LOCUS       sequence-79078-e        5901 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruiter to Rab7-positive endosomes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5901)
  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 5901)
  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-Rab7
FEATURES             Location/Qualifiers
     source          1..5901
                     /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    2689..2810
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2726..2745)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2780..2799
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2817..3272)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2954..2975)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3166..3185
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3299..3403
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3405..3762
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3405..3596
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3428..3448)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3613..3748
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3797..4591
                     /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             3797..4591
                     /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(3851..3870)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4461..4480
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4779..4798)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4823..4870
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5199..5787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5688..5707
                     /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 atgacctcta ggaagaaagt gttgctgaag gttatcatcc tgggagattc tggagttggt
     1981 aagacatcac tcatgaacca gtatgtgaac aagaaattca gtaatcagta caaagctaca
     2041 ataggagcag actttctgac aaaggaggtg atggtggatg acagactagt tacaatgcag
     2101 atctgggaca cagcaggcca ggaacggttc cagtcccttg gtgtggcctt ctacagaggt
     2161 gcagactgct gcgttctggt atttgacgtt actgccccca acacattcaa aaccctcgat
     2221 agctggagag atgagtttct catccaggcc agtccccggg atcctgaaaa cttccctttc
     2281 gttgtgttgg gaaacaagat tgacctcgaa aacagacaag tggccacaaa gcgggcacag
     2341 gcctggtgct acagcaaaaa caacattccc tacttcgaga ccagtgccaa ggaggccatc
     2401 aatgtggagc aggcgttcca gacgattgca aggaatgcac ttaaacagga aacagaggtg
     2461 gagctgtaca atgaattccc tgaacccatc aaactggaca agaacgaccg ggccaagacc
     2521 tcagcggaaa gctgcagttg ctgaggtacc gcgggcccgg gatccaccgg atctagataa
     2581 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2641 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2701 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2761 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     2821 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2881 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2941 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3001 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3061 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3121 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3181 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3241 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3301 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3361 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     3421 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3481 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3541 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3601 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3661 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3721 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     3781 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3841 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3901 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3961 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4021 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4081 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4141 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4201 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4261 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4321 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4381 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4441 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4501 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4561 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4621 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4681 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4741 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     4801 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     4861 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4921 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4981 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5041 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5101 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5161 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5221 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5281 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5341 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5401 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5461 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5521 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5581 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5641 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5701 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5761 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5821 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5881 aaccgtatta ccgccatgca t
//
