LOCUS       sequence-9298-ex        4236 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4236)
  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 4236)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pGC308
FEATURES             Location/Qualifiers
     source          1..4236
                     /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"
     primer_bind     complement(265..293)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1041..1061
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1676..1698)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1798..1817
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1895..1913)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1981..2085
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2086..2154,2155..2946)
                     /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(2304..2323)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3117..3705
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3606..3625
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3859..3876
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3993..4014
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4029..4034,4035..4052,4053..4059)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4067..4083
                     /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     4072..4094
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4091..4107
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4091..4107
                     /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 aaaaatgagt
      241 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
      301 aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt
      361 acccttaaat ttatttgcac tactggaaaa ctacctgttc catgggtaag tttaaacata
      421 tatatactaa ctaaccctga ttatttaaat tttcagccaa cacttgtcac tactttcggt
      481 tatggtcttc aatgcttcgc cagataccca gatcatatga aacagcatga ctttttcaag
      541 agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac
      601 tacaagacac gtaagtttaa acagttcggt actaactaac catacatatt taaattttca
      661 ggtgctgaag tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt
      721 gattttaaag aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac
      781 aatgtataca tcatggcaga caaacaaaag aatggaatca aagttgtaag tttaaacatg
      841 attttactaa ctaactaatc tgatttaaat tttcagaact tcaaaattag acacaacatt
      901 gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc
      961 cctgtccttt taccagacaa ccattacctg tcctaccaat ctgccctttc gaaagatccc
     1021 aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat
     1081 ggcatggatg aactatacaa atagcattcg tagaattgta agtttaaact ttctcatact
     1141 aactaacgcg ctctatttaa attttcagaa ttcgctagcc ggccatacaa gtaatccgga
     1201 tgatcgacgc caacgtcgtt gaattttcaa attttaaata ctgaatattt gttttttttc
     1261 ctattattta tttattctct ttgtgttttt tttcttgctt tctaaaaaat taattcaatc
     1321 caaatctaaa catgagcggt tttttttctc tttccgtctc ccaattcgta ttccgctcct
     1381 ctcatctgaa cacaatgtgc aagtttattt atcttctcgc tttcatttca ttaggacgtg
     1441 gggggaattg gtggaagggg gaaacacaca aaaggatgat ggaaatgaaa taaggacaca
     1501 caatatgcaa caacattcaa ttcagaaata tggaggaagg tttaaaagaa aacataaaaa
     1561 tatatagagg aggaaggaaa actagtaaaa aataagcaaa gaaattaggc gaacgatgag
     1621 aattgtcctc gcttgggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     1681 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     1741 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     1801 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     1861 taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct atttttatag
     1921 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     1981 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2041 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2101 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2161 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2221 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2281 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2341 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2401 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2461 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2521 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2581 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     2641 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2701 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2761 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2821 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2881 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2941 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3001 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3061 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3121 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3181 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3241 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     3301 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3361 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3421 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3481 accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga
     3541 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3601 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3661 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3721 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3781 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3841 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3901 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3961 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     4021 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     4081 caatttcaca caggaaacag ctatgaccat gattacgcca agctgtaagt ttaaacatga
     4141 tcttactaac taactattct catttaaatt ttcagagctt aaaaatggct gaaatcactc
     4201 acaacgatgg atacgctaac aacttggaaa tgaaat
//
