LOCUS       sequence-9248-ex        8041 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 8041)
  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 8041)
  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: pGC180
FEATURES             Location/Qualifiers
     source          1..8041
                     /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             1820..3091
                     /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(3791..3802)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5957..5978
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6141..6245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6246..6314,6315..7106)
                     /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(6464..6483)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7277..7865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7766..7785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8019..8036
                     /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 acctcgaggg
     1801 ggggcccggt accaaaaaca tgccacaatt tgatatatta tgtaaaacac cacctaaggt
     1861 gcttgttcgt cagtttgtgg aaaggtttga aagaccttca ggtgagaaaa tagcattatg
     1921 tgctgctgaa ctaacctatt tatgttggat gattacacat aacggaacag caatcaagag
     1981 agccacattc atgagctata atactatcat aagcaattcg ctgagtttcg atattgtcaa
     2041 taaatcactc cagtttaaat acaagacgca aaaagcaaca attctggaag cctcattaaa
     2101 gaaattgatt cctgcttggg aatttacaat tattccttac tatggacaaa aacatcaatc
     2161 tgatatcact gatattgtaa gtagtttgca attacagttc gaatcatcgg aagaagcaga
     2221 taagggaaat agccacagta aaaaaatgct taaagcactt ctaagtgagg gtgaaagcat
     2281 ctgggagatc actgagaaaa tactaaattc gtttgagtat acttcgagat ttacaaaaac
     2341 aaaaacttta taccaattcc tcttcctagc tactttcatc aattgtggaa gattcagcga
     2401 tattaagaac gttgatccga aatcatttaa attagtccaa aataagtatc tgggagtaat
     2461 aatccagtgt ttagtgacag agacaaagac aagcgttagt aggcacatat acttctttag
     2521 cgcaaggggt aggatcgatc cacttgtata tttggatgaa tttttgagga attctgaacc
     2581 agtcctaaaa cgagtaaata ggaccggcaa ttcttcaagc aataaacagg aataccaatt
     2641 attaaaagat aacttagtca gatcgtacaa taaagctttg aagaaaaatg cgccttattc
     2701 aatctttgct ataaaaaatg gcccaaaatc tcacattgga agacatttga tgacctcatt
     2761 tctttcaatg aagggcctaa cggagttgac taatgttgtg ggaaattgga gcgataagcg
     2821 tgcttctgcc gtggccagga caacgtatac tcatcagata acagcaatac ctgatcacta
     2881 cttcgcacta gtttctcggt actatgcata tgatccaata tcaaaggaaa tgatagcatt
     2941 gaaggatgag actaatccaa ttgaggagtg gcagcatata gaacagctaa agggtagtgc
     3001 tgaaggaagc atacgatacc ccgcatggaa tgggataata tcacaggagg tactagacta
     3061 cctttcatcc tacataaata gacgcatata agtacgcatt taagcataaa cacgcactat
     3121 gccgttcttc tcatgtatat atatatacag gcaacacgca gatataggtg cgacgtgaac
     3181 agtgagctgt atgtgcgggg atccactagt tctagccggc catacaagta atccggatga
     3241 tcgacgccaa cgtcgttgaa ttttcaaatt ttaaatactg aatatttgtt ttttttccta
     3301 ttatttattt attctctttg tgtttttttt cttgctttct aaaaaattaa ttcaatccaa
     3361 atctaaacat gagcggtttt ttttctcttt ccgtctccca attcgtattc cgctcctctc
     3421 atctgaacac aatgtgcaag tttatttatc ttctcgcttt catttcatta ggacgtgggg
     3481 ggaattggtg gaagggggaa acacacaaaa ggatgatgga aatgaaataa ggacacacaa
     3541 tatgcaacaa cattcaattc agaaatatgg aggaaggttt aaaagaaaac ataaaaatat
     3601 atagaggagg aaggaaaact agtctagaat cctatgcttg cactttgagc caatttatcc
     3661 aagtccttgt aaaaagtatt cgaaaattgt taacggataa aatgtttatt ataatatcaa
     3721 aaacaatttg tcagttgacc actttttgat atatttgaca gaaaccggga tgaattggct
     3781 caaagttagg gcgcccttct attacagggt ttctgataac aaaccggtta ttaactccca
     3841 acaagggatg atttcaattc atcatgctca aattgaccca aattaagtta catgacaaat
     3901 tcatcgccct tttcaactct tcttggctca tcatctgtta ttctgttcct attctgtctg
     3961 caccctatac ccctttgcat actctctcgt cattcctttc ttggatagtg tcttcttctc
     4021 cccagctctg ctacttctat gacttgcgcc cgctgctttt ccgctcgttc ctctcctctc
     4081 tgacgtcttc ttctgcttct tttctccttc ctttttccct tctcaatctt cctctttttc
     4141 ccatttctgt caatcattcg aagaagaaga agaagaagac cctcattcat ttcatttttt
     4201 tttctgtcgg tgtgtgctgc tcggttaaga gtgagctctc tattccacgt cttcttcttt
     4261 ttcttcttcg attcgaatca atcactccac caaaacgcat tcgtttttgg gattcacccc
     4321 gcggttcgca acaggtttct ttttcaaatt attagcgtta taaatagaaa aatggtggag
     4381 tttcaataaa aatgataatt tacaaagtgt attttgatta catgtactca aaaaggttga
     4441 aatttttcat accagttttc cggaaatcca tctgatatca ttatcgtatt tttcttttta
     4501 aaaatgtttt caaaaaaaaa caaaatatag ctggttattt ggcaccctct aattaccatt
     4561 ttcctgtcac accacactct tttccttctt ccacttcttt ccgttttcag ccgcttccaa
     4621 ccaaaccgat atgaaagccg agcaacaaca atcgattcca cccggctcgg cgaccttccc
     4681 gtcgcaggtg agactcagaa aactagagaa accgctcaac taactcttga tatccgattt
     4741 cattcttttc tcttttcttt ttgttgaact tttccatatt tccagatgcc acggccacca
     4801 ccaagcaccg aacaaggaat cacaacggaa tcggagcttg cgaagaaagc tcaaatcact
     4861 ccgaacgacg ttttagcact tccgggaatc actcaaggta tcttgctccc tttggatttc
     4921 agtttaaaac ataaatttag gattcttatg ctccccatct gcgaacatct ataacatcga
     4981 gttcaccaag ttccaaatcc gtgatctgga cactgagcaa gtgcttttcg agatcgccaa
     5041 accggagaac gatcaggaga atgatgagtc gccacaggag tcggcaagat acgtgcgtta
     5101 tagatttgct ccaaactttt tgaaactcaa aacggtcgga gcaactgtgg aattcaaagt
     5161 aggagacatc ccaatccatc atttccgaat gatcgaacgt cacttcttca aagatcgcct
     5221 tctgaagtgt tttgactttg aattcggatt ctgtattccg aattcacgaa acaactgtga
     5281 acatatctat gagttccctc aactctctca acaactcagt gagtcattat tctaaaaagt
     5341 acaattcaaa agactaatct ctttttcagt ggacgacatg atcaacaatc caaacgagac
     5401 tcgttctgac agcttctatt tcgtcgataa caaactcgtc atgcacaaca aagccgacta
     5461 ctcatatgat gcataaatat ttaatacaaa aatgttctgg ataattattc tgtcgaatag
     5521 aaaaaaaact ccaaatgtga ttaaattcca ataattcctg tctagtttgt tccttcccct
     5581 tccccttctc atgttcaatg cattcctaag cttttcagtt ccccccttgt ttctatattt
     5641 tttcgctgtc ctgtcacact cgctaaaaac actaatcaca cggaaatctg ttttcaataa
     5701 aaactccaac tttaactcat tttcaatttc aactgaaaga ttttttcatt agagaatgtc
     5761 taggcatgct ctagagggcc caaaaggggc tttccccgtc aagctctaaa tcgggggctc
     5821 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5881 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5941 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6001 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6061 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     6121 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6181 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6241 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6301 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6361 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6421 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6481 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6541 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6601 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6661 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6721 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6781 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6841 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6901 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6961 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7021 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7081 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7141 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7201 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7261 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7321 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7381 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     7441 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7501 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7561 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7621 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7681 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7741 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7801 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7861 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7921 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7981 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     8041 c
//
