LOCUS       sequence-9300-ex        4140 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 4140)
  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 4140)
  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: pGC302
FEATURES             Location/Qualifiers
     source          1..4140
                     /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"
     CDS             244..264
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             286..306
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             328..348
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             361..1071
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(506..524)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     774..793
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1580..1602)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1702..1721
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1799..1817)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1885..1989
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1990..2058,2059..2850)
                     /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(2208..2227)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3021..3609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3510..3529
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3763..3780
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3897..3918
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3933..3938,3939..3956,3957..3963)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3971..3987
                     /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     3976..3998
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3995..4011
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3995..4011
                     /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 taccggcaga aaaaatgact
      241 gctccaaaga agaagcgtaa ggtaccggca gaaaaaatga ctgctccaaa gaagaagcgt
      301 aaggtaccgg cagaaaaaat gactgctcca aagaagaagc gtaaggtacc ggtagaaaaa
      361 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
      421 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
      481 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
      541 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
      601 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
      661 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
      721 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
      781 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
      841 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
      901 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
      961 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     1021 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta agaattcgct
     1081 agccggccat acaagtaatc cggatgatcg acgccaacgt cgttgaattt tcaaatttta
     1141 aatactgaat atttgttttt tttcctatta tttatttatt ctctttgtgt tttttttctt
     1201 gctttctaaa aaattaattc aatccaaatc taaacatgag cggttttttt tctctttccg
     1261 tctcccaatt cgtattccgc tcctctcatc tgaacacaat gtgcaagttt atttatcttc
     1321 tcgctttcat ttcattagga cgtgggggga attggtggaa gggggaaaca cacaaaagga
     1381 tgatggaaat gaaataagga cacacaatat gcaacaacat tcaattcaga aatatggagg
     1441 aaggtttaaa agaaaacata aaaatatata gaggaggaag gaaaactagt aaaaaataag
     1501 caaagaaatt aggcgaacga tgagaattgt cctcgcttgg gccctttcgt ctcgcgcgtt
     1561 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     1621 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     1681 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     1741 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc
     1801 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     1861 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     1921 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     1981 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2041 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2101 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2161 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2221 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2281 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2341 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2401 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2461 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2521 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2581 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2641 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     2701 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     2761 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     2821 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     2881 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     2941 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3001 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3061 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3121 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3181 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3241 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3301 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3361 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa
     3421 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3481 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3541 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3601 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3661 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3721 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3781 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     3841 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     3901 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     3961 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     4021 gccaagctgt aagtttaaac atgatcttac taactaacta ttctcattta aattttcaga
     4081 gcttaaaaat ggctgaaatc actcacaacg atggatacgc taacaacttg gaaatgaaat
//
