LOCUS       sequence-9301-ex        4014 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 4014)
  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 4014)
  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: pGC310
FEATURES             Location/Qualifiers
     source          1..4014
                     /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             235..945
                     /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(380..398)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     648..667
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1454..1476)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1576..1595
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1673..1691)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1759..1863
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1864..1932,1933..2724)
                     /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(2082..2101)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2895..3483
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3384..3403
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3637..3654
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3771..3792
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3807..3812,3813..3830,3831..3837)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3845..3861
                     /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     3850..3872
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3869..3885
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3869..3885
                     /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 taccggtaga aaaaatggtg
      241 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
      301 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
      361 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
      421 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
      481 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
      541 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
      601 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
      661 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
      721 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
      781 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
      841 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
      901 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaagaatt cgctagccgg
      961 ccatacaagt aatccggatg atcgacgcca acgtcgttga attttcaaat tttaaatact
     1021 gaatatttgt tttttttcct attatttatt tattctcttt gtgttttttt tcttgctttc
     1081 taaaaaatta attcaatcca aatctaaaca tgagcggttt tttttctctt tccgtctccc
     1141 aattcgtatt ccgctcctct catctgaaca caatgtgcaa gtttatttat cttctcgctt
     1201 tcatttcatt aggacgtggg gggaattggt ggaaggggga aacacacaaa aggatgatgg
     1261 aaatgaaata aggacacaca atatgcaaca acattcaatt cagaaatatg gaggaaggtt
     1321 taaaagaaaa cataaaaata tatagaggag gaaggaaaac tagtaaaaaa taagcaaaga
     1381 aattaggcga acgatgagaa ttgtcctcgc ttgggccctt tcgtctcgcg cgtttcggtg
     1441 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     1501 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     1561 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     1621 aaataccgca cagatgcgta aggagaaaat accgcatcag gcggccttaa gggcctcgtg
     1681 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     1741 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1801 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1861 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1921 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1981 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2041 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2101 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2161 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2221 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2281 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2341 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2401 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2461 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2521 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2581 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2641 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2701 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     2761 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2821 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2881 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2941 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3001 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3061 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3121 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3181 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3241 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc
     3301 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3361 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3421 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3481 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3541 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3601 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3661 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     3721 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     3781 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     3841 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     3901 ctgtaagttt aaacatgatc ttactaacta actattctca tttaaatttt cagagcttaa
     3961 aaatggctga aatcactcac aacgatggat acgctaacaa cttggaaatg aaat
//
