LOCUS       sequence-404773-        5897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus helper plasmid coding for Rev.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5897)
  AUTHORS   Dadonaite B, Crawford KHD, Radford CE, Farrell AG, Yu TC, Hannon WW,
            Zhou P, Andrabi R, Burton DR, Liu L, Ho DD, Chu HY, Neher RA, Bloom 
            JD
  TITLE     A pseudovirus system enables deep mutational scanning of the full 
            SARS-CoV-2 spike.
  JOURNAL   Cell. 2023 Mar 16;186(6):1263-1278.e20. doi: 
            10.1016/j.cell.2023.02.001. Epub 2023 Feb 13.
  PUBMED    36868218
REFERENCE   2  (bases 1 to 5897)
  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: 
            "10.1016/j.cell.2023.02.001"; journalName: "Cell"; date: 
            "2023-03-16- 16"; volume: "186"; issue: "6"; pages: "1263-1278.e20"
COMMENT     Sequence Label: pRC-CMV-Rev1b
FEATURES             Location/Qualifiers
     source          1..5897
                     /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"
     CDS             994..1344
                     /codon_start=1
                     /gene="rev"
                     /product="human immunodeficiency virus 1 (HIV-1) Rev 
                     protein"
                     /label=Rev
                     /note="enhances nuclear export of unspliced and 
                     incompletely spliced viral mRNAs"
                     /translation="MAGRSGDSDEDLLKAVRLIKFLYQSNPPPNPEGTRQARRNRRRRW
                     RERQRQIHSISERILSTYLGRSAEPVPLQLPPLERLTLDCNEDCGTSGTQGVGSPQILV
                     ESPTVLESGTKE"
     promoter        complement(1356..1374)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1357..1374)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1394..1411)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1400..1624
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1670..2183
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1757..1776)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1967..1988
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2206..2226)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2211..2540
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2391..2526
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2607..3401
                     /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(2661..2680)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3271..3290
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3575..3709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3612..3631)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3666..3685
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3740..3756)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3740..3756)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3753..3775)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3764..3780
                     /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(3788..3794,3795..3812,3813..3818))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3833..3854
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3971..3988)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4142..4730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4222..4241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4901..5692,5693..5761))
                     /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     5524..5543
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5762..5866)
                     /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 gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttctgcagat atccatcaca
      961 ctggcggccg ctcgagcatg catctagact gccatggcag gaagaagcgg agacagcgac
     1021 gaagacctcc tcaaggcagt cagactcatc aagtttctct atcaaagcaa cccacctccc
     1081 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     1141 agacagatcc attcgattag tgaacggatc cttagcactt atctgggacg atctgcggag
     1201 cctgtgcctc ttcagctacc accgcttgag agacttactc ttgattgtaa cgaggattgt
     1261 ggaacttctg ggacgcaggg ggtgggaagc cctcaaatat tggtggaatc tcctacagta
     1321 ttggagtcag gaactaaaga ataggatccg gctattctat agtgtcacct aaatgctaga
     1381 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     1441 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     1501 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     1561 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     1621 atggcttctg aggcggaaag aaccagctgg ggctcgaggg gggatcccca cgcgccctgt
     1681 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1741 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     1801 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     1861 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     1921 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     1981 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2041 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2101 aacaaaatat taacgcttac aatttaaata tttgcttata caatcttcct gtttttgggg
     2161 cttttctgat tatcaaccgg ggtgggtacc gagctcgaat tctgtggaat gtgtgtcagt
     2221 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2281 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2341 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2401 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2461 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2521 gaggcctagg cttttgcaaa aagctcccgg gagcttggat atccattttc ggatctgatc
     2581 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     2641 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2701 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2761 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     2821 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2881 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     2941 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3001 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3061 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3121 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3181 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3241 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3301 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3361 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3421 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3481 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3541 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     3601 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3661 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gtcgacctcg
     3721 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3781 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3841 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3901 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3961 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4021 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4081 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4141 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4201 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4261 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4321 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4381 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4441 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4501 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4561 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4621 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4681 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4741 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4801 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4861 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4921 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4981 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5041 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5101 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5161 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5221 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5281 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5341 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5401 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5461 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5521 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5581 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5641 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5701 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5761 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5821 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5881 aagtgccacc tgacgtc
//
