LOCUS       sequence-9290-ex        3995 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 3995)
  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 3995)
  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: pGC306
FEATURES             Location/Qualifiers
     source          1..3995
                     /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             202..222
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      229..238
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             235..912
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(415..435)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     827..850
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1435..1457)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1557..1576
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1654..1672)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1740..1844
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1845..1913,1914..2705)
                     /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(2063..2082)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2876..3464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3365..3384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3618..3635
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3752..3773
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3788..3793,3794..3811,3812..3818)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3826..3842
                     /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     3831..3853
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3850..3866
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3850..3866
                     /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 ccccggggta tgtttcgaat gatactaaca taacatagaa cattttcagg
      181 agctcagaaa aaatgactgc tccaaagaag aagcgtaagg taccggtcgc caccatggcc
      241 tcctccgaga acgtcatcac cgagttcatg cgcttcaagg tgcgcatgga gggcaccgtg
      301 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggccacaac
      361 accgtgaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
      421 ccccagttcc agtacggctc caaggtgtac gtgaagcacc ccgccgacat ccccgactac
      481 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
      541 ggcgtggcga ccgtgaccca ggactcctcc ctgcaggacg gctgcttcat ctacaaggtg
      601 aagttcatcg gcgtgaactt cccctccgac ggccccgtga tgcagaagaa gaccatgggc
      661 tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagacccac
      721 aaggccctga agctgaagga cggcggccac tacctggtgg agttcaagtc catctacatg
      781 gccaagaagc ccgtgcagct gcccggctac tactacgtgg acgccaagct ggacatcacc
      841 tcccacaacg aggactacac catcgtggag cagtacgagc gcaccgaggg ccgccaccac
      901 ctgttcctgt agcggccgcg actctagaat tcgctagccg gccatacaag taatccggat
      961 gatcgacgcc aacgtcgttg aattttcaaa ttttaaatac tgaatatttg ttttttttcc
     1021 tattatttat ttattctctt tgtgtttttt ttcttgcttt ctaaaaaatt aattcaatcc
     1081 aaatctaaac atgagcggtt ttttttctct ttccgtctcc caattcgtat tccgctcctc
     1141 tcatctgaac acaatgtgca agtttattta tcttctcgct ttcatttcat taggacgtgg
     1201 ggggaattgg tggaaggggg aaacacacaa aaggatgatg gaaatgaaat aaggacacac
     1261 aatatgcaac aacattcaat tcagaaatat ggaggaaggt ttaaaagaaa acataaaaat
     1321 atatagagga ggaaggaaaa ctagtaaaaa ataagcaaag aaattaggcg aacgatgaga
     1381 attgtcctcg cttgggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     1441 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     1501 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     1561 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     1621 aaggagaaaa taccgcatca ggcggcctta agggcctcgt gatacgccta tttttatagg
     1681 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     1741 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1801 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1861 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1921 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1981 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2041 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2101 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2161 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2221 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2281 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2341 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2401 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2461 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2521 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2581 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2641 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2701 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2761 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2821 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2881 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2941 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3001 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3061 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3121 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3181 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3241 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa
     3301 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3361 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3421 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3481 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3541 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3601 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3661 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3721 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     3781 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     3841 aatttcacac aggaaacagc tatgaccatg attacgccaa gctgtaagtt taaacatgat
     3901 cttactaact aactattctc atttaaattt tcagagctta aaaatggctg aaatcactca
     3961 caacgatgga tacgctaaca acttggaaat gaaat
//
