LOCUS       sequence-9272-ex        4986 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 4986)
  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 4986)
  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: pGC75
FEATURES             Location/Qualifiers
     source          1..4986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(917..964)
                     /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             1003..1023
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1066..1094)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1842..1862
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2426..2448)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2548..2567
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2645..2663)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2731..2835
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2836..2904,2905..3696)
                     /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(3054..3073)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3867..4455
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4356..4375
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4609..4626
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4743..4764
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4779..4784,4785..4802,4803..4809)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4817..4833
                     /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     4822..4844
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4841..4857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4841..4857
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttctgg agattgctcc agcagctctt aacagtgtct acgaccgtct tcgtgcaatt
       61 ctcatggaaa ctgaaagatc ggaaaatgca ctggatcgac gtattcagta tatgattgag
      121 actgcaatgc agattcgaaa ggacaaattt gcggtaaggt agaatatata aatagtttat
      181 tagaaaaaaa taaattagaa taatttaaat tcctactagc caatcaggcg acctttttgc
      241 gcatagttct attattgaaa aatttggaga atttctcata ttctcgctcg gaaatctgga
      301 attcgacgag atcttctggc ttctgtgcag ctgcatcgct ttgtgctccc tttctcgctt
      361 gtcttctgtg tacaccaaga accttgttga gttcatcaac tgaatctgtg actggcttgt
      421 tgctcactgg atgcactaga cgactgattc tcgagaaatc agattgagtt gcgattaggg
      481 tgacctagaa attgggaata atacgaactt ttgaaaatat tcaggaggat taaaaaaatt
      541 attctcgaca atcctacaaa tttacttatt gcaccatgtt gctccaacat ttttcattaa
      601 aagttaatga aaaaatgtag aaaatcggaa attggcaatt ttcagaccat ttttaagcat
      661 tttcaaaaaa aaattgcagc tgaaataaat gtcattttca gataaatcga gcgattttct
      721 gttgtctgac actagttttt agttttaaaa aatgttggaa gaacatggtg caataggtaa
      781 tttcatagaa tttccatgtg ttttttttca attaaccaat tatccaaatc ttccaaactc
      841 acattttgcg gagctgggct atcaagaatc tgctgcagcc cgggggatcc actagttcta
      901 gcttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa
      961 cttcaagatc cccgggtacc gagctcagaa aaaatgacgg cacctaaaaa gaaacgcaaa
     1021 gtaccggtag aaaaaatgag taaaggagaa gaacttttca ctggagttgt cccaattctt
     1081 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt
     1141 gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt
     1201 ccatgggtaa gtttaaacat atatatacta actaaccctg attatttaaa ttttcagcca
     1261 acacttgtca ctactttctg ttatggtgtt caatgcttct cgagataccc agatcatatg
     1321 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata
     1381 tttttcaaag atgacgggaa ctacaagaca cgtaagttta aacagttcgg tactaactaa
     1441 ccatacatat ttaaattttc aggtgctgaa gtcaagtttg aaggtgatac ccttgttaat
     1501 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg
     1561 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc
     1621 aaagttgtaa gtttaaacat gattttacta actaactaat ctgatttaaa ttttcagaac
     1681 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     1741 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     1801 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta
     1861 acagctgctg ggattacaca tggcatggac gaactataca aaagcccaca agctttggaa
     1921 ttcgctagcc ggccatacaa gtaatccgga tgatcgacgc caacgtcgtt gaattttcaa
     1981 attttaaata ctgaatattt gttttttttc ctattattta tttattctct ttgtgttttt
     2041 tttcttgctt tctaaaaaat taattcaatc caaatctaaa catgagcggt tttttttctc
     2101 tttccgtctc ccaattcgta ttccgctcct ctcatctgaa cacaatgtgc aagtttattt
     2161 atcttctcgc tttcatttca ttaggacgtg gggggaattg gtggaagggg gaaacacaca
     2221 aaaggatgat ggaaatgaaa taaggacaca caatatgcaa caacattcaa ttcagaaata
     2281 tggaggaagg tttaaaagaa aacataaaaa tatatagagg aggaaggaaa actagtaaaa
     2341 aataagcaaa gaaattaggc gaacgatgag aattgtcctc gcttgggccc tttcgtctcg
     2401 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     2461 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     2521 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     2581 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcggcctt
     2641 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     2701 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2761 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2821 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2881 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2941 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3001 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3061 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3121 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3181 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3241 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3301 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3361 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3421 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3481 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3541 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3601 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3661 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3721 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3781 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3841 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3901 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3961 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     4021 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4081 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4141 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4201 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat
     4261 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4321 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4381 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4441 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4501 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4561 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     4621 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     4681 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     4741 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     4801 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     4861 gattacgcca agctgtaagt ttaaacatga tcttactaac taactattct catttaaatt
     4921 ttcagagctt aaaaatggct gaaatcactc acaacgatgg atacgctaac aacttggaaa
     4981 tgaaat
//
