LOCUS       sequence-9291-ex        3905 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 3905)
  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 3905)
  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: pGC380
FEATURES             Location/Qualifiers
     source          1..3905
                     /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)."
     regulatory      139..148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             145..822
                     /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(325..345)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     737..760
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1345..1367)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1467..1486
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1564..1582)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1650..1754
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1755..1823,1824..2615)
                     /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(1973..1992)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2786..3374
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3275..3294
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3528..3545
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3662..3683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3698..3703,3704..3721,3722..3728)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3736..3752
                     /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     3741..3763
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3760..3776
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3760..3776
                     /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 ccccggtcgc caccatggcc tcctccgaga acgtcatcac cgagttcatg
      181 cgcttcaagg tgcgcatgga gggcaccgtg aacggccacg agttcgagat cgagggcgag
      241 ggcgagggcc gcccctacga gggccacaac accgtgaagc tgaaggtgac caagggcggc
      301 cccctgccct tcgcctggga catcctgtcc ccccagttcc agtacggctc caaggtgtac
      361 gtgaagcacc ccgccgacat ccccgactac aagaagctgt ccttccccga gggcttcaag
      421 tgggagcgcg tgatgaactt cgaggacggc ggcgtggcga ccgtgaccca ggactcctcc
      481 ctgcaggacg gctgcttcat ctacaaggtg aagttcatcg gcgtgaactt cccctccgac
      541 ggccccgtga tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc
      601 cgcgacggcg tgctgaaggg cgagacccac aaggccctga agctgaagga cggcggccac
      661 tacctggtgg agttcaagtc catctacatg gccaagaagc ccgtgcagct gcccggctac
      721 tactacgtgg acgccaagct ggacatcacc tcccacaacg aggactacac catcgtggag
      781 cagtacgagc gcaccgaggg ccgccaccac ctgttcctgt agcggccgcg actctagaat
      841 tcgctagccg gccatacaag taatccggat gatcgacgcc aacgtcgttg aattttcaaa
      901 ttttaaatac tgaatatttg ttttttttcc tattatttat ttattctctt tgtgtttttt
      961 ttcttgcttt ctaaaaaatt aattcaatcc aaatctaaac atgagcggtt ttttttctct
     1021 ttccgtctcc caattcgtat tccgctcctc tcatctgaac acaatgtgca agtttattta
     1081 tcttctcgct ttcatttcat taggacgtgg ggggaattgg tggaaggggg aaacacacaa
     1141 aaggatgatg gaaatgaaat aaggacacac aatatgcaac aacattcaat tcagaaatat
     1201 ggaggaaggt ttaaaagaaa acataaaaat atatagagga ggaaggaaaa ctagtaaaaa
     1261 ataagcaaag aaattaggcg aacgatgaga attgtcctcg cttgggccct ttcgtctcgc
     1321 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     1381 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     1441 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     1501 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta
     1561 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     1621 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1681 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1741 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1801 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1861 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1921 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1981 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2041 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2101 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2161 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2221 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2281 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     2341 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2401 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2461 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2521 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2581 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2641 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2701 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2761 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2821 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2881 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2941 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3001 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3061 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3121 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt
     3181 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3241 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3301 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3361 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3421 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3481 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3541 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     3601 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     3661 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     3721 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     3781 attacgccaa gctgtaagtt taaacatgat cttactaact aactattctc atttaaattt
     3841 tcagagctta aaaatggctg aaatcactca caacgatgga tacgctaaca acttggaaat
     3901 gaaat
//
