LOCUS       sequence-9251-ex        4771 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 4771)
  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 4771)
  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: pGC188
FEATURES             Location/Qualifiers
     source          1..4771
                     /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..824
                     /gene="mutant version of attP"
                     /label=attP4
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        978..1080
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1081..1740
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1147..1166)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2082..2387
                     /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     2266..2285
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2744..2975)
                     /gene="mutant version of attP"
                     /label=attP3
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3012..3031)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3013..3031)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3036..3052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3036..3052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3154..3173)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3165..3974
                     /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(3232..3251)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4121..4709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4610..4629
                     /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 aaaagttgaa cgagaaacgt aaaatgatat aaatatcaat
      721 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatg
      781 cagtcactat gaatcaacta cttagatggt attagtgacc tgtagaattc gaggtcgact
      841 aagttggcag catcacccga cgcactttgc gccgaataaa tacctgtgac ggaagatcac
      901 ttcgcagaat aaataaatcc tggtgtccct gttgataccg ggaagccctg ggccaacttt
      961 tggcgaaaat gagacgttga tcggcacgta agaggttcca actttcacca taatgaaata
     1021 agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa
     1081 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
     1141 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
     1201 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
     1261 cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat gaaagacggt
     1321 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
     1381 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
     1441 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
     1501 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
     1561 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
     1621 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtctgtga tggcttccat
     1681 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
     1741 tcgcgtggat ccggcttact aaaagccaga taacagtatg cgtatttgcg cgctgatttt
     1801 tgcggtataa gaatatatac tgatatgtat acccgaagta tgtcaaaaag aggtgtgcta
     1861 tgaagcagcg tattacagtg acagttgaca gcgacagcta tcagttgctc aaggcatata
     1921 tgatgtcaat atctccggtc tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg
     1981 tgccgaacgc tggaaagcgg aaaatcagga agggatggct gaggtcgccc ggtttattga
     2041 aatgaacggc tcttttgctg acgagaacag ggactggtga aatgcagttt aaggtttaca
     2101 cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca
     2161 cgcccgggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct
     2221 cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg
     2281 atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg
     2341 aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc
     2401 ttatacacag ccagtctgca ggtcgataca gtagaaatta cagaaacttt atcacgttta
     2461 gtaagtatag aggctgaaaa tccagatgaa gccgaacgac ttgtaagaga aaagtataag
     2521 agttgtgaaa ttgttcttga tgcagatgat tttcaggact atgacactag cgtatatgaa
     2581 taggtagatg tttttatttt gtcacacaaa aaagaggctc gcacctcttt ttcttatttc
     2641 tttttatgat ttaatacggc attgaggaca atagcgagta ggctggatac gacgattccg
     2701 tttgagaaga acatttggaa ggctgtcggt cgagctcgaa ttctacaggt cactaatacc
     2761 atctaagtag ttgattcata gtgactgcat atgttgtgtt ttacagtatt atgtagtctg
     2821 ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc
     2881 aactttatta tacaaagttg gcattataaa aaagcattgc ttatcaattt gttgcaacga
     2941 acaggtcact atcagtcaaa ataaaatcat tatttggagc tccatggtag cgttaacgcg
     3001 gccgcgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     3061 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
     3121 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     3181 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     3241 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     3301 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     3361 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     3421 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     3481 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     3541 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     3601 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     3661 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     3721 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     3781 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     3841 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     3901 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     3961 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     4021 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     4081 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4141 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4201 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4261 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4321 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4381 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4441 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4501 tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4561 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4621 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4681 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4741 tggccttttg ctggcctttt gctcacatgt t
//
