LOCUS       sequence-309926-        5349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of endosome in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5349)
  AUTHORS   Lee J, Liu Z, Suzuki PH, Ahrens JF, Lai S, Lu X, Guan S, St-Pierre F
  TITLE     Versatile phenotype-activated cell sorting.
  JOURNAL   Sci Adv. 2020 Oct 23;6(43). pii: 6/43/eabb7438. doi: 
            10.1126/sciadv.abb7438. Print 2020 Oct.
  PUBMED    33097540
REFERENCE   2  (bases 1 to 5349)
  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: "Sci Adv."; 
            date: "2020-10-23"; volume: "6(43). pii: 6/43/eabb7438. doi"; pages:
            " 10.1126/sciadv.abb7438. Print 2020 Oct"
COMMENT     Sequence Label: pCMV-mGold-Rab4a-C-7
FEATURES             Location/Qualifiers
     source          1..5349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        151..454
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        455..658
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     609..629
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      697..706
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(703..705,706..708,709..1419)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(748..769)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1009..1028)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1356..1377
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2227..2348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2264..2283)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2318..2337
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2355..2810)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2492..2513)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2704..2723
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2837..2941
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2943..3300
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2943..3134
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2966..2986)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3151..3286
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3335..4129
                     /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             3335..4129
                     /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(3389..3408)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3999..4018
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4317..4336)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4361..4408
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4737..5325
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5226..5245
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
       61 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
      121 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      181 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      241 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      301 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      361 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      421 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      481 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      541 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      601 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
      661 tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca ccatggtgag caagggcgag
      721 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      781 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      841 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac cagcctgggc
      901 tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
      961 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1021 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1081 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1141 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     1201 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     1261 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     1321 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1381 gccgccggga tcactctcgg catggacgag ctgtacaagt ccggactcag atctcgagcc
     1441 atgtcgcaga cggccatgtc cgaaacctac gattttttgt ttaagttctt ggttattgga
     1501 aatgcaggaa ctggcaaatc ttgcttactt catcagttta ttgaaaaaaa attcaaagat
     1561 gactcaaatc atacaatagg agtggaattt ggttcaaaga taataaatgt tggtggtaaa
     1621 tatgtaaagt tacaaatatg ggatacagca ggacaagaac gattcaggtc cgtgacgaga
     1681 agttattacc gaggcgcggc cggggctctc ctcgtctatg atatcaccag ccgagaaacc
     1741 tacaatgcgc ttactaattg gttaacagat gcccgaatgc tggcgagcca gaacattgtg
     1801 atcatccttt gtggaaacaa gaaggacctg gatgcagatc gtgaagttac cttcttagaa
     1861 gcctccagat ttgctcaaga aaatgagctg atgtttttgg aaacaagtgc gctcacaggg
     1921 gagaatgtag aagaggcttt tgtacagtgt gcaagaaaaa tacttaacaa aatcgaatca
     1981 ggtgagctgg acccagaaag aatgggctca ggtattcagt acggagatgc tgccttgaga
     2041 cagctgaggt caccgcggcg cgcacaggcc ccgaacgctc aggagtgtgg ttgttaggga
     2101 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     2161 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2221 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2281 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2341 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2401 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2461 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2521 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2581 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2641 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2701 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     2761 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     2821 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2881 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2941 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3001 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3061 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3121 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3181 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3241 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3301 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3361 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3421 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3481 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     3541 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3601 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     3661 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     3721 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     3781 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     3841 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     3901 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     3961 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4021 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4081 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4141 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4201 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4261 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4321 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4381 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     4441 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     4501 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4561 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     4621 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4681 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4741 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4801 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4861 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4921 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4981 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5041 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5101 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5161 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5221 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5281 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5341 gccttttta
//
