LOCUS       sequence-9303-ex        4941 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 4941)
  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 4941)
  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: pGC155
FEATURES             Location/Qualifiers
     source          1..4941
                     /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)."
     protein_bind    163..286
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(327..632)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(429..448)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(974..1633)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1548..1567
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1687..1693,1694..1711,1712..1717))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1743..1866)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2381..2403)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2503..2522
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2600..2618)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2686..2790
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2791..2859,2860..3651)
                     /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(3009..3028)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3822..4410
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4311..4330
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4564..4581
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4698..4719
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4734..4739,4740..4757,4758..4764)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4772..4788
                     /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     4777..4799
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4796..4812
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4796..4812
                     /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 ccccgggtac cgagctcatt acgccaagct atcaactttg tatagaaaag
      181 ttgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa
      241 aacagactac ataatactgt aaaacacaac atatccagtc actatggtcg acctgcagac
      301 tggctgtgta taagggagcc tgacatttat attccccaga acatcaggtt aatggcgttt
      361 ttgatgtcat tttcgcggtg gctgagatca gccacttctt ccccgataac ggagaccggc
      421 acactggcca tatcggtggt catcatgcgc cagctttcat ccccgatatg caccaccggg
      481 taaagttcac gggggacttt atctgacagc agacgtgcac tggccagggg gatcaccatc
      541 cgtcgcccgg gcgtgtcaat aatatcactc tgtacatcca caaacagacg ataacggctc
      601 tctcttttat aggtgtaaac cttaaactgc atttcaccag cccctgttct cgtcggcaaa
      661 agagccgttc atttcaataa accgggcgac ctcagccatc ccttcctgat tttccgcttt
      721 ccagcgttcg gcacgcagac gacgggcttc attctgcatg gttgtgctta ccgaaccgga
      781 gatattgaca tcatatatgc cttgagcaac tgatagctgt cgctgtcaac tgtcactgta
      841 atacgctgct tcatagcata cctctttttg acatacttcg ggtatacata tcagtatata
      901 ttcttatacc gcaaaaatca gcgcgcaaat acgcatactg ttatctggct tttagtaagc
      961 cggatcctct agattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
     1021 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
     1081 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
     1141 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
     1201 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
     1261 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
     1321 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
     1381 atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc
     1441 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
     1501 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
     1561 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
     1621 tttttttctc cattttagct tccttagctc ctgaaaatct cgacggatcc taactcaaaa
     1681 tccacacatt atacgagccg gaagcataaa gtgtaaagcc tgggggtgcc taatgcggcc
     1741 gccatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
     1801 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact ttattataca
     1861 tagttgataa ttcactgggc tagccggcca tacaagtaat ccggatgatc gacgccaacg
     1921 tcgttgaatt ttcaaatttt aaatactgaa tatttgtttt ttttcctatt atttatttat
     1981 tctctttgtg ttttttttct tgctttctaa aaaattaatt caatccaaat ctaaacatga
     2041 gcggtttttt ttctctttcc gtctcccaat tcgtattccg ctcctctcat ctgaacacaa
     2101 tgtgcaagtt tatttatctt ctcgctttca tttcattagg acgtgggggg aattggtgga
     2161 agggggaaac acacaaaagg atgatggaaa tgaaataagg acacacaata tgcaacaaca
     2221 ttcaattcag aaatatggag gaaggtttaa aagaaaacat aaaaatatat agaggaggaa
     2281 ggaaaactag taaaaaataa gcaaagaaat taggcgaacg atgagaattg tcctcgcttg
     2341 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     2401 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     2461 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     2521 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     2581 gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     2641 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2701 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2761 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2821 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2881 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2941 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3001 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3061 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3121 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3181 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3241 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3301 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3361 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3421 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3481 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3541 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3601 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3661 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3721 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3781 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3841 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3901 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3961 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4021 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4081 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4141 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4201 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4261 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4321 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4381 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4441 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4501 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4561 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4621 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4681 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     4741 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     4801 aacagctatg accatgatta cgccaagctg taagtttaaa catgatctta ctaactaact
     4861 attctcattt aaattttcag agcttaaaaa tggctgaaat cactcacaac gatggatacg
     4921 ctaacaactt ggaaatgaaa t
//
