LOCUS       sequence-9323-ex        6555 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 6555)
  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 6555)
  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: pGC450
FEATURES             Location/Qualifiers
     source          1..6555
                     /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    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4664..4759)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4796..4815)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4797..4815)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4820..4836)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4820..4836)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4938..4957)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4949..5758
                     /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(5016..5035)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5905..6493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6394..6413
                     /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 tcgggcccgc gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttggc ggcgaggtcg gcggtacggg cgaagaactt
      721 gcgagacagg cggatgattg aggcttcaca ctgaaaatgg aaagttaagc taagaattaa
      781 gcagatgttt cgaaaaatat tcaaaaatac aatttgaact agacagtaga cctaagattg
      841 aaaagttctg attttcaggt ggtctcgaaa taaacattgg tacttaaaac atgtgtttgc
      901 aaatgtatag ggtgtcacta aagttttgtg tctcagaatg tagttttgtg acttcactgc
      961 cattggtacg agtcatctca ccatagttcc aagagcagtt ggaagctttt gacagacgtt
     1021 cgagtagagt ccgtcaacag cgtgagagtg aagagcctct tggttttgag caaagaagtg
     1081 ctcgacagct ttacagtttt cacatcccaa cataaccttc tctgacatgg ccatttctac
     1141 gagagctgga aattgaaaaa ttacaggatt tcagatgatc gattaactta ccattttcct
     1201 tcttttcgca tgaatgcatc attgtgcaga ttcctcttcc atcaaattga ttgtaagtca
     1261 agaaaagcac agttgacata tacatatgaa caaagtcatt gcatccatct gtccagttgg
     1321 caaccatctt attgcaggca actgtttgga tgtcatcagc tgttccttgc tgaaaatata
     1381 aaccttttaa aactgcttcc aacatacttt tacattttct tttcaataca tgcagtggtt
     1441 attgaaattt gcaacacaat tttctactgc acttactctc ttatcaatga attcctgaag
     1501 aagagcatca gcggaaagtg tgcactcgaa gcaagacata agctcctctc cgttgcttgt
     1561 cttcaccatt cccatagact tccacatctc agatgctggt tgtttcggag cttcacgagc
     1621 agctggcttg gatgtagcag agcagagtcc catgtttgag cacataagtt ccttttcggt
     1681 gaacatatcc ttcatctcgg tcataagatc tggaatgacg gaggagacaa agatgttgca
     1741 tgccttgtgg gcttttccgg cgctcttgca gacaaagcgt tgaagggtag atttgacctg
     1801 aaacttgaga taattttacc gttcttcctg caaaatttaa ctcactgcat aggtagtaaa
     1861 ttcatcccca acgagttttc cgatctgtgc agttgatgcc tgacattcgt cacaaacttc
     1921 ttgtctctat aaaaattaat tgaaaattaa aaattaatct cccaaaatga caaaccataa
     1981 tgttatcatt agctacgatg gcctctgatt tcttcaagta aagcatggca agacgagcaa
     2041 gtggtgagaa ttgagattca ccgcacattt ggagctttga gcatacagcg gaggtgtcag
     2101 cctgaaaatt cagagtactc tctgaaaaaa tgtcgtaaac ctaccaagta tggagcaagt
     2161 ttgtaggcga tgaagtcaag tttgtcaaga gtctttgagc agtcagattg atgagatgta
     2221 cctacgcaga gcattgaaag agaaaccttc agttgagcca tcttcatgcg atccttagaa
     2281 gcatcaatga agttttgaac catctgaaat ttgtttactt tatagtagta acatttagtt
     2341 ctctgaatat aattacttac cgatttgcat gattcacatg gagatggagc agattgtgct
     2401 ccaagagcca aagctgccag cacggcgaag acaaaaatgg atttcattac tgcaatttca
     2461 aatttatttg aaataatttg taagaaatta atgggaaact atgaagaaaa ctgtgcgaaa
     2521 aatgaagcgg aaaactgaaa acacgtgtta ctcggtgctc cttttatcac ctcttatcag
     2581 tgtgctcctc cgacgtgtgc ctctaatctg atttatcagg taatttatgg tgaagcccac
     2641 tatacaacac tagagtatga cccggttttt gattgttgtt aacacgagga aacttcaaaa
     2701 aaatacaaaa attccacgaa attgtttatc acaggtaatt gaaaaagtca tcattgtgct
     2761 cactgaagct tgaaaaaaaa agagcaaaaa ttagagcaat gtggtatgcg acgttggtga
     2821 gagaacgttc aaggaaatat gcagatactg gtttttatgg aaattttcac agcgttattt
     2881 tccggtgctt cataaatttg aaacagtctg agtgacactt tggcactcat tcagtactct
     2941 atgagaacca acatacttcg ttttaccttt ctagaaattt aaaacatttc gataattttt
     3001 ctgcaactgc cgaatggtct agtcgacatc tcatctcatt tttcaactac tataactttt
     3061 tagacttttt ataagaaagc cagtaatgtc gtaagcttta caattttcaa cttgtggata
     3121 aataattgtg aactttttca aaataacaag cgcaataact tcaaatccac ataatgacat
     3181 accacgtttc ctctacttgt tcaatagtct cacactgata ctgagaagtc ttacgtttca
     3241 gaaaggttgg tagagttgcc attaaaaact tcaatgtgct cgaaatgctg ataaattttg
     3301 gaaagttctg tgtgtttttt tacttttaaa cggtgtcgaa tgaaggtttt gtaaaacatc
     3361 gattgttttt gccactaata aggtatgttt tcattgtcca taagttattg atcaactttc
     3421 aatttttctg atagttcagc aatgttgtag ctatttgtca tgaaaatgat ttattcttta
     3481 aacagtttat cagttatcag caagcagaga aatgaagaag tttgatgttt tcaggtctag
     3541 cagttcatct ggaatatttt aattaaaatc gagcattcga atacccaaac tcacagcctg
     3601 actgaatttg ttttggcttg caaggactgc ggtcaaactg acaaatcttg gagcaacaga
     3661 tgacatgatt tgttgacact gaaaattgag atttaagctt tagaacatta aaaaaaacta
     3721 ataaaaactt acacttggtt ggaagaatga taataatcca gagcagaaag tctcagtcag
     3781 tgagttttcg aagaatttct ccatggcact tccaacaagg tttgagctca aatatgtttc
     3841 caattccatc attttcggac atttttcagc ttgaatctct tggaatgtga aatcacgaag
     3901 tggggcaact gaaaaatagt cggtcgatta ttggacacct cattattaca gcaaacctga
     3961 acttgctgga caacacttgt tagctagaca aactccttgt ggagttgcct gttcaactgt
     4021 catctgaata acagttggga gatataaatc caagaaatca atacatgaat cagtcatgtt
     4081 ggctgtgaca atcttgtagc atgcgaaatc acggacatca tcggaaagtt tcttctgaaa
     4141 tcactagagg tgaatcattt aaaaatttgt tttggtgtag gtgattgaaa taaacaaata
     4201 cacagttgga ttgttgtttt tatgttacga aactatttta attgtttttg caataaactt
     4261 actctgttag tgatcatgtc tgtttgcatg tcttcaacaa caaacttgca ttcgaaacag
     4321 gacatggcga gagtctgaaa aatgggagtg tattgcttct attcaaattt tgaaaaataa
     4381 gaaaccacat caaattccct ttttcttgca aaataacaac gaaatctctt tccatctata
     4441 aacttatgag caatatcatc atgttccaat tatttaccga ctttgtctcg acaatgtcag
     4501 tagaagggta acatgtgtga agcaggtggc tgttgggacg gcacacgatt ccttttttag
     4561 atggtaaagg gtcagttaca tctgtcattt tcttttcatt actcaactcc tacttggctc
     4621 acaactcgtc attttttata gcgtcatctt acacaatttt attcaacttt attatacaaa
     4681 gttggcatta taaaaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     4741 caaaataaaa tcattatttg gagctccatg gtagcgttaa cgcggccgcg atatccccta
     4801 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     4861 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     4921 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     4981 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     5041 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     5101 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     5161 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     5221 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     5281 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     5341 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     5401 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     5461 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     5521 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     5581 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     5641 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     5701 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     5761 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     5821 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     5881 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5941 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6001 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     6061 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6121 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6181 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6241 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg
     6301 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6361 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6421 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6481 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6541 ttttgctcac atgtt
//
