LOCUS       sequence-78910-e        5931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruiter to Rab5-positive endosomes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5931)
  AUTHORS   Hammond GR, Machner MP, Balla T
  TITLE     A novel probe for phosphatidylinositol 4-phosphate reveals multiple 
            pools beyond the Golgi.
  JOURNAL   J Cell Biol. 2014 Apr 7.
  PUBMED    24711504
REFERENCE   2  (bases 1 to 5931)
  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 
            Biol. 2014 Apr 7."
COMMENT     Sequence Label: iRFP-FRB-Rab5
FEATURES             Location/Qualifiers
     source          1..5931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1560
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     CDS             1609..1887
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     polyA_signal    2719..2840
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2756..2775)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2810..2829
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2847..3302)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2984..3005)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3196..3215
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3329..3433
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3435..3792
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3435..3626
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3458..3478)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3643..3778
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3827..4621
                     /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             3827..4621
                     /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(3881..3900)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4491..4510
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4809..4828)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4853..4900
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5229..5817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5718..5737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggcgga aggatccgtc gccaggcagc ctgacctctt gacctgcgac
      661 gatgagccga tccatatccc cggtgccatc caaccgcatg gactgctgct cgccctcgcc
      721 gccgacatga cgatcgttgc cggcagcgac aaccttcccg aactcaccgg actggcgatc
      781 ggcgccctga tcggccgctc tgcggccgat gtcttcgact cggagacgca caaccgtctg
      841 acgatcgcct tggccgagcc cggggcggcc gtcggagcac cgatcactgt cggcttcacg
      901 atgcgaaagg acgcaggctt catcggctcc tggcatcgcc atgatcagct catcttcctc
      961 gagctcgagc ctccccagcg ggacgtcgcc gagccgcagg cgttcttccg ccgcaccaac
     1021 agcgccatcc gccgcctgca ggccgccgaa accttggaaa gcgcctgcgc cgccgcggcg
     1081 caagaggtgc ggaagattac cggcttcgat cgggtgatga tctatcgctt cgcctccgac
     1141 ttcagcggcg aagtgatcgc agaggatcgg tgcgccgagg tcgagtcaaa actaggcctg
     1201 cactatcctg cctcaaccgt gccggcgcag gcccgtcggc tctataccat caacccggta
     1261 cggatcattc ccgatatcaa ttatcggccg gtgccggtca ccccagacct caatccggtc
     1321 accgggcggc cgattgatct tagcttcgcc atcctgcgca gcgtctcgcc cgtccatctg
     1381 gaattcatgc gcaacatagg catgcacggc acgatgtcga tctcgatttt gcgcggcgag
     1441 cgactgtggg gattgatcgt ttgccatcac cgaacgccgt actacgtcga tctcgatggc
     1501 cgccaagcct gcgagctagt cgcccaggtt ctggcctggc agatcggcgt gatggaagag
     1561 tccggactca gatctcgagc tggaggtgct ggtgctatcc tatctagaat cctctggcat
     1621 gagatgtggc atgaaggcct ggaagaggca tctcgtttgt actttgggga aaggaacgtg
     1681 aaaggcatgt ttgaggtgct ggagcccttg catgctatga tggaacgggg cccccagact
     1741 ctgaaggaaa catcctttaa tcaggcctat ggtcgagatt taatggaggc ccaagagtgg
     1801 tgcaggaagt acatgaaatc agggaatgtc aaggacctca cccaagcctg ggacctctat
     1861 tatcatgtgt tccgacgaat ctcaaagggt ggtagtgctg gtggtagtgc tcaagcttcg
     1921 aattctgcag tcgacggtac catggctaat cgaggagcaa caagacccaa cgggccaaat
     1981 actggaaata aaatatgcca gttcaaacta gtacttctgg gagagtctgc tgttggcaaa
     2041 tcaagcctag tgcttcgttt tgtgaagggc caatttcatg aatttcaaga gagtaccata
     2101 ggggctgctt ttctaaccca aactgtgtgt cttgatgata caacagtaaa gtttgaaata
     2161 tgggatacag ctggtcaaga acgataccat agcttagcac caatgtacta cagaggagca
     2221 caagcagcca tagttgtata tgatatcaca aatgaggagt cctttgccag agccaaaaac
     2281 tgggttaaag aacttcagag gcaagccagt cctaacattg taatagcttt atcaggaaac
     2341 aaggctgatc ttgcaaataa aagagctgtc gatttccagg aagcacagtc ctatgcagat
     2401 gacaacagtt tattattcat ggagacatca gctaaaacat cgatgaacgt aaatgaaata
     2461 ttcatggcaa tagctaaaaa gttgccaaag aacgaaccac agaatccagg agcaaattct
     2521 gccagaggaa gaggagtaga ccttactgaa cccacgcagc caaccaggag tcagtgttgt
     2581 agtaactaag gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     2641 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2701 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2761 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2821 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     2881 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2941 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3001 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3061 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3121 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3181 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3241 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3301 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3361 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3421 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3481 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3541 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3601 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3661 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3721 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3781 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3841 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3901 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3961 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     4021 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4081 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4141 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4201 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4261 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4321 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4381 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4441 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4501 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4561 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4621 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4681 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4741 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     4801 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4861 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4921 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4981 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5041 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     5101 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5161 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5221 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5281 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5341 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5401 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5461 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5521 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5581 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5641 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5701 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5761 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5821 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5881 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
