LOCUS       sequence-137517-        9639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TagRFP-T_PH pCFJ150 - contains C. elegans codon optimized TagRFP-T.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9639)
  AUTHORS   Heppert JK, Dickinson DJ, Pani AM, Higgins CD, Steward A, Ahringer 
            J, Kuhn JR, Goldstein B
  TITLE     Comparative assessment of fluorescent proteins for in vivo imaging 
            in an animal model system.
  JOURNAL   Mol Biol Cell. 2016 Nov 7;27(22):3385-3394. doi: 
            10.1091/mbc.E16-01-0063. Epub 2016 Jul 6.
  PUBMED    27385332
REFERENCE   2  (bases 1 to 9639)
  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: 
            "10.1091/mbc.E16-01-0063"; journalName: "Mol Biol Cell"; date: 
            "2016-11-7- 7"; volume: "27"; issue: "22"; pages: "3385-3394"
COMMENT     Sequence Label: CH00X
FEATURES             Location/Qualifiers
     source          1..9639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..21
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             1962..1985
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2001..2024
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      2036..2359
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     primer_bind     complement(3938..3957)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4057..4079
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4117..4135)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4203..4307
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4308..4376,4377..5168)
                     /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(4526..4545)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5339..5927
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5828..5847
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7563..7574)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 caactttgta tagaaaagtt gaaatatcag tttttaaaaa attaaaccat aaaacaaata
       61 atataaccca atttttacat caaaccacaa gaaaaaaata catttgggcc cacggataaa
      121 gaaattaaaa aaatacattt tttaaaggcg caccgaatta aaattcattt gggtcttacc
      181 gcgtataccg tactccgttt gtttgatcat ttttgtcagc gctggcggtt gttttttcat
      241 ttcatttctg cttcaaagac gttttctcga ataatttttc gtttattctc ttttttaaaa
      301 ttaatttcta gccgtaaatg ttataaattc acccatttaa cgcaaatttc atggtaatct
      361 catggaaaaa tgcagtttct ttgttaaaga aagcttaaat agcaaaaatt ccccgacttt
      421 ccccaaaatc ctgctcgatt ttccgttttc tcattgtatt ctctcttaat taattttatc
      481 gataatcaat tgaatgtttc agacagagaa tggtctccaa gggagaggag ctcatcaagg
      541 agaacatgca catgaagctc tacatggagg gaaccgtcaa caaccaccac ttcaagtgca
      601 cctccgaggg agagggaaag ccatacgagg gaacccaaac catgcgtatc aaggtaagtt
      661 taaacatata tatactaact aaccctgatt atttaaattt tcaggtcgtc gagggaggac
      721 cactcccatt cgccttcgac atcctcgcca cctccttcat gtacggatcc cgtaccttca
      781 tcaaccacac ccaaggaatc ccagacttct tcaagcaatc cttcccagag ggattcacct
      841 gggagcgtgt caccacctac gaggacggag gagtcctcac cgccacccaa gacacctccc
      901 tccaagacgg atgcctcatc tacaacgtca aggtaagttt aaacagttcg gtactaacta
      961 accatacata tttaaatttt cagatccgtg gagtcaactt cccatccaac ggaccagtca
     1021 tgcaaaagaa gaccctcgga tgggaggcca acaccgagat gctctaccca gccgacggag
     1081 gactcgaggg acgtaccgac atggccctca agctcgtcgg aggaggacac ctcatctgca
     1141 acttcaagac cacctaccgt tccaaggtaa gtttaaacat gattttacta actaactaat
     1201 ctgatttaaa ttttcagaag ccagccaaga acctcaagat gccaggagtc tactacgtcg
     1261 accaccgtct cgagcgtatc aaggaggccg acaaggagac ctacgtcgag caacacgagg
     1321 tcgccgtcgc ccgttactgc gacctcccat ccaagctcgg acacaagctc aacggaatgg
     1381 acgagctcta caaggccggc cacggactcc aagacgaccc agacctccaa gccctcctca
     1441 agggatccca actcctcaag gtcaagtcct cctcctggcg tcgtgagcgt ttctacaagc
     1501 tccaagagga ctgcaaggta agtttaaaca tatatatact aactaaccct gattatttaa
     1561 attttcagac catctggcaa gagtcccgta aggtaagttt aaacagttcg gtactaacta
     1621 accatacata tttaaatttt caggtcatgc gttccccaga gtcccaactc ttctccatcg
     1681 aggacatcca agaggtccgt atgggacacc gtaccgaggg actcgagaag ttcgcccgtg
     1741 acatcccaga ggaccgttgc ttctccatcg tcttcaaggt aagtttaaac atgattttac
     1801 taactaacta atctgattta aattttcagg accaacgtaa caccctcgac ctcatcgccc
     1861 catccccagc cgacgcccaa cactgggtcc aaggactccg taagatcatc caccactccg
     1921 gatccatgga ccaacgtcaa aagtcaggag ctagcggagc cgattacaag gatgacgatg
     1981 acaagagagg atctggatct gactacaagg acgatgacga taagcgttaa gataaatgca
     2041 aaatcctttc aagcattccc ttcttctcta tcactcttct ttctttttgt caaaaaattc
     2101 tctcgctaat ttatttgctt ttttaatgtt attattttat gactttttat agtcactgaa
     2161 aagtttgcat ctgagtgaag tgaatgctat caaaatgtga ttctgtctga tgtactttca
     2221 caatctctct tcaattccat tttgaagtgc tttaaacccg aaaggttgag aaaaatgcga
     2281 gcgctcaaat atttgtattg tgttcgttga gtgacccaac aaaaagagga aactttattg
     2341 tgccgccaag aaaaaagtct cacgtacggt gcgcgcgatg ccgtatcaca actgagaaaa
     2401 aagacaaaac ttattgaaca gtgttccttt caaaaaatta ttttgtaaaa aatcgagttt
     2461 ttctccaaaa taacgcacgc aatcaaaaac aagaaaacta acgtgttctc ctgggcgtgg
     2521 aactccttat caagattacc attattgtct gtcagaattg atatttcaat tatttcatat
     2581 ctagacaaac ccgggtgtct tgcatttatt agtactgtga gtttacatac aaacgaggct
     2641 tactcagata acatagagtc acctctaatt agcaaaaacc catttctaat tagctatttc
     2701 gaccagttct aaagtcaccg taattagaaa tggtttttgt agcttttcta attagaaatg
     2761 accccgagat aagtgaaata tggcattttc tgggttttct agtaagaaat gatcccgcac
     2821 gtagtgattc cgcgttacaa gccaaattca aattaaaaaa gtatattaca caattcaagt
     2881 tacattattt tcaggaaaca atggcgaaga gcaaagcaga gattcaaaag gaggaaagag
     2941 acaacagata tgaacgaaga cttcttgaaa agagaaagag acatgaggtg agtaaccatg
     3001 ataataaata atcaagtaag ggctatagat ttataaattc tggagttaac agtttcgaaa
     3061 aaaagtagtt ttcatatgtg cgaaggaata tcaaaaatag gggaatagcc taaacgagtg
     3121 agaaaattaa atgtttgtta ttttttcaaa aatgtctgca gggccgttat cctttcaaat
     3181 gcactctatc aaacatagat catatccatc atatcttatc cagaatttat atcatatcca
     3241 agattgagag cgtttcaagg aaaatctcaa acaaccatac aattttgtag atcgacaaca
     3301 aggcctccac ttcttcatcg atggcctctg cagcacctgc accaccaagt ttgtattttt
     3361 catattaata taaagtgagc cacgcatagt cgtttttagg atttttgcca gatcgatgtc
     3421 gcgcaaacag actgtttttt tttctttgaa agccgtgcgt gacatgggcc acggattgta
     3481 acgccgcgct ggatggataa aaatttattc agttttttag cctttacaaa atatggttcg
     3541 actactttct ttatttttac atttttatag gtgctcattg agctgagata ttttgggtat
     3601 aatcggaaaa tcgctgattt tggtcatttt tctgcactaa agattacaca aaaagttcag
     3661 tatttagatt cattgttgaa aattggggat gatctacaga tacatatcgc cgtgttcaaa
     3721 aacacacgtg acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3781 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3841 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     3901 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     3961 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4021 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4081 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4141 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4201 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4261 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4321 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4381 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4441 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4501 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4561 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4621 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4681 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4741 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4801 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4861 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4921 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4981 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5041 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5101 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5161 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5221 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5281 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5341 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5401 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5461 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5521 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5581 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5641 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5701 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5761 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5821 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5881 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5941 cggccttttt acggttcctg gtcttttgct ggccttttgc tcacatgttc tttcctgcgt
     6001 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accagcggat
     6061 aacaatttca cagggccctg tcggattatg ggctcttctt ttatcctcta ctttctttct
     6121 agaaaatgca aatgttgagt cagaaattgc tgaatggagt acaactgcaa tgggtttagc
     6181 ttatgtttct ggaacaatta ctggtggaat tgtgattgaa aagtaaggtt atatttcatt
     6241 tcaataaatg ttaccacagt ataacaaaaa caagttacat tgttcagagt atgtctgaat
     6301 gtattattat tattgtattt gtgcatcagt actgtagttg tactgaagaa gggtttaaaa
     6361 taatctcaac gtttttaaga cgagttagtg agaatacggt cgcagttagg atgtttggac
     6421 atagttttat tagataatcg tgaactaact cactataaaa acccgctaac tcattgaata
     6481 agatagtttt aaacgttcaa aatttgaaga caaaattgta tgaataagtt tacagagttg
     6541 gtcgacgcaa gcttctcctt ctcttcacat ttctcaataa tttagctcta attgcattcg
     6601 tattcttcgc aaaaatacgt attctgattg acccgatgaa atacggatgc ctagttgcat
     6661 tgatcatcta cggttatact tatgggtgag gtttttcaaa tatattttga tttaatcgaa
     6721 taatgaaatt tcagaactgg agttggtcca atttcgtggt ttatatcctc tgaactggta
     6781 cctcaaaaac atcgtagtat tgctcaatct gtggcatatt caatgaatac tgtaatggtg
     6841 gttatcagta catttacagt attacctcta tattccgtta ttggtagtta tgctttcctt
     6901 attctatatt caattccttc atttataagt atgcttatat tatttcgcta cttaccggaa
     6961 accaaaggac gagagattca tgatattgtg aatgaactta aaaaaaaatg aaaacctttg
     7021 aaaacgttca tacggccgaa attttacttg atcgagccaa ttgaactttt gttaagactt
     7081 acagaaaagt gagagatttt gttacaaaaa agtgagaaaa aagcgaaaat gaacaggaaa
     7141 aagaagaaaa atacacccga agtatcatat aaaaattaaa aagattgttt gacctggcgg
     7201 aacttttttg caaaaactct gaatatccat ggcacatgct tcgtgcaaaa caggatccaa
     7261 atgaatatca acaagagatt cttgaagacg acgagccact tgctcagagc attgcttatc
     7321 tcgaatgaga cccttgttaa aatcattttt caaacattat atgacattgt cgaaatgtcc
     7381 tcctgattcc atgatggtag caaactcact tcgtctttga gccaatttat ccaagtcctt
     7441 gtaaaaagta ttcgaaaatt gttaacggat aaaatgttta ttataatatc aaaaacaatt
     7501 tgtcagttga ccactttttg atatatttga cagaaaccgg gatgaattgg ctcaaagtta
     7561 gggcgccctt ctattacagg gtttctgata acaaaccggt tattaactcc caacaaggga
     7621 tgatttcaat tcatcatgct caaattgacc caaattaagt tacatgacaa attcatcgcc
     7681 cttttcaact cttcttggct catcatctgt tattctgttc ctattctgtc tgcaccctat
     7741 acccctttgc atactctctc gtcattcctt tcttggatag tgtcttcttc tccccagctc
     7801 tgctacttct atgacttgcg cccgctgctt ttccgctcgt tcctctcctc tctgacgtct
     7861 tcttctgctt cttttctcct tcctttttcc cttctcaatc ttcctctttt tcccatttct
     7921 gtcaatcatt cgaagaagaa gaagaagaag accctcattc atttcatttt tttttctgtc
     7981 ggtgtgtgct gctcggttaa gagtgagctc tctattccac gtcttcttct ttttcttctt
     8041 cgattcgaat caatcactcc accaaaacgc attcgttttt gggattcacc ccgcggttcg
     8101 caataggttt ctttttcaaa ttattagcgt tataaataga aaaatggtgg agtttcaata
     8161 aaaatgataa tttacaaagt gtattttgat tacatgtact caaaaaggtt gaaatttttc
     8221 ataccagttt tccggaaatc catctgatat cattatcgta tttttctttt taaaaatgtt
     8281 ttcaaaaaaa aacaaaatat agctggttat ttggcaccct ctaattacca ttttcctgtc
     8341 acaccacact cttttccttc ttccacttct ttccgttttc agccgcttcc aaccaaaccg
     8401 atatgaaagc cgagcaacaa cgatcgattc cacccggctc ggcgaccttc ccgtcgcagg
     8461 tgagactcag aaaactagag aaaccgctca actaactctt gatatccgat ttcattcttt
     8521 tctcttttct ttttgttgaa cttttccata tttccagatg ccacggccac caccaagcac
     8581 cgaacaagga atcacaacgg aatcggagct tgcgaagaaa gctcaaatca ctccgaacga
     8641 cgttttagca cttccgggaa tcactcaagg tatcttgctc cctttggatt tcagtttaaa
     8701 acataaattt aggattctta tgctccccat ctgcgaacat ctataacatc gagttcacca
     8761 agttccaaat ccgtgatctg gacactgagc aagtgctttt cgagatcgcc aaaccggaga
     8821 acgatcagga gaatgatgag tcgccacagg agtcggcaag atacgtgcgt tatagatttg
     8881 ctccaaactt tttgaaactc aaaacggtcg gagcaactgt ggaattcaaa gtaggagaca
     8941 tcccaatcca tcatttccga atgatcgaac gtcacttctt caaagatcgc cttctgaagt
     9001 gttttgactt tgaattcgga ttctgtattc cgaattcacg aaacaactgt gaacatatct
     9061 atgagttccc tcaactctct caacaactca gtgagtcatt attctaaaaa gtacaattca
     9121 aaagactaat ctctttttca gtggacgaca tgatcaacaa tccaaacgag actcgttctg
     9181 acagcttcta tttcgtcgat aacaaactcg tcatgcacaa caaagccgac tactcatatg
     9241 atgcataaat atttaataca aaaatgttct ggataattat tctgtcgaat agaaaaaaaa
     9301 ctccaaatgt gattaaattc caataattcc tgtctagttt gttccttccc cttccccttc
     9361 tcatgttcaa tgcattccta agcttttcag ttcccccctt gtttctatat tttttcgctg
     9421 tcctgtcaca ctcgctaaaa acactaatca cacggaaatc tgttttcaat aaaaactcca
     9481 actttaactc attttcaatt tcaactgaaa gattttttca ttagagaatg tctagaacta
     9541 ggcccgggct acgtaatacg actcacttaa ggccttgact agagggtacc agagctcacc
     9601 taggcaggaa cagctatgac catgattacg ccaagctat
//
