LOCUS       sequence-149410-        5376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TagBFP labeled human Rab4a (GTPase located on recycling 
            endosomal membranes)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5376)
  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 5376)
  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-Rab4a-c-Myc
FEATURES             Location/Qualifiers
     source          1..5376
                     /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             2002..2031
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    2164..2285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2201..2220)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2255..2274
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2292..2747)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2429..2450)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2641..2660
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2774..2878
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2880..3237
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2880..3071
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2903..2923)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3088..3223
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3272..4066
                     /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             3272..4066
                     /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(3326..3345)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3936..3955
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4254..4273)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4298..4345
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4674..5262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5163..5182
                     /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 tgtcgcagac ggccatgtcc gaaacctacg attttttgtt taagttcttg
     1381 gttattggaa atgcaggaac tggcaaatct tgcttacttc atcagtttat tgaaaaaaaa
     1441 ttcaaagatg actcaaatca tacaatagga gtggaatttg gttcaaagat aataaatgtt
     1501 ggtggtaaat atgtaaagtt acaaatatgg gatacagcag gacaagaacg attcaggtcc
     1561 gtgacgagaa gttattaccg aggcgcggcc ggggctctcc tcgtctatga tatcaccagc
     1621 cgagaaacct acaatgcgct tactaattgg ttaacagatg cccgaatgct agcgagccag
     1681 aacattgtga tcatcctttg tggaaacaag aaggacctgg atgcagatcg tgaagttacc
     1741 ttcttagaag cctccagatt tgctcaagaa aatgagctga tgtttttgga aacaagtgcg
     1801 ctcacagggg agaatgtaga agaggctttt gtacagtgtg caagaaaaat acttaacaaa
     1861 atcgaatcag gtgagctgga cccagaaaga atgggctcag gtattcagta cggagatgct
     1921 gccttgagac agctgaggtc accgcggcgc gcacaggccc cgaacgctca ggagtgtggt
     1981 tgtacgcgta cgcggccgct cgagcagaaa ctcatctcag aagaggatct gtaaggatcc
     2041 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     2101 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2161 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2221 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2281 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2341 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2401 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2461 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2521 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2581 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2641 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2701 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     2761 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2821 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     2881 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3001 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3061 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3121 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3181 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3241 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3301 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3361 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3421 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     3481 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3541 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3601 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3661 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3721 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3781 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3841 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3901 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3961 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4021 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4081 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4141 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4201 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4261 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4321 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4381 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4441 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4501 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     4561 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4621 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4681 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4741 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4801 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4861 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4921 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4981 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5041 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5101 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5161 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5221 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5281 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5341 cctgattctg tggataaccg tattaccgcc atgcat
//
