LOCUS       sequence-9281-ex        4795 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 4795)
  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 4795)
  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: pGC179
FEATURES             Location/Qualifiers
     source          1..4795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(78..125)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             164..184
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             206..226
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             248..268
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             290..310
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             332..1009
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFL"
     CDS             1052..1726
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQA
                     LKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHL
                     FL"
     primer_bind     complement(2235..2257)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2357..2376
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2454..2472)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2540..2644
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2645..2713,2714..3505)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2863..2882)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3676..4264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4165..4184
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4418..4435
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4552..4573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4588..4593,4594..4611,4612..4618)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4626..4642
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4631..4653
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4650..4666
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4650..4666
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ccgcggtttt ggcgcggatc cactagttct
       61 agcttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga
      121 acttcaagat ccccgggtac cgagctcaga aaaaatgact gctccaaaga agaagcgtaa
      181 ggtaccggca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg cagaaaaaat
      241 gactgctcca aagaagaagc gtaaggtacc ggcagaaaaa atgactgctc caaagaagaa
      301 gcgtaaggta ccggtagacg ataaggatcc gatggtggcc tcctccgagg acgtcatcaa
      361 agagttcatg cgcttcaagg tgcgcatgga gggctccgtg aacggccacg agttcgagat
      421 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
      481 caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttcc agtacggctc
      541 caaggcgtac gtgaagcacc ccgccgacat ccccgactac aagaagctgt ccttccccga
      601 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
      661 ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagttccgcg gcaccaactt
      721 cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg
      781 cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga
      841 cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcagct
      901 gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac
      961 catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgg ggcatggcac
     1021 cggcagcacc ggcagcggca gctccggcac cgcctcctcc gaggacgtca tcaaagagtt
     1081 catgcgcttc aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     1141 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     1201 cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg gctccaaggc
     1261 gtacgtgaag caccccgccg acatccccga ctacaagaag ctgtccttcc ccgagggctt
     1321 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     1381 ctccctgcag gacggcacgc tgatctacaa ggtgaagttc cgcggcacca acttcccccc
     1441 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta
     1501 cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg
     1561 ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc agctgcccgg
     1621 ctactactac gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt
     1681 ggaacagtac gagcgctccg agggccgcca ccacctgttc ctgtaggaat tcgctagccg
     1741 gccatacaag taatccggat gatcgacgcc aacgtcgttg aattttcaaa ttttaaatac
     1801 tgaatatttg ttttttttcc tattatttat ttattctctt tgtgtttttt ttcttgcttt
     1861 ctaaaaaatt aattcaatcc aaatctaaac atgagcggtt ttttttctct ttccgtctcc
     1921 caattcgtat tccgctcctc tcatctgaac acaatgtgca agtttattta tcttctcgct
     1981 ttcatttcat taggacgtgg ggggaattgg tggaaggggg aaacacacaa aaggatgatg
     2041 gaaatgaaat aaggacacac aatatgcaac aacattcaat tcagaaatat ggaggaaggt
     2101 ttaaaagaaa acataaaaat atatagagga ggaaggaaaa ctagtaaaaa ataagcaaag
     2161 aaattaggcg aacgatgaga attgtcctcg cttgggccct ttcgtctcgc gcgtttcggt
     2221 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     2281 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     2341 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     2401 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta agggcctcgt
     2461 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     2521 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2581 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2641 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2701 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2761 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2821 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2881 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2941 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3001 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3061 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3121 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3181 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3241 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3301 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3361 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3421 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3481 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3541 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3601 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3661 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3721 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3781 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3841 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3901 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3961 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4021 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc
     4081 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4141 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4201 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4261 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4321 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4381 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4441 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     4501 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     4561 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     4621 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     4681 gctgtaagtt taaacatgat cttactaact aactattctc atttaaattt tcagagctta
     4741 aaaatggctg aaatcactca caacgatgga tacgctaaca acttggaaat gaaat
//
