LOCUS       sequence-160502-        8352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing SLC35B2-p2A-E2Crimson.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8352)
  AUTHORS   Park RJ, Wang T, Koundakjian D, Hultquist JF, Lamothe-Molina P, 
            Monel B, Schumann K, Yu H, Krupzcak KM, Garcia-Beltran W, 
            Piechocka-Trocha A, Krogan NJ, Marson A, Sabatini DM, Lander ES, 
            Hacohen N, Walker BD
  TITLE     A genome-wide CRISPR screen identifies a restricted set of HIV host 
            dependency factors.
  JOURNAL   Nat Genet. 2016 Dec 19. doi: 10.1038/ng.3741.
  PUBMED    27992415
REFERENCE   2  (bases 1 to 8352)
  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: "Nat Genet. 
            2016 Dec 19. doi: 10.1038/ng.3741."
COMMENT     Sequence Label: SLC35B2-p2A-E2Crimson
FEATURES             Location/Qualifiers
     source          1..8352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..212
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      236..245
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1559..1615
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1619..2290
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="STENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAKLQ
                     VTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGVVT
                     VTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHMAL
                     KLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHHLF
                     L"
     primer_bind     2011..2030
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2348..2381)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2437..3025
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2490..2510)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2908..2919)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3028..3044)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             3554..3734
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3796..4384)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3876..3895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4555..5346,5347..5415))
                     /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     5178..5197
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5416..5520)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5595..5614)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5786..6165
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6167..6365
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6320..6340
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             6383..6563
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6610..6735
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7232..7465
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7650..7694
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7992..8109
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     8167..8183
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8225..8258)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
       61 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
      121 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
      181 tttttcgcaa cgggtttgcc gccagaacac aggtgtcgtg acgcgggtta attaagccac
      241 catggacgct cgatggtggg ccgtagtggt gctggcagcc ttcccatcct tgggagcagg
      301 aggagaaacg cccgaggccc cccccgagag ttggacgcaa ctttggttct tccggttcgt
      361 cgtaaatgca gccggttacg ccagcttcat ggttccaggt tatctgctcg tgcagtattt
      421 caggagaaaa aactacctgg agacggggcg gggactgtgt tttccccttg tgaaagcttg
      481 cgtgttcggg aacgagccaa aagcctccga cgaggtccct ttggctcccc gaacggaggc
      541 cgcagagaca acacccatgt ggcaggcact gaaactcctt ttttgtgcta cgggattgca
      601 ggtgagttac ctgacatggg gggtccttca ggaacgcgtc atgactcggt cctacggcgc
      661 caccgccacc tcccccggag agcggttcac agactctcaa ttcctcgtgc tcatgaatcg
      721 ggtgctggcg ctgatcgtcg ccggtctgtc atgtgtgctg tgtaaacaac ccaggcatgg
      781 agccccgatg taccgctata gtttcgcatc actgagcaac gtactgtcta gctggtgtca
      841 gtacgaggct cttaaatttg tcagctttcc tacccaagtg ctcgccaaag ccagtaaagt
      901 cattccggtg atgctgatgg ggaaactggt atcccggcgc tcatacgagc attgggaata
      961 cctgacggcg acccttatca gtattggagt ctcaatgttt ttgctgtcaa gtgggcccga
     1021 accaaggagc agccctgcaa caacccttag cggactgatc ctgctggccg gctatatcgc
     1081 cttcgattct tttacaagta attggcagga tgccttgttc gcctacaaga tgtcctctgt
     1141 gcagatgatg tttggggtga atttctttag ctgcttgttc accgtgggca gtcttctgga
     1201 acaaggggcg ctgcttgagg gaacaagatt tatgggcaga cattccgagt ttgccgctca
     1261 cgctctgctg ctgagtattt gcagtgcttg cgggcagctc ttcatctttt acaccatcgg
     1321 tcaatttggg gccgctgtgt tcacaataat tatgactctg aggcaggcat tcgctatcct
     1381 cctcagctgt ctcctctacg gccataccgt aactgtcgta ggaggtctgg gtgtggccgt
     1441 tgtgttcgca gcactcctgc tgcgcgtata cgctagggga aggcttaagc aaagaggaaa
     1501 aaaagccgtc ccggtggaaa gtcctgtgca gaaggtggcg gccgcaggag gtggaggagc
     1561 tacaaatttc agcctgctga agcaggccgg cgacgtggag gagaaccccg gccccaatag
     1621 cactgagaac gtcatcaagc ccttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     1681 cggccacgag ttcgagatcg agggcgtggg cgagggcaag ccctacgagg gcacccagac
     1741 cgccaagctg caagtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc
     1801 ccagttcttc tacggctcca aggcgtacat caagcacccc gccgacatcc ccgactacct
     1861 caagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     1921 cgtggtgacc gtgacccagg actcctccct gcaggacggc accctcatct accacgtgaa
     1981 gttcatcggc gtgaacttcc cctccgacgg ccccgtaatg cagaagaaga ctctgggctg
     2041 ggagccctcc actgagcgca actacccccg cgacggcgtg ctgaagggcg agaaccacat
     2101 ggcgctgaag ctgaagggcg gcggccacta cctgtgtgag ttcaagtcca tctacatggc
     2161 caagaagccc gtgaagctgc ccggctacca ctacgtggac tacaagctcg acatcacctc
     2221 ccacaacgag gactacaccg tggtggagca gtacgagcgc gccgaggccc gccaccacct
     2281 gttcctgtga tgtacaagta aagcggccgg ccgccagcac agtggtcgaa attcgtcgag
     2341 ggacctaata acttcgtata gcatacatta tacgaagtta tacatgttta agggttccgg
     2401 ttccactagg tacaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     2461 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2521 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2581 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2641 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2701 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2761 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2821 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     2881 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     2941 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3001 atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgatcgag acctagaaaa
     3061 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
     3121 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
     3181 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     3241 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
     3301 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt
     3361 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg
     3421 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     3481 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca
     3541 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     3601 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     3661 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     3721 gaaaatctct agcagcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3781 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3841 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3901 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3961 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4021 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4081 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4141 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4201 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4261 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4321 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4381 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4441 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4501 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4561 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4621 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4681 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4741 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4801 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4861 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4921 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4981 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5041 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5101 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5161 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5221 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5281 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5341 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5401 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5461 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5521 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     5581 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     5641 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     5701 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     5761 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     5821 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     5881 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     5941 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     6001 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     6061 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     6121 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     6181 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     6241 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     6301 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     6361 agcagcgcgt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     6421 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     6481 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     6541 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     6601 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     6661 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     6721 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     6781 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     6841 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     6901 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     6961 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     7021 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccggc
     7081 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     7141 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     7201 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     7261 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     7321 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     7381 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     7441 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     7501 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     7561 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     7621 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     7681 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     7741 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     7801 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     7861 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     7921 ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta
     7981 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     8041 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     8101 caaaattttc gggtttatta cagggacagc agagatccag tttggttagt accgggcccg
     8161 ctctatcgag gtcgacggta tcgataagct cgcttcacga gattccagca ggtcgaggga
     8221 cctaataact tcgtatagca tacattatac gaagttatat taagggttcc aagcttaagc
     8281 ggccggccgc tgaaagaccc cacctgtagg tttggcaagc tagcggctcg agtggctccg
     8341 gtgcccgtca gt
//
