LOCUS       sequence-399526-        6644 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 6644)
  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 6644)
  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-PIP5K1A
FEATURES             Location/Qualifiers
     source          1..6644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             137..931
                     /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             137..931
                     /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(191..210)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     801..820
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1119..1138)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1163..1210
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1539..2127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2028..2047
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2302..2605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2606..2809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2760..2780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2848..2857
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2854..3798
                     /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    5673..5794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5710..5729)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5764..5783
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5801..6256)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5938..5959)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6150..6169
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6283..6387
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6389..6644,1..102)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6389..6580
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6412..6432)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6597..6644,1..88)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
       61 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
      121 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
      181 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
      241 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
      301 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
      361 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
      421 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
      481 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
      541 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
      601 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
      661 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
      721 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
      781 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
      841 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
      901 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
      961 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     1021 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     1081 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     1141 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     1201 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     1261 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     1321 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     1381 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     1441 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1501 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1561 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1621 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1681 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1741 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1801 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1861 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1921 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1981 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2041 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2101 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2161 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2221 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     2281 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2341 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2401 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2461 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2521 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2581 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     2641 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2701 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2761 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     2821 ccgtcagatc cgctagcgct accggtcgcc accatggtag caggtcatgc ctctggcagc
     2881 cccgcattcg ggaccgcctc tcattcgaat tgcgaacatg aagagatcca cctcgccggc
     2941 tcgatccagc cgcatggcgc gcttctggtc gtcagcgaac atgatcatcg cgtcatccag
     3001 gccagcgcca acgccgcgga atttctgaat ctcggaagcg tactcggcgt tccgctcgcc
     3061 gagatcgacg gcgatctgtt gatcaagatc ctgccgcatc tcgatcccac cgccgaaggc
     3121 atgccggtcg cggtgcgctg ccggatcggc aatccctcta cggagtactg cggtctgatg
     3181 catcggcctc cggaaggcgg gctgatcatc gaactcgaac gtgccggccc gtcgatcgat
     3241 ctgtcaggca cgctggcgcc ggcgctggag cggatccgca cggcgggttc actgcgcgcg
     3301 ctgtgcgatg acaccgtgct gctgtttcag cagtgcaccg gctacgaccg ggtgatggtg
     3361 tatcgtttcg atgagcaagg ccacggcctg gtattctccg agtgccatgt gcctgggctc
     3421 gaatcctatt tcggcaaccg ctatccgtcg tcgactgtcc cgcagatggc gcggcagctg
     3481 tacgtgcggc agcgcgtccg cgtgctggtc gacgtcacct atcagccggt gccgctggag
     3541 ccgcggctgt cgccgctgac cgggcgcgat ctcgacatgt cgggctgctt cctgcgctcg
     3601 atgtcgccgt gccatctgca gttcctgaag gacatgggcg tgcgcgccac cctggcggtg
     3661 tcgctggtgg tcggcggcaa gctgtggggc ctggttgtct gtcaccatta tctgccgcgc
     3721 ttcatccgtt tcgagctgcg ggcgatctgc aaacggctcg ccgaaaggat cgcgacgcgg
     3781 atcaccgcgc ttgagagctc cggactcaga tctcgagctc aagcttcgaa ttctgcagtc
     3841 gacggtaccg cgatggcgtc ggcctcctcc gggccgtcgt cttcggtcgg tttttcatcc
     3901 tttgatcccg cggtcccttc ctgtaccttg tcctcagcag catctggaat caagagaccc
     3961 atggcatctg aggtcttgga agctagacag gattcttaca tctcattggt gccttatgcc
     4021 tctggcatgc ccatcaagaa aataggccat agaagtgttg attcctcagg agagacaaca
     4081 tataaaaaga caacctcatc agccttgaaa ggtgccatcc agttaggcat tacccacact
     4141 gtggggagcc tgagtaccaa accagagcgt gatgtcctca tgcaagattt ctacgtggtt
     4201 gagagtatct tctttcccag tgaagggagc aacctgaccc ctgctcatca ctacaatgac
     4261 tttcgtttca agacctatgc acctgttgcc ttccgctact tccgggagct atttggtatc
     4321 cggcccgatg attacttgta ttccctctgc agtgagccgc tgattgaact ctgtagctct
     4381 ggagctagtg gttccctatt ctatgtgtcc agcgacgatg agttcattat taagacagtc
     4441 caacataaag aggcggaatt tctgcagaag ctgcttccag gatactacat gaacctcaac
     4501 cagaaccctc ggactttgct gcctaaattc tatggactgt actgtgtgca ggcaggtggc
     4561 aagaacattc ggattgtggt gatgaacaat cttttaccaa gatcggtaaa aatgcatatc
     4621 aaatatgacc tcaaaggctc aacctacaaa cggcgggctt cccagaaaga gcgagagaag
     4681 cctcttccca catttaaaga cctagacttc ttacaagaca tccctgatgg tctttttttg
     4741 gatgctgaca tgtacaacgc tctctgtaag accctgcagc gtgactgttt ggtgctgcag
     4801 agcttcaaga taatggatta cagcctcttg atgtcaatcc ataatataga tcatgcacaa
     4861 cgagagccct taagcagtga aacacagtac tcagttgata ctcgaagacc ggccccccaa
     4921 aaggctctgt attccacagc catggaatcc atccagggag aggctcgacg gggtggtacc
     4981 atggagactg atgaccatat gggtggcatc cctgcccgga atagtaaagg ggaaaggctt
     5041 ctgctttata ttggcatcat tgacattcta cagtcttaca ggtttgttaa gaagttggag
     5101 cactcttgga aagccctggt acatgacgga gacactgtct cagtgcatcg cccaggcttc
     5161 tacgctgaac ggttccagcg cttcatgtgc aacacagtat ttaagaagat tcccttgaag
     5221 ccttctcctt ccaaaaagtt tcggtctggc tcatctttct ctcggcgagc aggctccagt
     5281 ggcaactcct gcattactta ccagccatcg gtctctgggg aacacaaggc acaagtgaca
     5341 acaaaggcag aagtggagcc aggcgttcac cttggtcgtc ctgatgtttt acctcagact
     5401 ccacctttgg aggaaatcag tgagggctcg cctattcctg accccagttt ctcacctcta
     5461 gttggagaga ctttgcaaat gctaactaca agtacaacct tggaaaagct tgaagttgca
     5521 gagtcagagt tcacccatta acgggatcca ccggatctag ataactgatc ataatcagcc
     5581 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     5641 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     5701 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5761 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     5821 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     5881 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     5941 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     6001 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     6061 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     6121 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     6181 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     6241 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6301 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6361 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     6421 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     6481 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     6541 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     6601 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggc
//
