LOCUS       sequence-265942-        5316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of TagBFP-Rab5 fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5316)
  AUTHORS   Friedman JR, Dibenedetto JR, West M, Rowland AA, Voeltz GK
  TITLE     Endoplasmic reticulum-endosome contact increases as endosomes 
            traffic and mature.
  JOURNAL   Mol Biol Cell. 2013 Apr;24(7):1030-40. doi: 10.1091/mbc.E12-10-0733.
            Epub 2013 Feb 6.
  PUBMED    23389631
REFERENCE   2  (bases 1 to 5316)
  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.E12-10-0733"; journalName: "Mol Biol Cell"; date: 
            "2013-04"; volume: "24"; issue: "7"; pages: "1030-40"
COMMENT     Sequence Label: BFP-Rab5
FEATURES             Location/Qualifiers
     source          1..5316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             547..1245
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    2044..2165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2081..2100)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2135..2154
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2172..2627)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2309..2330)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2521..2540
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2654..2758
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2760..3117
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2760..2951
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2783..2803)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2968..3103
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3152..3946
                     /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             3152..3946
                     /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(3206..3225)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3816..3835
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4134..4153)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4178..4225
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4554..5142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5043..5062
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcggtc
      541 gccaccatga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
      601 gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
      661 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
      721 gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac
      781 ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac
      841 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
      901 gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc
      961 ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac
     1021 gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacatcaa gaccacatat
     1081 agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga
     1141 ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg
     1201 gccagatact gcgacctccc tagcaaactg gggcacaagc ttaattccgg actcagatct
     1261 cgagctatga ctagcagaag cacagctagg cccaatgggc aaccccaggc cagcaaaatt
     1321 tgccagttca aattggtcct gctgggagaa tctgcagtgg gaaagtcaag cctggtatta
     1381 cgttttgtca aagggcagtt ccatgagtac caggagagca ccattggagc ggccttcctc
     1441 acccagtccg tttgtctaga tgacacaaca gtgaagtttg agatctggga cacagctggg
     1501 caggagcgat atcacagctt agcccccatg tactacaggg gtgcccaagc tgcaatcgtg
     1561 gtttacgaca ttactaatca ggaaaccttt gcccgagcaa agacatgggt gaaggaacta
     1621 cagcgacagg ccagtcctag catcgttatt gccctggcag ggaacaaagc tgacctggcc
     1681 aacaaacgta tggtggagta tgaagaggcc caggcatatg cagatgacaa cagcttattg
     1741 ttcatggaga cttcagccaa gacagctatg aacgtgaatg atctcttcct ggcaatagct
     1801 aagaagttgc caaagagtga accccagaat ctgggaggtg cagcaggccg aagccggggt
     1861 gtggatctcc atgaacagtc ccagcagaac aagagccagt gttgtagcaa ctgaggatcc
     1921 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     1981 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2041 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2101 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2161 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2221 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2281 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2341 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2401 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2461 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2521 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2581 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     2641 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2701 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     2761 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2821 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     2881 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2941 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3001 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3061 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3121 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3181 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3241 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3301 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     3361 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3421 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3481 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3541 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3601 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3661 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3721 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3781 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3841 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3901 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3961 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4021 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4081 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4141 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4201 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4261 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4321 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4381 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     4441 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4501 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4561 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4621 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4681 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4741 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4801 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4861 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4921 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4981 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5041 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5101 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5161 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5221 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     5281 ggggtcatta gttcatagcc catatatgga gttccg
//
