LOCUS       sequence-9313-ex        5863 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 5863)
  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 5863)
  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: pGC22
FEATURES             Location/Qualifiers
     source          1..5863
                     /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(3988..4087)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4104..4123)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4105..4123)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4128..4144)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4128..4144)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4246..4265)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4257..5066
                     /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(4324..4343)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5213..5801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5702..5721
                     /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 caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgt tcattgactg tggagcctgg aaatggagga gaaagtaaat ttgatgatta
      721 ctaatacttc gctgtcttta gaggacgcgg agattgtgag acttgaaaca ttttataaat
      781 tttttaattt tactcagaaa taaaatactt aatttctaat ttttttaaag attcaacgct
      841 gatcaaattc tctctcatcg ctggctccaa caatccgccg cttctacgat tctccaaact
      901 ccatccaatc tcattaaccg caaggattcg gcacgtgatg ttcaacaaat gtccgagcac
      961 tttaatctga tgaaccgact tgcggacact cgtctttcgg cgagattcga caataaaatg
     1021 acgatgagcg agtgtggaag tgacttggga actgctacta ttcagttagt ttattatttt
     1081 taaatcttta gttctaattt aaaattcttt tcagtggtga cggatccttc ctatccatgg
     1141 gtggcgaacc ttttggcact ttcccacgaa agaaatctgt tggaattgcc atcgaaaaat
     1201 cacgaagtgg agagttcaca ccgccaatca gtcgtgcttc tcctacaact ccacctccat
     1261 cgatgctcaa tttgagcgaa gatcttactg attcaccggt caaaagaaga tctgccgacg
     1321 attcgggaac cttttcgctc ttctctccag catcatcaaa cggagacgat tcgatctgtt
     1381 ctccaccaat ggtgtttgtc gatatgccat ctattcaact gttcggtact ggagcattgc
     1441 ttacttcagt tcaaatgacg cctcgtcaca ccaccgaaga cgacgcatca ttgaagtcgt
     1501 ttgaggacga gcaggaggtt tgtttcgtca gaatctgtaa taattttgaa ctaaactatt
     1561 ttaatttcag aatgcgaatc ccatccatcg catcgagact caggtcaacg tctaaaccaa
     1621 atcttcgtcg ttgctcctcc ttctcattta acacacaacc tctcaccaaa cctctcgaac
     1681 tctatctaag atcgccactt tttttttctg tttttaaaaa tccccttgta tgtagttttg
     1741 tcattttatt tctcttgcaa tcaaatgcac tcttgttctc atttcacacg aatatttcgt
     1801 tttgtgtgtt cgaaaatgtt ggaaaaaatc atctagattt tagtattagt tttttatttt
     1861 caacattttt gaaaaccacc aatccaccat caattgctct acattgtaaa tagcaagcgt
     1921 tgatctgcac catctcttct tgttttcttt ttcattttta ctctttgact tagaaacgtg
     1981 tacagtacgg ggttgatcac attttcgtgc tggaaatgcg ctgaatttgt gaccaactgc
     2041 gacctaattt gcgtgtacaa atgcccatcc gccccctcaa ctctaaaatg cattgctggt
     2101 gttgttgtta ctacgagatc cccaaattta aacagaatgc tggaaatacg ctgttttccc
     2161 atccactcat tgttgacccg atttgttgtt gatctgaaga gttttttatt ctttgcattg
     2221 acatcaattt ctgttatttt ttttgtattt atcatctttt tatcatcttc tttgcctact
     2281 cccgaatccc ctgtaacatc atcaccacat acttttttct aagatctgac aatcattcac
     2341 attagatttt cagcagtttt caagattttt ctagttgcca ctgctgaaaa tttttcttgc
     2401 aaccacgttg cacccttttt ccacatactc cccaccctct aagacctgta aaacacgtgt
     2461 acaaaatcag agataatgtc gagtttccca gtgactagtt gagctctttt gattctcaga
     2521 tcctgaaatt cctgaaaaag ttttcaattc tgaaagtcga gagctctccg agcactgctc
     2581 cctatcggtg tacatacccg ccttttagtc ggaaaattgt tgatttgtga aaagagtact
     2641 gtaggagatt tttctctcgg cagttctcct tttattacct tccatccatc ccgaggatcg
     2701 tttgcaaagc gtgtttgaca tccagtctcc caactttttc tctgattctc ctccatttgt
     2761 tgtacatttt gcttttagtt tctaacgcct aattccaaca tcggccccca gtttcttttc
     2821 gtttagtaga attgacggct aaactttggt ttaccattca gaagtcacct ttaaaatcat
     2881 ggttttgtct gtcatattag gccctctagg ctcagaatac tcagaataag attaggaaaa
     2941 aaccccaaaa aaatgaagta gtttttgtag tttgtcttca tcacggtttg gaatagccaa
     3001 acccatccca aaccagtctt gcaaaacatc tagcttgact taactcccga gaaccccaca
     3061 gaaagaagcc cgtgtacccc caaccgtccc actcaattgt tgtacgatct cttgtgctga
     3121 aaatgtcgaa aatgttggtg aaagctaact tttttcgatt tttccggttt gtcacaaaaa
     3181 tccttccccg cctatctcgt tgctggagaa tacacacaca cgggcctggt tttggtttcc
     3241 ttcttctccc aaatttccca acaaaatttt tcttaattaa cggaatcccg aaatcacttt
     3301 aatttttata tggagttcac gctctatctc ttctcgatac cattttcccc cacgttcttc
     3361 ctgtcctttc ttcttcataa cttattttct ttgtctccca ctttctctct tgtagtaaat
     3421 taatatccct ttcttctgtt ctgctctgaa attctaatgt tatatttatt ttttatagaa
     3481 atgcttcacg aagttcggaa agaatgataa tggtttcgaa tggagaataa actgtgaata
     3541 atatatttaa atgatttttt acatcacatg aaaagaacat acaatgcata taagagtttg
     3601 atactattga ttttatctgc tacctttcac gaaacggaaa ttagccgttt tttcaaattt
     3661 tatgagtaaa aagttgtgga atcgcaagct acgcctcatg tgtgtacaaa aaaataatgt
     3721 aaaccgatca aaaatataaa attctagtga aaatttgata tttcttatta aatcgaaaac
     3781 ttcgattaaa aacatgtttt ttttgtctcg taaatcctac gtctgcaaac accgatgact
     3841 tagactgcgg actaggcgcc aaattctacg cgcaagttgt ccaagctttt tttctcatca
     3901 tttttttctt tttctcccct tttaaaactt ttaaataact tttgttataa aatctaaagt
     3961 ttttctatat ttcagctact tttggtgacc cagctttctt gtacaaagtt ggcattataa
     4021 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     4081 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     4141 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     4201 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     4261 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     4321 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     4381 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     4441 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     4501 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     4561 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     4621 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     4681 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     4741 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     4801 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     4861 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     4921 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     4981 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     5041 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     5101 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     5161 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5221 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5281 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5341 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5401 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5461 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5521 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5581 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     5641 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5701 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5761 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5821 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
