LOCUS       sequence-210323-        4991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for HIV-1 VPR fused with TEV protease (TEVp).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4991)
  AUTHORS   Jones DM, Padilla-Parra S
  TITLE     Imaging real-time HIV-1 virion fusion with FRET-based biosensors.
  JOURNAL   Sci Rep. 2015 Aug 24;5:13449. doi: 10.1038/srep13449.
  PUBMED    26300212
REFERENCE   2  (bases 1 to 4991)
  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 Rep."; 
            date: "2015-08-24"; volume: "5:13449. doi"; pages: " 
            10.1038/srep13449"
COMMENT     Sequence Label: TEV-VPR
FEATURES             Location/Qualifiers
     source          1..4991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        146..449
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        450..653
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     604..624
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             685..1392
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
                     EPFQPVKEATQLMN"
     polyA_signal    1864..1985
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1901..1920)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1955..1974
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1992..2447)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2129..2150)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2341..2360
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2474..2578
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2580..2937
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2580..2771
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2603..2623)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2788..2923
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2972..3766
                     /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             2972..3766
                     /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(3026..3045)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3636..3655
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3954..3973)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3998..4045
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4374..4962
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4863..4882
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
       61 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
      121 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      181 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      241 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      301 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      361 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      421 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      481 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      541 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      601 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      661 tgaaccgtca gatccgctag catgggagaa agcttgttta aggggccgcg tgattacaac
      721 ccgatatcga gcaccatttg tcatttgacg aatgaatctg atgggcacac aacatcgttg
      781 tatggtattg gatttggtcc cttcatcatt acaaacaagc acttgtttag aagaaataat
      841 ggaacactgt tggtccaatc actacatggt gtattcaagg tcaagaacac cacgactttg
      901 caacaacacc tcattgatgg gagggacatg ataattattc gcatgcctaa ggatttccca
      961 ccatttcctc aaaagctgaa atttagagag ccacaaaggg aagagcgcat atgtcttgtg
     1021 acaaccaact tccaaactaa gagcatgtct agcatggtgt cagacactag ttgcacattc
     1081 ccttcatctg atggcatatt ctggaagcat tggattcaaa ccaaggatgg gcagtgtggc
     1141 agtccattag tatcaactag agatgggttc attgttggta tacactcagc atcgaatttc
     1201 accaacacaa acaattattt cacaagcgtg ccgaaaaact tcatggaatt gttgacaaat
     1261 caggaggcgc agcagtgggt tagtggttgg cgattaaatg ctgactcagt attgtggggg
     1321 ggccataaag ttttcatggt gaaacctgaa gagccttttc agccagttaa ggaagcgact
     1381 caactcatga attactcaga tctcgagctc aagcttcgaa ttctgcagtc gacggtacca
     1441 gaacaagccc cagaagacca agggccacag agggagccac acaatgaatg gacactagag
     1501 cttttagagg agcttaagaa tgaagctgtt agacattttc ctaggatttg gctccatggc
     1561 ttagggcaac atatctatga aacttatggg gatacttggg caggagtgga agccataata
     1621 agaattctgc aacaactgct gtttatccat ttcagaattg ggtgtcgaca tagcagaata
     1681 ggcgttactc aacagaggag agcaagaaat ggagccagta gatcctagtc tagaggatcc
     1741 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     1801 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     1861 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1921 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1981 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2041 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2101 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2161 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2221 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2281 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2341 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2401 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     2461 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2521 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     2581 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2641 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     2701 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2761 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2821 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     2881 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     2941 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3001 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3061 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3121 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     3181 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3241 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3301 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3361 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3421 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3481 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3541 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3601 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3661 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3721 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3781 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     3841 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3901 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     3961 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4021 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4081 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4141 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4201 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     4261 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4321 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4381 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4441 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4501 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4561 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4621 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4681 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4741 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4801 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4861 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4921 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4981 tttttacggt t
//
