LOCUS       40924_27772        9151 bp DNA     circular SYN 13-MAY-2021
DEFINITION  There is no gene/insert. It is a backbone plasmid that others can 
            use to insert sgRNAs of interest. The cloning site for this BsmBI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9151)
  TITLE     pLenti-sgRNA
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9151)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9151)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     138..157
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     391..408
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    525..546
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        561..591
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    599..615
                     /label=lac operator
                     /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)."
     primer_bind     623..639
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        660..678
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        706..932
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             933..1113
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1160..1285
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1778..2011
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2196..2240
                     /codon_start=1
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
                     /translation="KNEQELLELDKWASL"
     promoter        2418..2658
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        4548..4623
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    4671..4788
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        4857..5068
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             5094..5690
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    5709..6297
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6369..6602
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6680..6814
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6841..6976
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6997..7015)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7025..7041)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7183..7638
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7664..7768
                     /label=AmpR promoter
     CDS             7769..8626
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8800..9151
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
       61 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      121 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      181 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      241 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      301 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      361 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      421 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
      481 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      541 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      601 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
      661 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa
      721 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
      781 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
      841 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
      901 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag
      961 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     1021 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     1081 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     1141 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     1201 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     1261 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     1321 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     1381 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     1441 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     1501 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     1561 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     1621 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     1681 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     1741 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     1801 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     1861 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     1921 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     1981 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     2041 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     2101 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     2161 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     2221 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     2281 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     2341 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     2401 ccccgagggg acccagagag ggcctatttc ccatgattcc ttcatatttg catatacgat
     2461 acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag atattagtac
     2521 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt
     2581 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat
     2641 atatcttgtg gaaaggacga aacaccggag acggttgtaa atgagcacac aaaatacaca
     2701 tgctaaaata ttatattcta tgacctttat aaaatcaacc aaaatcttct ttttaataac
     2761 tttagtatca ataattagaa tttttatgtt cctttttgca aacttttaat aaaaatgagc
     2821 aaaataaaaa aacgctagtt ttagtaactc gcgttgtttt cttcaccttt aataatagct
     2881 actccaccac ttgttcctaa gcggtcagct cctgcttcaa tcattttttg agcatcttca
     2941 aatgttctaa ctccaccagc tgctttaact aaagcattgt ctttaacaac tgacttcatt
     3001 agtttaacat cttcaaatgt tgcacctgat tttgaaaatc ctgttgatgt tttaacaaat
     3061 tctaatccag cttcaacagc tatttcacaa gctttcatga tttcttcttt tgttaataaa
     3121 caattttcca taatacattt aacaacatgt gatccagctg ctttttttac agctttcatg
     3181 tcttctaaaa ctaattcata atttttgtct tttaatgcac caatatttaa taccatatca
     3241 atttctgttg caccatcttt aattgcttca gaaacttcga atgcttttgt agctgttgtg
     3301 catgcaccta gaggaaaacc tacaacattt gttattccta catttgtgcc ttttaataat
     3361 tctttacaat agcttgttca atatgaatta acacaaactg ttgcaaaatc aaattcaatt
     3421 gcttcatcac ataattgttt aatttcagct ttcgtagcat cttgttttaa taatgtgtga
     3481 tctatatatt tgtttagttt cattttttct cctatatatt catttttaat tttaattctt
     3541 taataatttc gtctacttta actttagcgt tttgaacaga ttcaccaaca cctataaaat
     3601 aaatttttag tttaggttca gttccacttg ggcgaacagc aaatcatgac ttatcttcta
     3661 aataaaattt tagtaagtct tgtcctggca tattatacat tccatcgatg tagtcttcaa
     3721 cattaacaac tttaagtcca gcaatttgag ttaagggtgt tgctctcaat gatttcatta
     3781 atggttcaat ttttaatttc ttttcttctg gtttaaaatt caagtttaaa gtgaaagtgt
     3841 aatatgcacc catttcttta aataaatctt ctaaatagtc tactaatgtt ttattttgtt
     3901 ttttataaaa tcaagcagcc tctgctatta atatagaagc ttgtattcca tctttatctc
     3961 tagctgagtc atcaattaca tatccataac tttcttcata agcaaaaaca aaatttaatc
     4021 cgttatcttc ttctttagca atttctctac ccattcattt aaatccagtt aaagttttta
     4081 caatattaac tccatatttt tcatgagcga ttctatcacc caaatcactt gttacaaaac
     4141 ttgaatatag agccggattt tttggaatgc tatttaagcg ttttagattt gataattttc
     4201 aatcaattaa aattggtcct gtttgatttc catctaatct tacaaaatga ccatcatgtt
     4261 ttattgccat tccaaatctg tcagcatctg ggtcattcat aataataata tctgcatcat
     4321 gtttaatacc atattcaagc ggtatttttc atgcaggatc aaattctgga tttggattta
     4381 caacattttt aaatgtttca tcttcaaatg catgctcttc aacctcaata acgttatatc
     4441 ctgattcacg taatattttt ggggtaaatt tagttcctgt tccattaact gcgctaaaaa
     4501 taatttttaa atctttttta gcttcttgct cttttttgta cgtctctgtt ttagagctag
     4561 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
     4621 tgcttttttg aattctagat cttgagacaa atggcagtat tcatccacaa ttttaaaaga
     4681 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     4741 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     4801 agggacagca gagatccact ttggcgccgg ctcgagtggc tccggtgccc gtcagtgggc
     4861 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg
     4921 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     4981 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     5041 tcgcaacggg tttgccgcca gaacacaggt gtcgtgacgc gggatccgcc accatgaccg
     5101 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     5161 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     5221 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     5281 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     5341 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     5401 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     5461 ggttcctggc caccgtcgga gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     5521 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     5581 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     5641 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaacgcgtt
     5701 aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     5761 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     5821 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     5881 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     5941 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     6001 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     6061 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     6121 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     6181 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     6241 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc
     6301 gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     6361 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     6421 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     6481 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6541 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6601 cagtacgtat agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     6661 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     6721 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6781 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     6841 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6901 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     6961 ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc
     7021 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     7081 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     7141 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     7201 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     7261 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     7321 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     7381 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     7441 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     7501 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     7561 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     7621 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     7681 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     7741 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     7801 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     7861 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7921 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     7981 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     8041 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     8101 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     8161 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     8221 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     8281 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     8341 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     8401 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     8461 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     8521 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     8581 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     8641 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     8701 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     8761 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     8821 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     8881 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     8941 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     9001 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     9061 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     9121 ggggggttcg tgcacacagc ccagcttgga g
//
