LOCUS       sequence-9288-ex        4120 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 4120)
  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 4120)
  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: pGC256
FEATURES             Location/Qualifiers
     source          1..4120
                     /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             201..221
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             243..263
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             285..305
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             327..347
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      354..363
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             360..1037
                     /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(540..560)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     952..975
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1560..1582)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1682..1701
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1779..1797)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1865..1969
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1970..2038,2039..2830)
                     /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(2188..2207)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3001..3589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3490..3509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3743..3760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3877..3898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3913..3918,3919..3936,3937..3943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3951..3967
                     /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     3956..3978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3975..3991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3975..3991
                     /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 ccccgggtat gtttcgaatg atactaacat aacatagaac attttcagga
      181 gctcagaaaa aatgactgct ccaaagaaga agcgtaaggt accggcagaa aaaatgactg
      241 ctccaaagaa gaagcgtaag gtaccggcag aaaaaatgac tgctccaaag aagaagcgta
      301 aggtaccggc agaaaaaatg actgctccaa agaagaagcg taaggtaccg gtcgccacca
      361 tggcctcctc cgagaacgtc atcaccgagt tcatgcgctt caaggtgcgc atggagggca
      421 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggcc
      481 acaacaccgt gaagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
      541 tgtcccccca gttccagtac ggctccaagg tgtacgtgaa gcaccccgcc gacatccccg
      601 actacaagaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
      661 acggcggcgt ggcgaccgtg acccaggact cctccctgca ggacggctgc ttcatctaca
      721 aggtgaagtt catcggcgtg aacttcccct ccgacggccc cgtgatgcag aagaagacca
      781 tgggctggga ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga
      841 cccacaaggc cctgaagctg aaggacggcg gccactacct ggtggagttc aagtccatct
      901 acatggccaa gaagcccgtg cagctgcccg gctactacta cgtggacgcc aagctggaca
      961 tcacctccca caacgaggac tacaccatcg tggagcagta cgagcgcacc gagggccgcc
     1021 accacctgtt cctgtagcgg ccgcgactct agaattcgct agccggccat acaagtaatc
     1081 cggatgatcg acgccaacgt cgttgaattt tcaaatttta aatactgaat atttgttttt
     1141 tttcctatta tttatttatt ctctttgtgt tttttttctt gctttctaaa aaattaattc
     1201 aatccaaatc taaacatgag cggttttttt tctctttccg tctcccaatt cgtattccgc
     1261 tcctctcatc tgaacacaat gtgcaagttt atttatcttc tcgctttcat ttcattagga
     1321 cgtgggggga attggtggaa gggggaaaca cacaaaagga tgatggaaat gaaataagga
     1381 cacacaatat gcaacaacat tcaattcaga aatatggagg aaggtttaaa agaaaacata
     1441 aaaatatata gaggaggaag gaaaactagt aaaaaataag caaagaaatt aggcgaacga
     1501 tgagaattgt cctcgcttgg gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     1561 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     1621 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     1681 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     1741 tgcgtaagga gaaaataccg catcaggcgg ccttaagggc ctcgtgatac gcctattttt
     1801 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1861 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1921 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1981 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2041 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2101 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2161 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     2221 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2281 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2341 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2401 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2461 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     2521 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2581 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2641 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2701 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2761 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2821 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2881 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2941 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3001 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3061 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     3181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3361 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
     3421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     3781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     3841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     3901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     3961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctgt aagtttaaac
     4021 atgatcttac taactaacta ttctcattta aattttcaga gcttaaaaat ggctgaaatc
     4081 actcacaacg atggatacgc taacaacttg gaaatgaaat
//
