LOCUS       sequence-468193-        4085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector with FlpTag-GFP, an inverted GFP enabling conditional tagging
            and visualization of endogenous proteins via Flp recombinase 
            activity. Codon phase 0..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4085)
  AUTHORS   Fendl S, Vieira RM, Borst A
  TITLE     Conditional protein tagging methods reveal highly specific 
            subcellular distribution of ion channels in motion-sensing neurons.
  JOURNAL   Elife. 2020 Oct 20;9:e62953. doi: 10.7554/eLife.62953.
  PUBMED    33079061
REFERENCE   2  (bases 1 to 4085)
  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"; journalName: "Elife."; 
            date: "2020-10-20"; volume: "9:e62953. doi"; pages: " 
            10.7554/eLife.62953"
COMMENT     Sequence Label: pUC57-FlpTag-GFP-ph0
FEATURES             Location/Qualifiers
     source          1..4085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..47
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     281..298
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    415..436
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(451..456,457..474,475..481)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    489..505
                     /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     494..516
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     513..529
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     513..529
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    579..648
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     protein_bind    672..705
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(1795..1828)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    1938..2007
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(2043..2060)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2043..2059)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2052..2074)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2268..2287)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2387..2409
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2447..2465)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2533..2637
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2638..3453
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2638..3453
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2711..2730)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3624..4085,1..127)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
       61 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      121 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      181 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      241 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      301 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      361 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      421 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      481 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgctcgagc
      541 caagctcggc gcgccattgg gatgatgtag gtcacggtct cgaagccgcg gtgcgggtgc
      601 cagggcgtgc ccttgggctc cccgggcgcg tactccacct cacccatctg gtccatcatg
      661 atggaagacg ggaagttcct attctctaga aagtatagga acttctcttt gcaccattct
      721 aaagaataac agtgataatt tctgggttaa ggcaagaagt tcctattcta tcagaagtat
      781 aggaacttca gtcgatccaa catggcgact tgtcccatcc ccggcatgtt taaatatact
      841 aattattctt gaactaattt taatcaaccg atttatctct cttccgcagg tgggaggttc
      901 cggtggaagc ggaggtagcg gcggatccgg atccgtgtcc aagggcgagg agctgttcac
      961 cggcgtggtg cccatcctgg tggagctgga tggcgacgtg aacggccaca agttcagcgt
     1021 gcgcggcgag ggcgagggcg acgccaccaa cggcaagctg accctgaagt tcatctgcac
     1081 caccggcaag ctgcccgtgc cctggcccac cctggtgacc accctgacct acggcgtgca
     1141 gtgcttcagc cgctaccccg atcacatgaa gcagcacgat ttcttcaaga gcgccatgcc
     1201 cgagggctac gtgcaggagc gcaccatcag cttcaaggat gacggcacct acaagacccg
     1261 cgccgaggtg aagttcgagg gcgataccct ggtgaaccgc atcgagctga agggcatcga
     1321 tttcaaggag gatggcaaca tcctgggcca caagctggag tacaacttca acagccacaa
     1381 cgtgtacatc accgccgata agcagaagaa cggcatcaag gccaacttca agatccgcca
     1441 caatgtggag gatggctccg tgcagctggc cgatcactac cagcagaaca cccccatcgg
     1501 cgacggccca gtgctgctgc ccgataacca ctacctgagc acccagagcg tgctgtccaa
     1561 ggaccccaac gagaagcgcg atcacatggt gctgctggag ttcgtgaccg ccgccggcat
     1621 caccctgggc atggatgagc tgtacaaggg aggtagcggt ggaagcggag gttccggcga
     1681 ggtaagttat tgaacaatgg catcaaatgc cttcatcatc actacccttt agcccttaag
     1741 accccacaat gaccttaccc actcagagaa agatgcgaca agtcgccaga attcgaagtt
     1801 cctatacttt ctagagaata ggaacttcta agaggtttca tattgctaat agcagctaca
     1861 atccagctac cattctgcga agttcctata cttctgatag aataggaact tcagaagtct
     1921 tccatcatga tggaccagat gggtgaggtg gagtacgcgc ccggggagcc caagggcacg
     1981 ccctggcacc cgcaccgcgg cttcgagacc gtgacctaca tcatcccaat acgcgtcaat
     2041 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     2101 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     2161 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     2221 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     2281 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2341 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     2401 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     2461 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     2521 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2581 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2641 agccatattc aacgggaaac gtcttgctct aggccgcgat taaattccaa catggatgct
     2701 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     2761 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     2821 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     2881 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     2941 cctgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     3001 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     3061 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     3121 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     3181 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     3241 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     3301 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     3361 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     3421 cagtttcatt tgatgctcga tgagtttttc taactgtcag accaagttta ctcatatata
     3481 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3541 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3601 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3661 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3721 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3781 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3841 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3901 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3961 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4021 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4081 ggaac
//
