LOCUS       sequence-9246-ex        6766 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 6766)
  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 6766)
  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: pGC146
FEATURES             Location/Qualifiers
     source          1..6766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             500..1771
                     /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(2471..2482)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4517..4621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4622..4690,4691..5482)
                     /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(4840..4859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5653..6241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6142..6161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6395..6412
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6529..6550
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6565..6570,6571..6588,6589..6595)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6603..6619
                     /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     6608..6630
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6627..6643
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6627..6643
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatcaagag catttgaatc agaatatgga
       61 gaacggagca tgagcatttt cgaagttttt tagatgcact agaacaaagc gtgttggctt
      121 cctctgagcc cgctttcctt atatacccgc attctgcagc cttacagaat gttctagaag
      181 gtcctagatg cattcgtttg aaaatactcc cggtgggtgc aaagagacgc agacggaaaa
      241 tgtatctggg tctctttatt gtgtacacta cttttccatg taccgaatgt gagtcgccct
      301 ccttttgcaa caagcagctc gaatgttcta gaaaaaggtg gaaaatagta taaataccgt
      361 tgaaaataaa taccgaacaa catttgctct aattgtgaaa ttagaaatct tcaaactata
      421 atcatctcac tggatcctat cgattcgcgg ccgctgtaca cccgggtgcc ctatagtgag
      481 tcgtattggt accaaaaaca tgccacaatt tgatatatta tgtaaaacac cacctaaggt
      541 gcttgttcgt cagtttgtgg aaaggtttga aagaccttca ggtgagaaaa tagcattatg
      601 tgctgctgaa ctaacctatt tatgttggat gattacacat aacggaacag caatcaagag
      661 agccacattc atgagctata atactatcat aagcaattcg ctgagtttcg atattgtcaa
      721 taaatcactc cagtttaaat acaagacgca aaaagcaaca attctggaag cctcattaaa
      781 gaaattgatt cctgcttggg aatttacaat tattccttac tatggacaaa aacatcaatc
      841 tgatatcact gatattgtaa gtagtttgca attacagttc gaatcatcgg aagaagcaga
      901 taagggaaat agccacagta aaaaaatgct taaagcactt ctaagtgagg gtgaaagcat
      961 ctgggagatc actgagaaaa tactaaattc gtttgagtat acttcgagat ttacaaaaac
     1021 aaaaacttta taccaattcc tcttcctagc tactttcatc aattgtggaa gattcagcga
     1081 tattaagaac gttgatccga aatcatttaa attagtccaa aataagtatc tgggagtaat
     1141 aatccagtgt ttagtgacag agacaaagac aagcgttagt aggcacatat acttctttag
     1201 cgcaaggggt aggatcgatc cacttgtata tttggatgaa tttttgagga attctgaacc
     1261 agtcctaaaa cgagtaaata ggaccggcaa ttcttcaagc aataaacagg aataccaatt
     1321 attaaaagat aacttagtca gatcgtacaa taaagctttg aagaaaaatg cgccttattc
     1381 aatctttgct ataaaaaatg gcccaaaatc tcacattgga agacatttga tgacctcatt
     1441 tctttcaatg aagggcctaa cggagttgac taatgttgtg ggaaattgga gcgataagcg
     1501 tgcttctgcc gtggccagga caacgtatac tcatcagata acagcaatac ctgatcacta
     1561 cttcgcacta gtttctcggt actatgcata tgatccaata tcaaaggaaa tgatagcatt
     1621 gaaggatgag actaatccaa ttgaggagtg gcagcatata gaacagctaa agggtagtgc
     1681 tgaaggaagc atacgatacc ccgcatggaa tgggataata tcacaggagg tactagacta
     1741 cctttcatcc tacataaata gacgcatata agtacgcatt taagcataaa cacgcactat
     1801 gccgttcttc tcatgtatat atatatacag gcaacacgca gatataggtg cgacgtgaac
     1861 agtgagctgt atgtgcgggg atccactagt tctagccggc catacaagta atccggatga
     1921 tcgacgccaa cgtcgttgaa ttttcaaatt ttaaatactg aatatttgtt ttttttccta
     1981 ttatttattt attctctttg tgtttttttt cttgctttct aaaaaattaa ttcaatccaa
     2041 atctaaacat gagcggtttt ttttctcttt ccgtctccca attcgtattc cgctcctctc
     2101 atctgaacac aatgtgcaag tttatttatc ttctcgcttt catttcatta ggacgtgggg
     2161 ggaattggtg gaagggggaa acacacaaaa ggatgatgga aatgaaataa ggacacacaa
     2221 tatgcaacaa cattcaattc agaaatatgg aggaaggttt aaaagaaaac ataaaaatat
     2281 atagaggagg aaggaaaact agtctagaat cctatgcttg cactttgagc caatttatcc
     2341 aagtccttgt aaaaagtatt cgaaaattgt taacggataa aatgtttatt ataatatcaa
     2401 aaacaatttg tcagttgacc actttttgat atatttgaca gaaaccggga tgaattggct
     2461 caaagttagg gcgcccttct attacagggt ttctgataac aaaccggtta ttaactccca
     2521 acaagggatg atttcaattc atcatgctca aattgaccca aattaagtta catgacaaat
     2581 tcatcgccct tttcaactct tcttggctca tcatctgtta ttctgttcct attctgtctg
     2641 caccctatac ccctttgcat actctctcgt cattcctttc ttggatagtg tcttcttctc
     2701 cccagctctg ctacttctat gacttgcgcc cgctgctttt ccgctcgttc ctctcctctc
     2761 tgacgtcttc ttctgcttct tttctccttc ctttttccct tctcaatctt cctctttttc
     2821 ccatttctgt caatcattcg aagaagaaga agaagaagac cctcattcat ttcatttttt
     2881 tttctgtcgg tgtgtgctgc tcggttaaga gtgagctctc tattccacgt cttcttcttt
     2941 ttcttcttcg attcgaatca atcactccac caaaacgcat tcgtttttgg gattcacccc
     3001 gcggttcgca acaggtttct ttttcaaatt attagcgtta taaatagaaa aatggtggag
     3061 tttcaataaa aatgataatt tacaaagtgt attttgatta catgtactca aaaaggttga
     3121 aatttttcat accagttttc cggaaatcca tctgatatca ttatcgtatt tttcttttta
     3181 aaaatgtttt caaaaaaaaa caaaatatag ctggttattt ggcaccctct aattaccatt
     3241 ttcctgtcac accacactct tttccttctt ccacttcttt ccgttttcag ccgcttccaa
     3301 ccaaaccgat atgaaagccg agcaacaaca atcgattcca cccggctcgg cgaccttccc
     3361 gtcgcaggtg agactcagaa aactagagaa accgctcaac taactcttga tatccgattt
     3421 cattcttttc tcttttcttt ttgttgaact tttccatatt tccagatgcc acggccacca
     3481 ccaagcaccg aacaaggaat cacaacggaa tcggagcttg cgaagaaagc tcaaatcact
     3541 ccgaacgacg ttttagcact tccgggaatc actcaaggta tcttgctccc tttggatttc
     3601 agtttaaaac ataaatttag gattcttatg ctccccatct gcgaacatct ataacatcga
     3661 gttcaccaag ttccaaatcc gtgatctgga cactgagcaa gtgcttttcg agatcgccaa
     3721 accggagaac gatcaggaga atgatgagtc gccacaggag tcggcaagat acgtgcgtta
     3781 tagatttgct ccaaactttt tgaaactcaa aacggtcgga gcaactgtgg aattcaaagt
     3841 aggagacatc ccaatccatc atttccgaat gatcgaacgt cacttcttca aagatcgcct
     3901 tctgaagtgt tttgactttg aattcggatt ctgtattccg aattcacgaa acaactgtga
     3961 acatatctat gagttccctc aactctctca acaactcagt gagtcattat tctaaaaagt
     4021 acaattcaaa agactaatct ctttttcagt ggacgacatg atcaacaatc caaacgagac
     4081 tcgttctgac agcttctatt tcgtcgataa caaactcgtc atgcacaaca aagccgacta
     4141 ctcatatgat gcataaatat ttaatacaaa aatgttctgg ataattattc tgtcgaatag
     4201 aaaaaaaact ccaaatgtga ttaaattcca ataattcctg tctagtttgt tccttcccct
     4261 tccccttctc atgttcaatg cattcctaag cttttcagtt ccccccttgt ttctatattt
     4321 tttcgctgtc ctgtcacact cgctaaaaac actaatcaca cggaaatctg ttttcaataa
     4381 aaactccaac tttaactcat tttcaatttc aactgaaaga ttttttcatt agagaatgtc
     4441 taggcatgct ctagagggcc caaaaggcct tttttttttg gcgccgacgt caggtggcac
     4501 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4561 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4621 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4681 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4741 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4801 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4861 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4921 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     4981 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5041 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5101 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5161 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5221 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5281 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5341 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5401 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5461 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5521 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5581 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5641 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5701 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5761 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     5821 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5881 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5941 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6001 ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac
     6061 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6121 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6181 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6241 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6301 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6361 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6421 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     6481 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     6541 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     6601 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     6661 gtaagtttaa acatgatctt actaactaac tattctcatt taaattttca gagcttaaaa
     6721 atggctgaaa tcactcacaa cgatggatac gctaacaact tggaaa
//
