LOCUS       sequence-149412-        5367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TagBFP labeled human Rab5a (GTPase located on early 
            endosomal membranes)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5367)
  AUTHORS   Boothe T, Lim GE, Cen H, Skovso S, Piske M, Li SN, Nabi IR, Gilon P,
            Johnson JD
  TITLE     Inter-domain tagging implicates caveolin-1 in insulin receptor 
            trafficking and Erk signaling bias in pancreatic beta-cells.
  JOURNAL   Mol Metab. 2016 Feb 10;5(5):366-78. doi: 
            10.1016/j.molmet.2016.01.009. eCollection 2016 May.
  PUBMED    27110488
REFERENCE   2  (bases 1 to 5367)
  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: 
            "10.1016/j.molmet.2016.01.009"; journalName: "Mol Metab"; date: 
            "2016-02-10- 10"; volume: "5"; issue: "5"; pages: "366-78"
COMMENT     Sequence Label: pTag-BFP-C-h-Rab5a-c-Myc
FEATURES             Location/Qualifiers
     source          1..5367
                     /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"
     CDS             607..1305
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             1993..2022
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    2155..2276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2192..2211)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2246..2265
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2283..2738)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2420..2441)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2632..2651
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2765..2869
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2871..3228
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2871..3062
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2894..2914)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3079..3214
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3263..4057
                     /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             3263..4057
                     /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(3317..3336)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3927..3946
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4245..4264)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4289..4336
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4665..5253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5154..5173
                     /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 gctagcggtc
      601 gccaccatga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
      661 gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
      721 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
      781 gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac
      841 ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac
      901 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
      961 gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc
     1021 ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac
     1081 gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacatcaa gaccacatat
     1141 agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga
     1201 ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg
     1261 gccagatact gcgacctccc tagcaaactg gggcacaagc ttaattccgg actcagatct
     1321 cgagctcaaa tggctagtcg aggcgcaaca agacccaacg ggccaaatac tggaaataaa
     1381 atatgccagt tcaaactagt acttctggga gagtccgctg ttggcaaatc aagcctagtg
     1441 cttcgttttg tgaaaggcca atttcatgaa tttcaagaga gtaccattgg ggctgctttt
     1501 ctaacccaaa ctgtatgtct tgatgacact acagtaaagt ttgaaatatg ggatacagct
     1561 ggtcaagaac gataccatag cctagcacca atgtactaca gaggagcaca agcagccata
     1621 gttgtatatg atatcacaaa tgaggagtcc tttgcaagag caaaaaattg ggttaaagaa
     1681 cttcagaggc aagcaagtcc taacattgta atagctttat cgggaaacaa ggccgaccta
     1741 gcaaataaaa gagcagtaga tttccaggaa gcacagtcct atgcagatga caatagttta
     1801 ttattcatgg agacatccgc taaaacatca atgaatgtaa atgaaatatt catggcaata
     1861 gctaaaaaat tgccaaagaa tgaaccacaa aatccaggag caaattctgc cagaggaaga
     1921 ggagtagacc ttaccgaacc cacacaacca accaggaatc agtgttgtag taacacgcgt
     1981 acgcggccgc tcgagcagaa actcatctca gaagaggatc tgtaaggatc caccggatct
     2041 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2101 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2161 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2221 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     2281 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2341 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2401 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2461 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2521 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2581 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2641 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2701 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2761 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2821 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     2881 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3001 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3061 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3121 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3181 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3241 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3301 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3361 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3421 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3481 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3541 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3601 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3661 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3721 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3781 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3841 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3901 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3961 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4021 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4081 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4141 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4201 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4261 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4321 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4381 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4441 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4501 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4561 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4621 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4681 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4741 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4801 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4861 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4921 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4981 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5041 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5101 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5161 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5221 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5281 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5341 gtggataacc gtattaccgc catgcat
//
