LOCUS       sequence-399275-        6221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses fluorescent protein-tagged enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6221)
  AUTHORS   Wills RC, Doyle CP, Zewe JP, Pacheco J, Hansen SD, Hammond GRV
  TITLE     A novel homeostatic mechanism tunes PI(4,5)P2-dependent signaling at
            the plasma membrane.
  JOURNAL   J Cell Sci. 2023 Aug 3:jcs.261494. doi: 10.1242/jcs.261494.
  PUBMED    37534432
REFERENCE   2  (bases 1 to 6221)
  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: "J Cell Sci.
            2023 Aug 3:jcs.261494. doi: 10.1242/jcs.261494."
COMMENT     Sequence Label: iRFP670-PIP4K2C
FEATURES             Location/Qualifiers
     source          1..6221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             102..896
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             102..896
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(156..175)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     766..785
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1084..1103)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1128..1175
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1504..2092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1993..2012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2267..2570
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2571..2774
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2725..2745
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2813..2822
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2819..3763
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     polyA_signal    5215..5336
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5252..5271)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5306..5325
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5343..5798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5480..5501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5692..5711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5825..5929
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5931..6221,1..67)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5931..6122
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5954..5974)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6139..6221,1..53)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
       61 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
      121 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
      181 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
      241 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
      301 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
      361 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
      421 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
      481 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
      541 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
      601 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
      661 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
      721 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
      781 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
      841 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
      901 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
      961 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     1021 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     1081 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     1141 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     1201 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     1261 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     1321 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     1381 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1441 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1501 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1561 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1621 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     1681 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1741 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     1801 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     1861 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     1921 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     1981 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2041 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2101 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2161 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     2221 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     2281 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     2341 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     2401 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     2461 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     2521 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     2581 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     2641 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     2701 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     2761 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat
     2821 ggtagcaggt catgcctctg gcagccccgc attcgggacc gcctctcatt cgaattgcga
     2881 acatgaagag atccacctcg ccggctcgat ccagccgcat ggcgcgcttc tggtcgtcag
     2941 cgaacatgat catcgcgtca tccaggccag cgccaacgcc gcggaatttc tgaatctcgg
     3001 aagcgtactc ggcgttccgc tcgccgagat cgacggcgat ctgttgatca agatcctgcc
     3061 gcatctcgat cccaccgccg aaggcatgcc ggtcgcggtg cgctgccgga tcggcaatcc
     3121 ctctacggag tactgcggtc tgatgcatcg gcctccggaa ggcgggctga tcatcgaact
     3181 cgaacgtgcc ggcccgtcga tcgatctgtc aggcacgctg gcgccggcgc tggagcggat
     3241 ccgcacggcg ggttcactgc gcgcgctgtg cgatgacacc gtgctgctgt ttcagcagtg
     3301 caccggctac gaccgggtga tggtgtatcg tttcgatgag caaggccacg gcctggtatt
     3361 ctccgagtgc catgtgcctg ggctcgaatc ctatttcggc aaccgctatc cgtcgtcgac
     3421 tgtcccgcag atggcgcggc agctgtacgt gcggcagcgc gtccgcgtgc tggtcgacgt
     3481 cacctatcag ccggtgccgc tggagccgcg gctgtcgccg ctgaccgggc gcgatctcga
     3541 catgtcgggc tgcttcctgc gctcgatgtc gccgtgccat ctgcagttcc tgaaggacat
     3601 gggcgtgcgc gccaccctgg cggtgtcgct ggtggtcggc ggcaagctgt ggggcctggt
     3661 tgtctgtcac cattatctgc cgcgcttcat ccgtttcgag ctgcgggcga tctgcaaacg
     3721 gctcgccgaa aggatcgcga cgcggatcac cgcgcttgag agctccggac tcagatctcg
     3781 agctcaagct tcgaattctg cagtcgacgg taccgcgatg gcgtcctcct cggtcccacc
     3841 agccacggta tcggcggcga cagcaggccc cggcccaggt ttcggcttcg cctccaagac
     3901 caagaagaag catttcgtgc agcagaaggt gaaggtgttc cgggcggccg acccgctggt
     3961 gggtgtgttc ctgtggggcg tagcccactc gatcaatgag ctcagccagg tgcctccccc
     4021 ggtgatgctg ctgccagatg actttaaggc cagctccaag atcaaggtca acaatcacct
     4081 tttccacagg gaaaatctgc ccagtcattt caagttcaag gagtattgtc cccaggtctt
     4141 caggaacctc cgtgatcgat ttggcattga tgaccaagat tacttggtgt cccttacccg
     4201 aaaccccccc agcgaaagtg aaggcagtga tggtcgcttc cttatctcct acgatcggac
     4261 tctggtcatc aaagaagtat ccagtgagga cattgctgac atgcatagca acctctccaa
     4321 ctatcaccag tacattgtga agtgccatgg caacacgctt ctgccccagt tcctggggat
     4381 gtaccgagtc agtgtggaca acgaagacag ctacatgctt gtgatgcgca atatgtttag
     4441 ccaccgtctt cctgtgcaca ggaagtatga cctcaagggt tccctagtgt cccgggaagc
     4501 cagcgataag gaaaaggtta aagaattgcc cacccttaag gatatggact ttctcaacaa
     4561 gaaccagaaa gtatatattg gtgaagagga gaagaaaata tttctggaga agctgaagag
     4621 agatgtggag tttctagtgc agctgaagat catggactac agccttctgc taggcatcca
     4681 cgacatcatt cggggctctg aaccagagga ggaagcgccc gtgcgggagg atgagtcaga
     4741 ggtggatggg gactgcagcc tgactggacc tcctgctctg gtgggctcct atggcacctc
     4801 cccagagggt atcggaggct acatccattc ccatcggccc ctgggcccag gagagtttga
     4861 gtccttcatt gatgtctatg ccatccggag tgctgaagga gccccccaga aggaggtcta
     4921 cttcatgggc ctcattgata tccttacaca gtatgatgcc aagaagaaag cagctcatgc
     4981 agccaaaact gtcaagcatg gggctggggc agagatctct actgtccatc cggagcagta
     5041 tgctaagcga ttcctggatt ttattaccaa catctttgcc taacgggatc caccggatct
     5101 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     5161 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     5221 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5281 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     5341 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     5401 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     5461 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     5521 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     5581 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5641 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5701 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5761 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     5821 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5881 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     5941 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     6001 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     6061 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     6121 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6181 catggctgac taattttttt tatttatgca gaggccgagg c
//
