LOCUS       sequence-232659-        5811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes codon optimized FLAG tagged SIVRCM-Vpx.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5811)
  AUTHORS   Yurkovetskiy L, Guney MH, Kim K, Goh SL, McCauley S, Dauphin A, 
            Diehl WE, Luban J
  TITLE     Primate immunodeficiency virus proteins Vpx and Vpr counteract 
            transcriptional repression of proviruses by the HUSH complex.
  JOURNAL   Nat Microbiol. 2018 Dec;3(12):1354-1361. doi: 
            10.1038/s41564-018-0256-x. Epub 2018 Oct 8.
  PUBMED    30297740
REFERENCE   2  (bases 1 to 5811)
  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: 
            "10.1038/s41564-018-0256-x"; journalName: "Nat Microbiol"; date: 
            "2018-12"; volume: "3"; issue: "12"; pages: "1354-1361"
COMMENT     Sequence Label: pcDNA3.1 FLAG-SIV-RCM-vpx
FEATURES             Location/Qualifiers
     source          1..5811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      921..930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             930..1001
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     primer_bind     complement(1402..1419)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1408..1632
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1678..2106
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1765..1784)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1975..1996
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2115..2135)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2120..2449
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2300..2435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2362..2381
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2516..3310
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2570..2589)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3180..3199
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3484..3605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3521..3540)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3575..3594
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3654..3670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3654..3670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3667..3689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3678..3694
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3702..3708,3709..3726,3727..3732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3747..3768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3885..3902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4056..4644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4136..4155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4815..5606,5607..5675))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5438..5457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5676..5780)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga gccgccatgg attataaaga tgacgatgac aaagactata
      961 aagacgatga cgacaaagat tacaaggacg acgacgacaa agcggccgct atggcagaac
     1021 gggcccctga ggtgcctacc ggcgcgggag aagttgagtt tcaaccgtgg ctggcacgaa
     1081 tgctctatga aatcaatcaa gaggccaggc tgcactttca tcctgagttt atctttagac
     1141 tttggcgcac ttgtgtagaa cactggcatg acgagttggg tcgctccctc gaatacgctg
     1201 ggtatagata cctcctcttg atgcaaaagg ccctcttcac tcacatgcgc agcggctgca
     1261 gattgcggca cggagatcct cccagacaga gaggcgagcg ggttccgatt ctgcctggca
     1321 tgcagtaacc cggggaattc caccacactg gactagtgga tccgagctcg gtaccaagct
     1381 taagtttaaa ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     1441 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     1501 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     1561 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     1621 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     1681 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1741 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1801 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1861 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1921 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1981 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2041 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2101 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2161 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2221 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2281 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2341 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     2401 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     2461 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     2521 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     2581 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     2641 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     2701 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     2761 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     2821 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     2881 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     2941 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3001 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3061 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3121 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3181 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3241 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     3301 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     3361 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     3421 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     3481 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3541 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3601 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     3661 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3721 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3781 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3841 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3901 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3961 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4021 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4081 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4141 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4201 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4261 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4321 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4381 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4441 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4501 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4561 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4621 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4681 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4741 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4801 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4861 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4921 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4981 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5041 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5101 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5161 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5221 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5281 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5341 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5401 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5461 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5521 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5581 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5641 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5701 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5761 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
