LOCUS       sequence-468190-        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 2..
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-ph2
FEATURES             Location/Qualifiers
     source          1..4085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      138..726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     627..646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     880..897
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1014..1035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1050..1055,1056..1073,1074..1080)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1088..1104
                     /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     1093..1115
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1112..1128
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1112..1128
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1178..1247
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     protein_bind    1271..1304
                     /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(2394..2427)
                     /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    2537..2606
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(2642..2659)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2642..2658)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2651..2673)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2867..2886)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2986..3008
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3046..3064)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3132..3236
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3237..4052
                     /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             3237..4052
                     /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(3310..3329)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
       61 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
      121 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
      181 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      241 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      301 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      361 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      421 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      481 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      541 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      601 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      661 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      721 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      781 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
      841 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      901 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      961 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     1021 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     1081 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgctcgagcc
     1141 aagctcggcg cgccattggg atgatgtagg tcacggtctc gaagccgcgg tgcgggtgcc
     1201 agggcgtgcc cttgggctcc ccgggcgcgt actccacctc acccatctgg tccatcatga
     1261 tggaagacgg gaagttccta ttctctagaa agtataggaa cttctctttg caccattcta
     1321 aagaataaca gtgataattt ctgggttaag gcaagaagtt cctattctat cagaagtata
     1381 ggaacttcag tcgatccaac atggcgactt gtcccatccc cggcatgttt aaatatacta
     1441 attattcttg aactaatttt aatcaaccga tttatctctc ttccgcaggt cgggaggttc
     1501 cggtggaagc ggaggtagcg gcggatccgg atccgtgtcc aagggcgagg agctgttcac
     1561 cggcgtggtg cccatcctgg tggagctgga tggcgacgtg aacggccaca agttcagcgt
     1621 gcgcggcgag ggcgagggcg acgccaccaa cggcaagctg accctgaagt tcatctgcac
     1681 caccggcaag ctgcccgtgc cctggcccac cctggtgacc accctgacct acggcgtgca
     1741 gtgcttcagc cgctaccccg atcacatgaa gcagcacgat ttcttcaaga gcgccatgcc
     1801 cgagggctac gtgcaggagc gcaccatcag cttcaaggat gacggcacct acaagacccg
     1861 cgccgaggtg aagttcgagg gcgataccct ggtgaaccgc atcgagctga agggcatcga
     1921 tttcaaggag gatggcaaca tcctgggcca caagctggag tacaacttca acagccacaa
     1981 cgtgtacatc accgccgata agcagaagaa cggcatcaag gccaacttca agatccgcca
     2041 caatgtggag gatggctccg tgcagctggc cgatcactac cagcagaaca cccccatcgg
     2101 cgacggccca gtgctgctgc ccgataacca ctacctgagc acccagagcg tgctgtccaa
     2161 ggaccccaac gagaagcgcg atcacatggt gctgctggag ttcgtgaccg ccgccggcat
     2221 caccctgggc atggatgagc tgtacaaggg aggtagcggt ggaagcggag gttccggcag
     2281 gtaagttatt gaacaatggc atcaaatgcc ttcatcatca ctacccttta gcccttaaga
     2341 ccccacaatg accttaccca ctcagagaaa gatgcgacaa gtcgccagaa ttcgaagttc
     2401 ctatactttc tagagaatag gaacttctaa gaggtttcat attgctaata gcagctacaa
     2461 tccagctacc attctgcgaa gttcctatac ttctgataga ataggaactt cagaagtctt
     2521 ccatcatgat ggaccagatg ggtgaggtgg agtacgcgcc cggggagccc aagggcacgc
     2581 cctggcaccc gcaccgcggc ttcgagaccg tgacctacat catcccaata cgcgtcaatt
     2641 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     2701 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     2761 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     2821 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     2881 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     2941 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3001 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3061 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3121 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3181 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3241 gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac atggatgctg
     3301 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     3361 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     3421 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     3481 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     3541 ctgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     3601 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     3661 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     3721 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     3781 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     3841 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     3901 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3961 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     4021 agtttcattt gatgctcgat gagtttttct aactgtcaga ccaagtttac tcatatatac
     4081 tttag
//
