LOCUS       sequence-9252-ex        7755 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7755)
  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 7755)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pGC158
FEATURES             Location/Qualifiers
     source          1..7755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3302..4573
                     /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"
     primer_bind     complement(5195..5217)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5317..5336
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5414..5432)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5500..5604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5605..5673,5674..6465)
                     /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(5823..5842)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6636..7224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7125..7144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7378..7395
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7512..7533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7548..7553,7554..7571,7572..7578)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7586..7602
                     /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     7591..7613
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7610..7626
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7610..7626
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactagagg atccgcgaca ttgtaggaat catgcgacat
       61 ttctttaaaa ttccccaaaa gctcgtattt ttcaaaattt tggcaatttg ccaaaacttt
      121 gaccggaaat tatgaaaaat ctaaaatttt ttggagtttt ttattatgat attcggttgt
      181 tttggatcat tataagtgca tttagacaaa atccccactg gcgctactcc acctttaaat
      241 tgcagttgaa ttttttttaa attctcattt ttggaaaatt tggaaatgca ccaaaatttt
      301 gactgaaaat tatgaaacat ttgcaatact ttgaaatttg tgtgattcat ttatttagga
      361 cagttattag tttaccttaa atttccctga actactctac tccacttttt ccaccaaatt
      421 attattcacg aacccaaaac tatcttacac atcttatctt acaggttata cattctgcca
      481 cgtcttgtaa ccccctccca cctattctct aatatacaac ctccctttca aacagtccgt
      541 tatgactcta ttgctcctaa tccccctttt gtttccggtt cggcctgaaa caagaataac
      601 aacaggtgac tcgataactg ttaggaaaat aaattttttg gtcccaatat ttcggtatcc
      661 atttcaaaac atcttttaaa attgtcaaga cagacaatta cttctgcttg aaatgaagga
      721 cacatgtccc ctcaattcct cataaaacaa atagaggggt gtcgaaatga taaatgttgc
      781 aagtgaggtg cagatggcaa tgaaatttat atatgtttct ttcattccat tcttgtgggg
      841 caatttcaaa cgtttgataa ctcacagtgg tctagatttg cagaacatct gtgttaggtc
      901 aagcagagcg agaatttgag catttgaact ttagggtcct ggggggggga gggggggggg
      961 gggggggggg tgtctagaca gtcagcggcc cataagggtt catatggttt gacggtgcat
     1021 tgaactattt tttaaggaaa aaattgagat aaaggccatt tttccaagaa agcatctagc
     1081 tttttcaact gacactacaa atttccctat aactttctga aattatcagt gaaaagttgc
     1141 ccaaatttga gactttagct aaaaatagac aattttttcc aaaactttga gcggccataa
     1201 cttttttttg aaaaactttc ggaacgtctc attacaaaat ttggttgctt tgagccattt
     1261 tgaatccgaa aaagtctcaa aaagcaaagt cttaaatttc ggtactccac ctttaaaaaa
     1321 aatagggttt ccgccgaaat ttctgggttc caataatatc tgttaccatt tccttgctct
     1381 gcgacgagca gccaccagac aaaaatctcc taatacccct cttcatgcct ttgttcttcc
     1441 ggtttggcgg tctgaaaaag agcacccctc ccggcggggg gctctcggta tctgttccgc
     1501 aaaaaccgtc atatggaggg ctatcgggag acgggaagag ttggaaatac gggaaatggt
     1561 gatgaatagg tagatcaatc gattccggaa caaataattt gagttgagca cgcgtgctcc
     1621 acatgtgctc agccgcttag gtgtgagtga agatccttgg atttcaaggc catcagccat
     1681 ttcccctgtc cagctagagc agtcctaaac ggaatttttt gcattctttt gcttctttac
     1741 aatttcaaat gttcactcta actaattcaa aaatctttct tttttacgag ctctccttgc
     1801 tcgccgttgt attccattca tacctttaag accacccgag tgtacgcata tttaaccagg
     1861 caccctaaaa acatgaatag tctgtggtgg aggtagacag tgtgttttct aagaatgtct
     1921 ccgtaatttg aaatatattt tagagtcacc tgaaattcat ttttcaagca atatttctta
     1981 ccatatcttt ggggataatt ttaaaagtgg agtgaagtcg tttagggaca cgcttttaag
     2041 ttttaggaaa tggttttttt ttaattcata atttaaaaaa atctataaaa attgttacat
     2101 tttttatgat tttctcaatt ttgttgaatg tttgaatttt ttttttgaga tgtattgttt
     2161 gaattggagc ttttttgtga gttttttttt aattttgcaa ttaaataaaa agtagctaaa
     2221 ttagttttcg aattcccgct caaataaatg tcccagaatt tacatttttg agttttccgg
     2281 ctaaattttt gccgtttttt gaatatgatt tgtaatattt ggtctgaacc atggtttttc
     2341 caattttcag aatctttctc gcaaaaaatc tccgtatccc gcaaaataat tgttgaagcg
     2401 tcaaaaattg gcctaaaagt cgataaatta acattgattt cgcttttttt ttaggaaaaa
     2461 agtctaaaag tattttttaa actatacaaa atcattaaat tttgacaaga aaacgcttaa
     2521 attacaattt tttttatttt ttaaaaattt aaaaaaattt taaaaaaaca ttaaaattca
     2581 gataaattgc acaattttta ttgcacaagt cgttcgtcgt tcgtgttttt ttcgctgtca
     2641 aaatgttggc aaagattgtg aagtcccctg tagtttaaca ctctaagtta ctccaaagac
     2701 ttctacctgc ctacctgcct gcctatctat aacctagtgc ctatcaccta ctcgctgcat
     2761 agtttgatgt acctatataa cagttcataa atgaattttg tcaaaattcc actctcaagt
     2821 attcttacac gtacttattt tgtacaaatc cttgtcagtc gctgcaagaa catacacatc
     2881 gaagaggcgc aatcgaaaca acaggtgttc gcccgttgat cgtctccccg ccccgcctgt
     2941 tgggcgggac gggtgtcggt caccaccacc acatcatatt gttggacaca ctttgcacat
     3001 ccggttcaca cccgattacc gcatcggggt cttgatctgg ggcggctatt ggatcttttg
     3061 tttatgtaga tttttttctc gccgttatga ttttggatta ttttttctct tatcttgcag
     3121 tcgacttgta atctttgcct gattgctagc tagccaaaac tttcacttgt tctttttttt
     3181 cctctaagta ttttcccgaa cttatagtcg accttgagct ttgctagaag ccaacaacac
     3241 ttttcaaatt tgcccgcgga tccccgggtg ccctatagtg agtcgtattg gtaccaaaaa
     3301 catgccacaa tttgatatat tatgtaaaac accacctaag gtgcttgttc gtcagtttgt
     3361 ggaaaggttt gaaagacctt caggtgagaa aatagcatta tgtgctgctg aactaaccta
     3421 tttatgttgg atgattacac ataacggaac agcaatcaag agagccacat tcatgagcta
     3481 taatactatc ataagcaatt cgctgagttt cgatattgtc aataaatcac tccagtttaa
     3541 atacaagacg caaaaagcaa caattctgga agcctcatta aagaaattga ttcctgcttg
     3601 ggaatttaca attattcctt actatggaca aaaacatcaa tctgatatca ctgatattgt
     3661 aagtagtttg caattacagt tcgaatcatc ggaagaagca gataagggaa atagccacag
     3721 taaaaaaatg cttaaagcac ttctaagtga gggtgaaagc atctgggaga tcactgagaa
     3781 aatactaaat tcgtttgagt atacttcgag atttacaaaa acaaaaactt tataccaatt
     3841 cctcttccta gctactttca tcaattgtgg aagattcagc gatattaaga acgttgatcc
     3901 gaaatcattt aaattagtcc aaaataagta tctgggagta ataatccagt gtttagtgac
     3961 agagacaaag acaagcgtta gtaggcacat atacttcttt agcgcaaggg gtaggatcga
     4021 tccacttgta tatttggatg aatttttgag gaattctgaa ccagtcctaa aacgagtaaa
     4081 taggaccggc aattcttcaa gcaataaaca ggaataccaa ttattaaaag ataacttagt
     4141 cagatcgtac aataaagctt tgaagaaaaa tgcgccttat tcaatctttg ctataaaaaa
     4201 tggcccaaaa tctcacattg gaagacattt gatgacctca tttctttcaa tgaagggcct
     4261 aacggagttg actaatgttg tgggaaattg gagcgataag cgtgcttctg ccgtggccag
     4321 gacaacgtat actcatcaga taacagcaat acctgatcac tacttcgcac tagtttctcg
     4381 gtactatgca tatgatccaa tatcaaagga aatgatagca ttgaaggatg agactaatcc
     4441 aattgaggag tggcagcata tagaacagct aaagggtagt gctgaaggaa gcatacgata
     4501 ccccgcatgg aatgggataa tatcacagga ggtactagac tacctttcat cctacataaa
     4561 tagacgcata taagtacgca tttaagcata aacacgcact atgccgttct tctcatgtat
     4621 atatatatac aggcaacacg cagatatagg tgcgacgtga acagtgagct gtatgtgcgg
     4681 ggatccacta gttctagccg gccatacaag taatccggat gatcgacgcc aacgtcgttg
     4741 aattttcaaa ttttaaatac tgaatatttg ttttttttcc tattatttat ttattctctt
     4801 tgtgtttttt ttcttgcttt ctaaaaaatt aattcaatcc aaatctaaac atgagcggtt
     4861 ttttttctct ttccgtctcc caattcgtat tccgctcctc tcatctgaac acaatgtgca
     4921 agtttattta tcttctcgct ttcatttcat taggacgtgg ggggaattgg tggaaggggg
     4981 aaacacacaa aaggatgatg gaaatgaaat aaggacacac aatatgcaac aacattcaat
     5041 tcagaaatat ggaggaaggt ttaaaagaaa acataaaaat atatagagga ggaaggaaaa
     5101 ctagtaaaaa ataagcaaag aaattaggcg aacgatgaga attgtcctcg cttgggccct
     5161 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     5221 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     5281 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     5341 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     5401 ggcggcctta agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5461 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5521 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5581 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5641 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5701 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5761 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5821 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5881 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5941 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6001 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6061 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6121 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6181 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6241 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6301 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6361 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6421 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6481 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6541 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6601 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6661 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6721 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6781 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6841 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6901 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6961 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7021 cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7081 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7141 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7201 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7261 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7321 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7381 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7441 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7501 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7561 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7621 tatgaccatg attacgccaa gctgtaagtt taaacatgat cttactaact aactattctc
     7681 atttaaattt tcagagctta aaaatggctg aaatcactca caacgatgga tacgctaaca
     7741 acttggaaat gaaat
//
