LOCUS       sequence-234223-        5832 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Encodes codon optimized FLAG tagged SIVMAC251-Vpx-Q76A mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5832)
  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 5832)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-MAC251-vpx-Q76A
FEATURES             Location/Qualifiers
     source          1..5832
                     /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(1423..1440)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1429..1653
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1699..2127
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1786..1805)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1996..2017
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2136..2156)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2141..2470
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2321..2456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2383..2402
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2537..3331
                     /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(2591..2610)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3201..3220
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3505..3626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3542..3561)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3596..3615
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3675..3691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3675..3691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3688..3710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3699..3715
                     /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(3723..3729,3730..3747,3748..3753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3768..3789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3906..3923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4077..4665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4157..4176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4836..5627,5628..5696))
                     /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     5459..5478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5697..5801)
                     /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 atgagcgatc
     1021 caagggaacg aatcccgccc ggaaattctg gtgaagagac catcggtgag gcgtttgagt
     1081 ggctgaaccg gactgtcgag gaaatcaaca gggaggctgt gaaccatctg cccagggaat
     1141 tgatatttca ggtatggcaa cgatcctggg agtattggca tgacgaacag ggcatgtcac
     1201 agagctacgt caagtatcgg tacttgtgtc ttatggctaa agctctcttc atgcattgca
     1261 aaaaggggtg ccgatgcttg ggagaaggtc atggtgcggg ggggtggcgc ccaggacccc
     1321 cacctcctcc accccctggg ttggcataac ccggggaatt ccaccacact ggactagtgg
     1381 atccgagctc ggtaccaagc ttaagtttaa accgctgatc agcctcgact gtgccttcta
     1441 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1501 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1561 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     1621 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     1681 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1741 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1801 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1861 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1921 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1981 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2041 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2101 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     2161 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2221 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2281 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2341 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2401 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2461 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     2521 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     2581 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     2641 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     2701 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2761 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     2821 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     2881 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     2941 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3001 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3061 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3121 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3181 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3241 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3301 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3361 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3421 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3481 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     3541 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3601 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     3661 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3721 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3781 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3841 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3901 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3961 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4021 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4081 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4141 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4201 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4261 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4321 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4381 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4441 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4501 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4561 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4621 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4681 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4741 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4801 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4861 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4921 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4981 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5041 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5101 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5161 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5221 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5281 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5341 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5401 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5461 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5521 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5581 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5641 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5701 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5761 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5821 ccacctgacg tc
//
