcccccctt gtttctatat tttttcgctg tcctgtcaca ctcgctaaaa
     5701 acactaatca cacggaaatc tgttttcaat aaaaactcca actttaactc attttcaatt
     5761 tcaactgaaa gattttttca ttagagaatg tctaggcatg ctctagaggg cccaaaaggg
     5821 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5881 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     5941 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     6001 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     6061 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     6121 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     6181 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6241 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6301 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6361 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6421 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6481 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6541 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6601 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6661 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6721 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6781 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6841 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6901 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6961 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     7021 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7081 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7141 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7201 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7261 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7321 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7381 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7441 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7501 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7561 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7621 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7681 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7741 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7801 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7861 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7921 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7981 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     8041 gaccgagcgc agcgagtcag tgagcgagga agc
//
