LOCUS       sequence-9307-ex        6023 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 6023)
  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 6023)
  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: pGC236
FEATURES             Location/Qualifiers
     source          1..6023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    77..201
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(226..231,232..249,250..256)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             310..969
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(376..395)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1311..1616
                     /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     1495..1514
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1657..1781)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1791..1838)
                     /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             1914..1934
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1956..1976
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1998..2018
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2040..2060
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2103..2131)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2879..2899
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3463..3485)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3585..3604
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3682..3700)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3768..3872
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3873..3941,3942..4733)
                     /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(4091..4110)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4904..5492
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5393..5412
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5646..5663
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5780..5801
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5816..5821,5822..5839,5840..5846)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5854..5870
                     /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     5859..5881
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5878..5894
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5878..5894
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ccgcggtttt ggcgcggatc cactagttct
       61 agcttcaaaa gcatcaacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat
      121 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
      181 acaacatatc cagtcatatt ggcggccgca ttaggcaccc caggctttac actttatgct
      241 tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag
      301 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
      361 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
      421 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
      481 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg
      541 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
      601 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
      661 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
      721 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
      781 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
      841 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
      901 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
      961 ggggcgtaaa gatctggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc
     1021 gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga
     1081 ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca
     1141 aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg
     1201 tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg
     1261 gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa atgcagttta
     1321 aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata
     1381 ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag
     1441 ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga
     1501 tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca
     1561 gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt
     1621 caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt
     1681 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata
     1741 tcattttacg tttctcgttc agctttcttg tacaaagtgg ttgatgctct gaagttccta
     1801 tactttctag agaataggaa cttcggaata ggaacttcaa gatccccggg tatgtttcga
     1861 atgatactaa cataacatag aacattttca ggagctcaga aaaaatgact gctccaaaga
     1921 agaagcgtaa ggtaccggca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg
     1981 cagaaaaaat gactgctcca aagaagaagc gtaaggtacc ggcagaaaaa atgactgctc
     2041 caaagaagaa gcgtaaggta ccggtagaaa aaatgagtaa aggagaagaa cttttcactg
     2101 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     2161 gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta
     2221 ctggaaaact acctgttcca tgggtaagtt taaacatata tatactaact aaccctgatt
     2281 atttaaattt tcagccaaca cttgtcacta ctttctgtta tggtgttcaa tgcttctcga
     2341 gatacccaga tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg
     2401 tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt aagtttaaac
     2461 agttcggtac taactaacca tacatattta aattttcagg tgctgaagtc aagtttgaag
     2521 gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca
     2581 ttcttggaca caaattggaa tacaactata actcacacaa tgtatacatc atggcagaca
     2641 aacaaaagaa tggaatcaaa gttgtaagtt taaacatgat tttactaact aactaatctg
     2701 atttaaattt tcagaacttc aaaattagac acaacattga agatggaagc gttcaactag
     2761 cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc
     2821 attacctgtc cacacaatct gccctttcga aagatcccaa cgaaaagaga gaccacatgg
     2881 tccttcttga gtttgtaaca gctgctggga ttacacatgg catggacgaa ctatacaaaa
     2941 gcccacaagc tttggaattc gctagccggc catacaagta atccggatga tcgacgccaa
     3001 cgtcgttgaa ttttcaaatt ttaaatactg aatatttgtt ttttttccta ttatttattt
     3061 attctctttg tgtttttttt cttgctttct aaaaaattaa ttcaatccaa atctaaacat
     3121 gagcggtttt ttttctcttt ccgtctccca attcgtattc cgctcctctc atctgaacac
     3181 aatgtgcaag tttatttatc ttctcgcttt catttcatta ggacgtgggg ggaattggtg
     3241 gaagggggaa acacacaaaa ggatgatgga aatgaaataa ggacacacaa tatgcaacaa
     3301 cattcaattc agaaatatgg aggaaggttt aaaagaaaac ataaaaatat atagaggagg
     3361 aaggaaaact agtaaaaaat aagcaaagaa attaggcgaa cgatgagaat tgtcctcgct
     3421 tgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3481 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3541 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     3601 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     3661 ccgcatcagg cggccttaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3721 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3781 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3841 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3901 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3961 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4021 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4081 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     4141 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     4201 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     4261 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     4321 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4381 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4441 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4501 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4561 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4621 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4681 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4741 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4801 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4861 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4921 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4981 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5041 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5101 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5161 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5221 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5281 acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5341 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5401 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5461 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5521 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5581 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     5641 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     5701 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5761 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     5821 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     5881 gaaacagcta tgaccatgat tacgccaagc tgtaagttta aacatgatct tactaactaa
     5941 ctattctcat ttaaattttc agagcttaaa aatggctgaa atcactcaca acgatggata
     6001 cgctaacaac ttggaaatga aat
//
