LOCUS       sequence-9278-ex        4185 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 4185)
  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 4185)
  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: pGC305
FEATURES             Location/Qualifiers
     source          1..4185
                     /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(1625..1647)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1747..1766
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1844..1862)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1930..2034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2035..2103,2104..2895)
                     /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(2253..2272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3066..3654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3555..3574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3808..3825
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3942..3963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3978..3983,3984..4001,4002..4008)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4016..4032
                     /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     4021..4043
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4040..4056
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4040..4056
                     /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 tactttctgt
      481 tatggtgttc aatgcttctc gagataccca gatcatatga aacggcatga 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 tccacacaat ctgccctttc gaaagatccc
     1021 aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat
     1081 ggcatggacg aactatacaa aagcccacaa gctttggaat tcgctagccg gccatacaag
     1141 taatccggat gatcgacgcc aacgtcgttg aattttcaaa ttttaaatac tgaatatttg
     1201 ttttttttcc tattatttat ttattctctt tgtgtttttt ttcttgcttt ctaaaaaatt
     1261 aattcaatcc aaatctaaac atgagcggtt ttttttctct ttccgtctcc caattcgtat
     1321 tccgctcctc tcatctgaac acaatgtgca agtttattta tcttctcgct ttcatttcat
     1381 taggacgtgg ggggaattgg tggaaggggg aaacacacaa aaggatgatg gaaatgaaat
     1441 aaggacacac aatatgcaac aacattcaat tcagaaatat ggaggaaggt ttaaaagaaa
     1501 acataaaaat atatagagga ggaaggaaaa ctagtaaaaa ataagcaaag aaattaggcg
     1561 aacgatgaga attgtcctcg cttgggccct ttcgtctcgc gcgtttcggt gatgacggtg
     1621 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     1681 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     1741 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     1801 acagatgcgt aaggagaaaa taccgcatca ggcggcctta agggcctcgt gatacgccta
     1861 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     1921 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1981 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2041 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2101 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2161 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2221 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2281 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2341 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2401 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2461 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2521 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2581 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2641 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2701 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2761 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2821 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2881 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2941 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3001 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3181 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     3241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3421 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     3481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3721 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3781 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3841 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3901 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     3961 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     4021 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctgtaagtt
     4081 taaacatgat cttactaact aactattctc atttaaattt tcagagctta aaaatggctg
     4141 aaatcactca caacgatgga tacgctaaca acttggaaat gaaat
//
