LOCUS       sequence-9315-ex        4304 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 4304)
  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 4304)
  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: pGC235
FEATURES             Location/Qualifiers
     source          1..4304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2429..2528)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2545..2564)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2546..2564)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2569..2585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2569..2585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2687..2706)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2698..3507
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2765..2784)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3654..4242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4143..4162
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctac ttgtgccttg attctcctgt tgaacgtgag tgtttttttt ttaatttgaa
      721 aatgttaagc gtgggccaaa atatttcaaa gaatttacat ccggaaaaaa tttttttgta
      781 aattttctga aaatcacagc ttaaaaaatt caaaacaaaa gttaagttag attcataaaa
      841 tttccatttg aaaagacatc taacagatct aagaatacaa caccttctct caaactccac
      901 ttaatcttga aaaataaatc ttaaaatgtc aaaatccctg caataacaaa atcgaaagaa
      961 ggagagggag aaaatctgga aaaaaagatt attcgcactt ctaagcctta aatatccgtc
     1021 tattttcccc tgaattttgc aactttttta tctgaagctc tggtgctctt ccgattcttt
     1081 tcccatacta cttataggga tgagccaatg aataaatgag caaagttccg aaaggtgtct
     1141 gaacttaatg ggctcacttc caaaaaattt tgaagtcata taacattact aaatttcttg
     1201 gtagaaaaaa cccatttgag ctacacattt ttccatgaca ccttgcttat tcaccagttt
     1261 ggcaaacaaa agaatcgggg ggtctcacac aaaaatatat tttgttgatt aaacgatatg
     1321 atgggggtgg ggaacaaatg ggaggctcca tcaaaaagct cttagttcaa tcaagttctg
     1381 aaagtttaaa agaaaacaaa aatcgctatt tctttcattc catttgtccc cctcctcttt
     1441 tacttttttc ttttccacct tcttcttgtt gtcttaaaaa cgcataactt catctggtcc
     1501 ttcgatgaag atctaagcta taatcgcctt atgcacatca catgtacata tccggtttga
     1561 agatttcaac aacccaacca gattttctcg attttcttgg cctcgaaaac ttaacctaga
     1621 aagttttcaa ataaatatat gatattaata aaaattaact tttcacagtt ttgtattccg
     1681 aaagttttta aaacctaatt ttcttcaaaa aaagtttttc tctaggtcaa ttttcaattc
     1741 taatttcaaa tcgcaacact ctcctcccaa ataagtagtt atttgccgcc gctccgatta
     1801 tgattactgt agtggtgggg tgtttataat tgcttatatt aagtaaactt tccttctttt
     1861 tttgggattg ttacttcttg tttgtcaacg gccagtttac ttcctaaaat aagatgtcaa
     1921 atgttgaaca aggggattat caatagtttg aaatgactac ctttagttgt tatagtttta
     1981 aaattagaac attttaatat gttatttttt aaacagtagg aagctgtcga ctttaaattc
     2041 ttccgaataa aaacggagct caataaaatt aatgacgtgg caaaacagtg ctgacagaag
     2101 agggcaaaaa taaaataaaa aacgttttgg cagctactaa catccccaaa caccccacaa
     2161 atgagctatt ttatgagttt tgttaggctc tccctgcctt tcttcacttt tttcttgaag
     2221 ccatgaaccc cttcatgaca cctaactggg tatcattatc catttgtgcg cgtggccctc
     2281 ccctctttct attacttttc cctaaataaa gcttgagctc ccctcctctc acaaaccctg
     2341 taatcttcaa atcaaagtcg acgggtaatt tgggttagag taaagttaat cgagctttca
     2401 tttttcagaa cgtcagcacc aaccaccgac ccagctttct tgtacaaagt tggcattata
     2461 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc
     2521 attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt
     2581 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaata
     2641 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     2701 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
     2761 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
     2821 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     2881 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     2941 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga
     3001 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     3061 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     3121 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     3181 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     3241 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     3301 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     3361 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     3421 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     3481 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag
     3541 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt
     3601 tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3661 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3721 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3781 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3841 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3901 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3961 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4021 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4081 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4141 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4201 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4261 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
