LOCUS       sequence-454375-        5340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human Rab4B fluorescently tagged with GFP on the N-terminus for 
            mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5340)
  AUTHORS   Rowland AA, Chitwood PJ, Phillips MJ, Voeltz GK
  TITLE     ER Contact Sites Define the Position and Timing of Endosome Fission.
  JOURNAL   Cell. 2014 Nov 20;159(5):1027-41. doi: 10.1016/j.cell.2014.10.023.
  PUBMED    25416943
REFERENCE   2  (bases 1 to 5340)
  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.cell.2014.10"; journalName: "Cell"; date: "2014-11-20- 
            20"; volume: "159"; issue: "5"; pages: "1027-41"
COMMENT     Sequence Label: GFP-Rab4B
FEATURES             Location/Qualifiers
     source          1..5340
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(353..1069)
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     regulatory      1066..1075
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1114..1317)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1143..1163)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1318..1621
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1796..2384)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1876..1895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2713..2760
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2785..2804
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2992..3786)
                     /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             complement(2992..3786)
                     /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(3103..3122)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3713..3732
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3821..4178)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3835..3970
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3987..4178
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4135..4155
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4180..4284)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4311..4766
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4398..4417)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4608..4629
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4773..4894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4784..4803)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4838..4857
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 tgttggggct ggccagggtg cgggcatccg tcagccaggc agccagtgag ttgtatgtct
       61 cccggctggt gatgtcgtac accagcaggg ctccagccgc ccctcggtaa taactccgcg
      121 tcactgaccg aaaccgctcc tggccagccg tgtcccaaat ctgtagcttc acagtcttcc
      181 cacccacgtt gaccacccgg gatccaaact ccacgccgat tgtgtggttg gagtcctgtt
      241 tgaacttatt ctcaatgaac tgatgaagga gacatgattt gccagttcct gcactgccaa
      301 tcaccaggaa tttgaagagg aagtcgtagg tctcagcaga tctgagtccg gacttgtaca
      361 gctcatccat gccgtgggtg atggcggcgg cggtcacgaa gccgaagtag atcatgtgat
      421 cgcgcttctc gttggggtcc ttggacaggg cgctctgggt ggacaggtag tggttatcgg
      481 gcagcagcac agggccatcg ccgatggggg tattctgctg gtagtggtcg gccagctgca
      541 cgctgccatc ctcgatgttg tggcggatct tgaagttcac cttgatgcca ttcttggcct
      601 tgtcggtcat gatgtacaca ttgtgggcgt tgtagttgta ctccatctta ttgcccagga
      661 tgttgccatc ctccttgaaa tcggtgccgg tcagctcgat gcgattcacc agggtatcgc
      721 cctcgaactt cacctcggcg cgcgacttgt agttgccgtc atcctcgaag aagatggtgc
      781 gctcctggat gtagccctca ggcatggcgc tcttgaagaa gtcgtgctgc ttcatgtgat
      841 cggggtagcg tgagaagcac tgcacgccgt agctcagggt ggtcaccagg gtgggccagg
      901 gcacaggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
      961 cgccctcgcc ctcgccgctc acgctgaact tgtggccatt cacatcgcca ttcagctcga
     1021 tcaggatggg cacgatgccg gtgaacagct cggcgccctt gctcaccatg gtggcgaccg
     1081 gtagcgctag cggatctgac ggttcactaa accagctctg cttatataga cctcccaccg
     1141 tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc
     1201 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
     1261 tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat
     1321 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat
     1381 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact
     1441 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     1501 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     1561 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     1621 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact attaataact
     1681 aatgcatggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1741 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1801 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1861 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1921 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1981 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2041 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2101 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2161 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2221 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2281 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2341 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2401 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2461 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2521 tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg
     2581 cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg
     2641 cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa
     2701 ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg
     2761 tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag
     2821 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     2881 ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg
     2941 atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa
     3001 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag
     3061 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa
     3121 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa
     3181 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc
     3241 ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg
     3301 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg
     3361 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag
     3421 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag
     3481 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac
     3541 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc
     3601 gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc
     3661 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc
     3721 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc
     3781 aatcatgcga aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc
     3841 caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc tcggcctctg
     3901 cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta
     3961 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
     4021 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
     4081 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     4141 actgacacac attccacagc tggttctttc cgcctcagga ctcttccttt ttcaatatta
     4201 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4261 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg
     4321 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4381 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4441 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4501 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4561 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4621 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4681 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4741 taacaaaata ttaacgctta caatttacgc gttaagatac attgatgagt ttggacaaac
     4801 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4861 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4921 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     4981 tggtatggct gattatgatc agttatctag atccggtgga tcccgggccc gcggtacctc
     5041 agcagccaca cggctgaggg gccacggcct gggcactccg aggctgccga agctggcgga
     5101 gggacgcatc cccgtactga atgccagagc ccatcctctc cgggtctagc tcgcctgagt
     5161 caatcttgtt gaggatagtg cgggcacact tgaggaacgc ctcctccacg ttctcgcctg
     5221 tgagagcgct ggtctccagg aacatcagct cattctcctg ggcaaagcgg gaggcctcca
     5281 ggaaagtgac ctcccgctca gggtccaggt ccttcttgtt gccacagagg atgaccacga
//
