LOCUS       sequence-494097-        7837 bp    DNA     circular SYN 01-DEC-2025
DEFINITION  empty vector control for pLJC6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7837)
  AUTHORS   Rossiter NJ, Huggler KS, Adelmann CH, Keys HR, Soens RW, Sabatini 
            DM, Cantor JR
  TITLE     CRISPR screens in physiologic medium reveal conditionally essential 
            genes in human cells.
  JOURNAL   Cell Metab. 2021 Feb 23. pii: S1550-4131(21)00061-9. doi: 
            10.1016/j.cmet.2021.02.005.
  PUBMED    33651980
REFERENCE   2  (bases 1 to 7837)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Metab.
            2021 Feb 23. pii: S1550-4131(21)00061-9. doi: 
            10.1016/j.cmet.2021.02.005."
COMMENT     Sequence Label: pLJC6-EV
FEATURES             Location/Qualifiers
     source          1..7837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             83..263
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(264..490)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(518..538)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(518..536)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(557..573)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(557..573)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(570..592)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    581..597
                     /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(605..611,612..629,630..635))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    650..671
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(788..805)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(959..1547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1039..1058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1718..2509,2510..2578))
                     /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     2341..2360
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2579..2683)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2709..3164)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2846..2867)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3058..3077
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3291..3313
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3305..3322
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3306..3322
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3332..3351
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3332..3350
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      3371..3506
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3546..3667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3557..3576)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3611..3630
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             3739..3972
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(4044..4439)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(4448..4958)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4511..4529)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    5007..5124
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(5172..5237)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(5293..6504)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5437..5456)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             complement(6793..6837)
                     /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    7022..7255
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    join(7748..7837,1..36)
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 gcgcgcttca gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt
       61 caagtccctg ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg
      121 gatctctagt taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc
      181 aagctttatt gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag
      241 atctggtcta accagagaga cccgtttatg tatcgagcta ggcacttaaa tacaatatct
      301 ctgcaatgcg gcaattcagt ggttcgtcca atccatgtca gacccgtctg ttgccttcct
      361 aataaggcac gatcgtacca ccttacttcc accaatcggc atgcacggtg ctttttctct
      421 ccttgtaagg catgttgcta actcatcgtt accatgttgc aagactacaa gagtattgca
      481 taagactaca ttaagcttgc agctccagct tttgttccct ttagtgaggg ttaattgcgc
      541 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
      601 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
      661 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
      721 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
      781 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
      841 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
      901 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      961 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1021 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1081 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1141 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1201 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1261 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1321 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1381 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1441 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1501 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1561 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1621 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1681 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1741 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1801 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     1861 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     1921 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1981 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2041 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2101 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2161 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     2221 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     2281 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     2341 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     2401 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     2461 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     2521 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2581 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2641 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     2701 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2761 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2821 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2881 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2941 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3001 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3061 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3121 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     3181 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     3241 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     3301 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
     3361 tacgtcccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag
     3421 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     3481 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggatagc tagagccaga
     3541 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3601 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3661 acaagttcct ctcactctct gatattcatt tctttgcaag ttataaatac tgaataataa
     3721 gatgacatga actactactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     3781 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     3841 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     3901 ggtctaacca gagagaccca gtacaagcaa aaagcagatc ttgtcttcgt tgggagtgaa
     3961 ttagcccttc cagtcccccc ttttctttta aaaagtggct aagatctaca gctgccttgt
     4021 aagtcattgg tcttaaaggt accttagccc tcccacacat aaccagaggg cagcaattca
     4081 cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca ctatggcttt gatcccagga
     4141 tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc tcaagatgcc cctgttctca
     4201 tttccgatcg cgacgataca agtcaggttg ccagctgccg cagcagcagc agtgcccagc
     4261 accacgagtt ctgcacaagg tcccccagta aaatgatata cattgacacc agtgaagatg
     4321 cggccgtcgc tagagagagc tgcgctggcg acgctgtagt cttcagagat ggggatgctg
     4381 ttgattgtag ccgttgctct ttcaatgagg gtggattctt cttgagacaa aggaaccatg
     4441 gatccccctg gggagagagg tcggtgattc ggtcaacgag ggagccgact gccgacgtgc
     4501 gctccggagg cttgcagaat gcggaacacc gcgcgggcag gaacagggcc cacactaccg
     4561 ccccacaccc cgcctcccgc accgcccctt cccggccgct gctctcggcg cgccctgctg
     4621 agcagccgct attggccaca gcccatcgcg gtcggcgcgc tgccattgct ccctggcgct
     4681 gtccgtctgc gagggtacta gtgagacgtg cggcttccgt ttgtcacgtc cggcacgccg
     4741 cgaaccgcaa ggaaccttcc cgacttaggg gcggagcagg aagcgtcgcc ggggggccca
     4801 caagggtagc ggcgaagatc cgggtgacgc tgcgaacgga cgtgaagaat gtgcgagacc
     4861 cagggtcggc gccgctgcgt ttcccggaac cacgcccaga gcagccgcgt ccctgcgcaa
     4921 acccagggct gccttggaaa aggcgcaacc ccaaccccct cgagccgcgg ccaaagtgga
     4981 tctctgctgt ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat
     5041 tctttagttt gtatgtctgt tgctattatg tctactattc tttcccctgc actgtacccc
     5101 ccaatccccc cttttctttt aaaattgtgg atgaatactg ccatttgtct cgaggtcgag
     5161 aattcttatt acttgtcatc gtcatccttg taatcaatgt catgatcttt ataatcaccg
     5221 tcatggtctt tgtagtcgcc tgcggccgct cgagcttcta agcattaatt aaaccggtag
     5281 cgctagcgct tcgtctaaca aaaaagccaa aaacggccag aatttagcgg acaatttact
     5341 agtctaacac tgaaaattac atattgaccc aaatgattac atttcaaaag gtgcctaaaa
     5401 aacttcacaa aacacactcg ccaaccccga gcgcatagtt caaaaccgga gcttcagcta
     5461 cttaagaaga taggtacata aaaccgacca aagaaactga cgcctcactt atccctcccc
     5521 tcaccagagg tccggcgcct gtcgattcag gagagcctac cctaggcccg aaccctgcgt
     5581 cctgcgacgg agaaaagcct accgcacacc taccggcagg tggccccacc ctgcattata
     5641 agccaacaga acgggtgacg tcacgacacg acgagggcgc gcgctcccaa aggtacgggt
     5701 gcactgccca acggcaccgc cataactgcc gcccccgcaa cagacgacaa accgagttct
     5761 ccagtcagtg acaaacttca cgtcagggtc cccagatggt gccccagccc atctcacccg
     5821 aataagagct ttcccgcatt agcgaaggcc tcaagacctt gggttcttgc cgcccaccat
     5881 gccccccacc ttgtttcaac gacctcacag cccgcctcac aagcgtcttc cattcaagac
     5941 tcgggaacag ccgccatttt gctgcgctcc ccccaacccc cagttcaggg caaccttgct
     6001 cgcggaccca gactacagcc cttggcggtc tctccacacg cttccgtccc accgagcggc
     6061 ccggcggcca cgaaagcccc ggccagccca gcagcccgct actcaccaag tgacgatcac
     6121 agcgatccac aaacaagaac cgcgacccaa atcccggctg cgacggaact agctgtgcca
     6181 cacccggcgc gtccttatat aatcatcggc gttcaccgcc ccacggagat ccctccgcag
     6241 aatcgccgag aagggactac ttttcctcgc ctgttccgct ctctggaaag aaaaccagtg
     6301 ccctagagtc acccaagtcc cgtcctaaaa tgtccttctg ctgatactgg ggttctaagg
     6361 ccgagtctta tgagcagcgg gccgctgtcc tgagcgtccg ggcggaagga tcaggacgct
     6421 cgctgcgccc ttcgtctgac gtggcagcgc tcgccgtgag gaggggggcg cccgcgggag
     6481 gcgccaaaac ccggcgcgga ggccagatcc cgctcgaggc tagtctcgtg atcgataccg
     6541 tcgagatccg ttcactaatc gaatggatct gtctctgtct ctctctccac cttcttcttc
     6601 tattccttcg ggcctgtcgg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg
     6661 tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc
     6721 taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt
     6781 aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc
     6841 ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc
     6901 tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca
     6961 actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc
     7021 caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt
     7081 gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt
     7141 gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga
     7201 cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc
     7261 cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa
     7321 tggttcaatt tttactactt tatatttata taattcactt ctccaattgt ccctcatatc
     7381 tcctcctcca ggtctgaaga tcagctgctg cttgctgtgc ggtggtctta cttttgtttt
     7441 gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc ctttgatgca
     7501 cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat cctgtctgaa
     7561 gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct aacaggccag
     7621 gattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt aatttatatt
     7681 ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt ctcccccgct
     7741 taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc aaaatttttg
     7801 gcgtactcac cagtcgccgc ccctcgcctc ttgccgt
//
