LOCUS       sequence-9316-ex        6516 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 6516)
  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 6516)
  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: pGC291
FEATURES             Location/Qualifiers
     source          1..6516
                     /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    complement(4641..4740)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4757..4776)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4758..4776)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4781..4797)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4781..4797)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4899..4918)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4910..5719
                     /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(4977..4996)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5866..6454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6355..6374
                     /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 atgccaacgt acaaaaaagc
      661 aggctgcggc gaggtcggcg gtacgggcga agaacttgcg agacaggcgg atgattgagg
      721 cttcacactg aaaatggaaa gttaagctaa gaattaagca gatgtttcga aaaatattca
      781 aaaatacaat ttgaactaga cagtagacct aagattgaaa agttctgatt ttcaggtggt
      841 ctcgaaataa acattggtac ttaaaacatg tgtttgcaaa tgtatagggt gtcactaaag
      901 ttttgtgtct cagaatgtag ttttgtgact tcactgccat tggtacgagt catctcacca
      961 tagttccaag agcagttgga agcttttgac agacgttcga gtagagtccg tcaacagcgt
     1021 gagagtgaag agcctcttgg ttttgagcaa agaagtgctc gacagcttta cagttttcac
     1081 atcccaacat aaccttctct gacatggcca tttctacgag agctggaaat tgaaaaatta
     1141 caggatttca gatgatcgat taacttacca ttttccttct tttcgcatga atgcatcatt
     1201 gtgcagattc ctcttccatc aaattgattg taagtcaaga aaagcacagt tgacatatac
     1261 atatgaacaa agtcattgca tccatctgtc cagttggcaa ccatcttatt gcaggcaact
     1321 gtttggatgt catcagctgt tccttgctga aaatataaac cttttaaaac tgcttccaac
     1381 atacttttac attttctttt caatacatgc agtggttatt gaaatttgca acacaatttt
     1441 ctactgcact tactctctta tcaatgaatt cctgaagaag agcatcagcg gaaagtgtgc
     1501 actcgaagca agacataagc tcctctccgt tgcttgtctt caccattccc atagacttcc
     1561 acatctcaga tgctggttgt ttcggagctt cacgagcagc tggcttggat gtagcagagc
     1621 agagtcccat gtttgagcac ataagttcct tttcggtgaa catatccttc atctcggtca
     1681 taagatctgg aatgacggag gagacaaaga tgttgcatgc cttgtgggct tttccggcgc
     1741 tcttgcagac aaagcgttga agggtagatt tgacctgaaa cttgagataa ttttaccgtt
     1801 cttcctgcaa aatttaactc actgcatagg tagtaaattc atccccaacg agttttccga
     1861 tctgtgcagt tgatgcctga cattcgtcac aaacttcttg tctctataaa aattaattga
     1921 aaattaaaaa ttaatctccc aaaatgacaa accataatgt tatcattagc tacgatggcc
     1981 tctgatttct tcaagtaaag catggcaaga cgagcaagtg gtgagaattg agattcaccg
     2041 cacatttgga gctttgagca tacagcggag gtgtcagcct gaaaattcag agtactctct
     2101 gaaaaaatgt cgtaaaccta ccaagtatgg agcaagtttg taggcgatga agtcaagttt
     2161 gtcaagagtc tttgagcagt cagattgatg agatgtacct acgcagagca ttgaaagaga
     2221 aaccttcagt tgagccatct tcatgcgatc cttagaagca tcaatgaagt tttgaaccat
     2281 ctgaaatttg tttactttat agtagtaaca tttagttctc tgaatataat tacttaccga
     2341 tttgcatgat tcacatggag atggagcaga ttgtgctcca agagccaaag ctgccagcac
     2401 ggcgaagaca aaaatggatt tcattactgc aatttcaaat ttatttgaaa taatttgtaa
     2461 gaaattaatg ggaaactatg aagaaaactg tgcgaaaaat gaagcggaaa actgaaaaca
     2521 cgtgttactc ggtgctcctt ttatcacctc ttatcagtgt gctcctccga cgtgtgcctc
     2581 taatctgatt tatcaggtaa tttatggtga agcccactat acaacactag agtatgaccc
     2641 ggtttttgat tgttgttaac acgaggaaac ttcaaaaaaa tacaaaaatt ccacgaaatt
     2701 gtttatcaca ggtaattgaa aaagtcatca ttgtgctcac tgaagcttga aaaaaaaaga
     2761 gcaaaaatta gagcaatgtg gtatgcgacg ttggtgagag aacgttcaag gaaatatgca
     2821 gatactggtt tttatggaaa ttttcacagc gttattttcc ggtgcttcat aaatttgaaa
     2881 cagtctgagt gacactttgg cactcattca gtactctatg agaaccaaca tacttcgttt
     2941 tacctttcta gaaatttaaa acatttcgat aatttttctg caactgccga atggtctagt
     3001 cgacatctca tctcattttt caactactat aactttttag actttttata agaaagccag
     3061 taatgtcgta agctttacaa ttttcaactt gtggataaat aattgtgaac tttttcaaaa
     3121 taacaagcgc aataacttca aatccacata atgacatacc acgtttcctc tacttgttca
     3181 atagtctcac actgatactg agaagtctta cgtttcagaa aggttggtag agttgccatt
     3241 aaaaacttca atgtgctcga aatgctgata aattttggaa agttctgtgt gtttttttac
     3301 ttttaaacgg tgtcgaatga aggttttgta aaacatcgat tgtttttgcc actaataagg
     3361 tatgttttca ttgtccataa gttattgatc aactttcaat ttttctgata gttcagcaat
     3421 gttgtagcta tttgtcatga aaatgattta ttctttaaac agtttatcag ttatcagcaa
     3481 gcagagaaat gaagaagttt gatgttttca ggtctagcag ttcatctgga atattttaat
     3541 taaaatcgag cattcgaata cccaaactca cagcctgact gaatttgttt tggcttgcaa
     3601 ggactgcggt caaactgaca aatcttggag caacagatga catgatttgt tgacactgaa
     3661 aattgagatt taagctttag aacattaaaa aaaactaata aaaacttaca cttggttgga
     3721 agaatgataa taatccagag cagaaagtct cagtcagtga gttttcgaag aatttctcca
     3781 tggcacttcc aacaaggttt gagctcaaat atgtttccaa ttccatcatt ttcggacatt
     3841 tttcagcttg aatctcttgg aatgtgaaat cacgaagtgg ggcaactgaa aaatagtcgg
     3901 tcgattattg gacacctcat tattacagca aacctgaact tgctggacaa cacttgttag
     3961 ctagacaaac tccttgtgga gttgcctgtt caactgtcat ctgaataaca gttgggagat
     4021 ataaatccaa gaaatcaata catgaatcag tcatgttggc tgtgacaatc ttgtagcatg
     4081 cgaaatcacg gacatcatcg gaaagtttct tctgaaatca ctagaggtga atcatttaaa
     4141 aatttgtttt ggtgtaggtg attgaaataa acaaatacac agttggattg ttgtttttat
     4201 gttacgaaac tattttaatt gtttttgcaa taaacttact ctgttagtga tcatgtctgt
     4261 ttgcatgtct tcaacaacaa acttgcattc gaaacaggac atggcgagag tctgaaaaat
     4321 gggagtgtat tgcttctatt caaattttga aaaataagaa accacatcaa attccctttt
     4381 tcttgcaaaa taacaacgaa atctctttcc atctataaac ttatgagcaa tatcatcatg
     4441 ttccaattat ttaccgactt tgtctcgaca atgtcagtag aagggtaaca tgtgtgaagc
     4501 aggtggctgt tgggacggca cacgattcct tttttagatg gtaaagggtc agttacatct
     4561 gtcattttct tttcattact caactcctac ttggctcaca actcgtcatt ttttatagcg
     4621 tcatcttaca caattttatt acccagcttt cttgtacaaa gttggcatta taagaaagca
     4681 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     4741 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     4801 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa taatatcatc
     4861 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     4921 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     4981 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     5041 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     5101 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     5161 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     5221 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     5281 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     5341 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     5401 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     5461 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     5521 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     5581 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     5641 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     5701 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     5761 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     5821 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5881 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5941 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6001 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6061 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6121 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6181 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6241 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6301 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6361 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6421 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6481 gttcctggcc ttttgctggc cttttgctca catgtt
//
