LOCUS       sequence-9304-ex        4978 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 4978)
  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 4978)
  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: pGC244
FEATURES             Location/Qualifiers
     source          1..4978
                     /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    200..323
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(364..669)
                     /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(466..485)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1011..1670)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1585..1604
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1724..1730,1731..1748,1749..1754))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1780..1903)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2418..2440)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2540..2559
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2637..2655)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2723..2827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2828..2896,2897..3688)
                     /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(3046..3065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3859..4447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4348..4367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4601..4618
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4735..4756
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4771..4776,4777..4794,4795..4801)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4809..4825
                     /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     4814..4836
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4833..4849
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4833..4849
                     /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 gctcattacg ccaagctatc aactttgtat agaaaagttg aacgagaaac gtaaaatgat
      241 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
      301 acacaacata tccagtcact atggtcgacc tgcagactgg ctgtgtataa gggagcctga
      361 catttatatt ccccagaaca tcaggttaat ggcgtttttg atgtcatttt cgcggtggct
      421 gagatcagcc acttcttccc cgataacgga gaccggcaca ctggccatat cggtggtcat
      481 catgcgccag ctttcatccc cgatatgcac caccgggtaa agttcacggg ggactttatc
      541 tgacagcaga cgtgcactgg ccagggggat caccatccgt cgcccgggcg tgtcaataat
      601 atcactctgt acatccacaa acagacgata acggctctct cttttatagg tgtaaacctt
      661 aaactgcatt tcaccagccc ctgttctcgt cggcaaaaga gccgttcatt tcaataaacc
      721 gggcgacctc agccatccct tcctgatttt ccgctttcca gcgttcggca cgcagacgac
      781 gggcttcatt ctgcatggtt gtgcttaccg aaccggagat attgacatca tatatgcctt
      841 gagcaactga tagctgtcgc tgtcaactgt cactgtaata cgctgcttca tagcatacct
      901 ctttttgaca tacttcgggt atacatatca gtatatattc ttataccgca aaaatcagcg
      961 cgcaaatacg catactgtta tctggctttt agtaagccgg atcctctaga ttacgccccg
     1021 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
     1081 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     1141 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     1201 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
     1261 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
     1321 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
     1381 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     1441 attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     1501 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     1561 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     1621 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     1681 ttagctcctg aaaatctcga cggatcctaa ctcaaaatcc acacattata cgagccggaa
     1741 gcataaagtg taaagcctgg gggtgcctaa tgcggccgcc atagtgactg gatatgttgt
     1801 gttttacagt attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt
     1861 atatcatttt acgtttctcg ttcaacttta ttatacatag ttgataattc actgggctag
     1921 ccggccatac aagtaatccg gatgatcgac gccaacgtcg ttgaattttc aaattttaaa
     1981 tactgaatat ttgttttttt tcctattatt tatttattct ctttgtgttt tttttcttgc
     2041 tttctaaaaa attaattcaa tccaaatcta aacatgagcg gttttttttc tctttccgtc
     2101 tcccaattcg tattccgctc ctctcatctg aacacaatgt gcaagtttat ttatcttctc
     2161 gctttcattt cattaggacg tggggggaat tggtggaagg gggaaacaca caaaaggatg
     2221 atggaaatga aataaggaca cacaatatgc aacaacattc aattcagaaa tatggaggaa
     2281 ggtttaaaag aaaacataaa aatatataga ggaggaagga aaactagtaa aaaataagca
     2341 aagaaattag gcgaacgatg agaattgtcc tcgcttgggc cctttcgtct cgcgcgtttc
     2401 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     2461 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     2521 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     2581 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcggcc ttaagggcct
     2641 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     2701 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2761 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2821 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2881 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2941 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3001 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3061 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3121 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3181 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3241 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3301 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3361 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3421 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3481 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3541 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3601 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3661 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3721 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3781 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3841 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3901 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3961 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     4021 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     4081 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4141 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4201 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag
     4261 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4321 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4381 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4441 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4501 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4561 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4621 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     4681 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     4741 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     4801 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     4861 caagctgtaa gtttaaacat gatcttacta actaactatt ctcatttaaa ttttcagagc
     4921 ttaaaaatgg ctgaaatcac tcacaacgat ggatacgcta acaacttgga aatgaaat
//
