LOCUS       sequence-468191-        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 1..
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-ph1
FEATURES             Location/Qualifiers
     source          1..4085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     121..139
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(177..199)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     299..318
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     512..534
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     526..543
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     527..543
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     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    758..791
                     /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(1881..1914)
                     /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(2057..2073)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2057..2073)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2070..2092)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2081..2097
                     /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)."
     promoter        complement(join(2105..2111,2112..2129,2130..2135))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2150..2171
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2288..2305)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2459..3047)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2539..2558)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3218..4033)
                     /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             complement(3218..4033)
                     /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     3941..3960
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(join(4034..4085,1..53))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
       61 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
      121 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
      181 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
      241 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
      301 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
      361 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
      421 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
      481 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
      541 agtgaattga cgcgtattgg gatgatgtag gtcacggtct cgaagccgcg gtgcgggtgc
      601 cagggcgtgc ccttgggctc cccgggcgcg tactccacct cacccatctg gtccatcatg
      661 atggaagact tctgaagttc ctattctatc agaagtatag gaacttcgca gaatggtagc
      721 tggattgtag ctgctattag caatatgaaa cctcttagaa gttcctattc tctagaaagt
      781 ataggaactt cgaattctgg cgacttgtcg catctttctc tgagtgggta aggtcattgt
      841 ggggtcttaa gggctaaagg gtagtgatga tgaaggcatt tgatgccatt gttcaataac
      901 ttacctccgg aacctccgct tccaccgcta cctcccttgt acagctcatc catgcccagg
      961 gtgatgccgg cggcggtcac gaactccagc agcaccatgt gatcgcgctt ctcgttgggg
     1021 tccttggaca gcacgctctg ggtgctcagg tagtggttat cgggcagcag cactgggccg
     1081 tcgccgatgg gggtgttctg ctggtagtga tcggccagct gcacggagcc atcctccaca
     1141 ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct gcttatcggc ggtgatgtac
     1201 acgttgtggc tgttgaagtt gtactccagc ttgtggccca ggatgttgcc atcctccttg
     1261 aaatcgatgc ccttcagctc gatgcggttc accagggtat cgccctcgaa cttcacctcg
     1321 gcgcgggtct tgtaggtgcc gtcatccttg aagctgatgg tgcgctcctg cacgtagccc
     1381 tcgggcatgg cgctcttgaa gaaatcgtgc tgcttcatgt gatcggggta gcggctgaag
     1441 cactgcacgc cgtaggtcag ggtggtcacc agggtgggcc agggcacggg cagcttgccg
     1501 gtggtgcaga tgaacttcag ggtcagcttg ccgttggtgg cgtcgccctc gccctcgccg
     1561 cgcacgctga acttgtggcc gttcacgtcg ccatccagct ccaccaggat gggcaccacg
     1621 ccggtgaaca gctcctcgcc cttggacacg gatccggatc cgccgctacc tccgcttcca
     1681 ccggaacctc cgccacctgc ggaagagaga taaatcggtt gattaaaatt agttcaagaa
     1741 taattagtat atttaaacat gccggggatg ggacaagtcg ccatgttgga tcgactgaag
     1801 ttcctatact tctgatagaa taggaacttc ttgccttaac ccagaaatta tcactgttat
     1861 tctttagaat ggtgcaaaga gaagttccta tactttctag agaataggaa cttcccgtct
     1921 tccatcatga tggaccagat gggtgaggtg gagtacgcgc ccggggagcc caagggcacg
     1981 ccctggcacc cgcaccgcgg cttcgagacc gtgacctaca tcatcccaat ggcgcgccga
     2041 gcttggctcg agcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2101 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2161 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2221 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2281 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2341 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2401 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2461 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2521 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2581 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2641 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2701 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2761 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2821 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2881 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2941 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3001 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3061 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3121 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3181 caatctaaag tatatatgag taaacttggt ctgacagtta gaaaaactca tcgagcatca
     3241 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     3301 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     3361 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     3421 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     3481 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     3541 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     3601 gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     3661 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     3721 ttttcccagg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     3781 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     3841 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     3901 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     3961 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     4021 gaatatggct catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4081 tcatg
//
