LOCUS       sequence-223909-        6012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses Gag missing the p6 domain, tagged with mEGFP, in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6012)
  AUTHORS   Johnson DS, Bleck M, Simon SM
  TITLE     Timing of ESCRT-III protein recruitment and membrane scission during
            HIV-1 assembly.
  JOURNAL   Elife. 2018 Jul 4;7. pii: 36221. doi: 10.7554/eLife.36221.
  PUBMED    29972351
REFERENCE   2  (bases 1 to 6012)
  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: "Elife."; 
            date: "2018-07-4"; volume: "7. pii: 36221. doi"; pages: " 
            10.7554/eLife.36221"
COMMENT     Sequence Label: Gag-delta-p6-mEGFP
FEATURES             Location/Qualifiers
     source          1..6012
                     /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"
     regulatory      635..644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1952..1961
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1958..1960,1961..1963,1964..2677)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2003..2024)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2264..2283)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2611..2632
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2800..2921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2837..2856)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2891..2910
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2928..3383)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3065..3086)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3277..3296
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3410..3514
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3516..3873
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3516..3707
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3539..3559)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3724..3859
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3908..4702
                     /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             3908..4702
                     /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(3962..3981)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4572..4591
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4890..4909)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4934..4981
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5310..5898
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5799..5818
                     /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 gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcgccacc atgggcgccc gcgccagcgt
      661 gctgtcgggc ggcgagctgg accgctggga gaagatccgc ctgcgccccg gcggcaaaaa
      721 gaagtacaag ctgaagcaca tcgtgtgggc cagccgcgaa ctggagcgct tcgccgtgaa
      781 ccccgggctc ctggagacca gcgaggggtg ccgccagatc ctcggccaac tgcagcccag
      841 cctgcaaacc ggcagcgagg agctgcgcag cctgtacaac accgtggcca cgctgtactg
      901 cgtccaccag cgcatcgaaa tcaaggatac gaaagaggcc ctggataaaa tcgaagagga
      961 acagaataag agcaaaaaga aggcccaaca ggccgccgcg gacaccggac acagcaacca
     1021 ggtcagccag aactacccca tcgtgcagaa catccagggg cagatggtgc accaggccat
     1081 ctccccccgc acgctgaacg cctgggtgaa ggtggtggaa gagaaggctt ttagcccgga
     1141 ggtgataccc atgttctcag ccctgtcaga gggagccacc ccccaagatc tgaacaccat
     1201 gctcaacaca gtggggggac accaggccgc catgcagatg ctgaaggaga ccatcaatga
     1261 ggaggctgcc gaatgggatc gtgtgcatcc ggtgcatgca gggcccatcg caccgggcca
     1321 gatgcgtgag ccacggggct cagacatcgc cggaacgact agtacccttc aggaacagat
     1381 cggctggatg accaacaacc cacccatccc ggtgggagaa atctacaaac gctggatcat
     1441 cctgggcctg aacaagatcg tgcgcatgta tagccctacc agcatcctgg acatccgcca
     1501 aggcccgaag gaaccctttc gcgactacgt ggaccggttc tacaaaacgc tccgcgccga
     1561 gcaggctagc caggaggtga agaactggat gaccgaaacc ctgctggtcc agaacgcgaa
     1621 cccggactgc aagacgatcc tgaaggccct gggcccagcg gctaccctag aggaaatgat
     1681 gaccgcctgt cagggagtgg gcggacccgg ccacaaggca cgcgtcctgg ctgaggccat
     1741 gagccaggtg accaactccg ctaccatcat gatgcagcgc ggcaactttc ggaaccaacg
     1801 caagatcgtc aagtgcttca actgtggcaa agaagggcac acagcccgca actgcagggc
     1861 ccctaggaaa aagggctgtt ggaaatgtgg aaaggaagga caccaaatga aagattgtac
     1921 tgagagacag gctaattcgg atccaccggt cgccaccatg gtgagcaagg gcgaggagct
     1981 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2041 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2101 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     2161 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2221 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2281 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2341 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2401 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     2461 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2521 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     2581 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2641 cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata
     2701 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2761 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2821 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2881 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     2941 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3001 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3061 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3121 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3181 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3241 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3301 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3361 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3421 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3481 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3541 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3601 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3721 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3781 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3841 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3901 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3961 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4021 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4081 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4141 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4201 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4261 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4321 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4381 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4441 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4501 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4561 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4621 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4681 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4741 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4801 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4861 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4921 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4981 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5041 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5101 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5161 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5221 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5281 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5341 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5401 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5461 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5521 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5581 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5641 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5701 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5761 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5821 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ttcgtcaggg
     5881 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5941 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6001 accgccatgc at
//
