LOCUS       V000729                 8102 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V000729
VERSION     V000729
KEYWORDS    pLKO.3G
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8102)
  TITLE     pLKO variants
REFERENCE   2  (bases 1 to 8102)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8102)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    89..110
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        125..155
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    163..179
                     /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     187..203
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        224..242
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        270..496
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             497..677
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    724..849
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1342..1575
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1760..1804
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             1953..1994
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     promoter        2321..2561
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    2661..2778
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2827..3337
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3359..4075
                     /label="EGFP"
                     /note="enhanced GFP"
     misc_feature    4097..4685
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(4688..4704)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     LTR             4890..5123
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5195..5329
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5356..5491
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(5512..5530)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5540..5556)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      5698..6153
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6179..6283
                     /label="AmpR promoter"
     CDS             6284..7141
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      7315..7903
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     8057..8074
                     /label="L4440"
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
       61 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
      121 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
      181 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
      241 gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg
      301 caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg
      361 ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct
      421 gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct
      481 agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
      541 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
      601 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
      661 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
      721 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
      781 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
      841 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
      901 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
      961 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     1021 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     1081 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     1141 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     1201 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     1261 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     1321 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     1381 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     1441 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     1501 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     1561 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     1621 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     1681 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     1741 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     1801 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     1861 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     1921 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     1981 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     2041 agtgaacgga tctcgacggt atcgatcacg agactagcct cgaccacgac gcaatggaga
     2101 aagacggaga gcgccaacgg cgtccatctc gaaggagtcg ccagcgataa ccggagtagt
     2161 tgaaatggta ataagacgac caatctgacc agcaaggaag ccaagatggg aaaggtcatg
     2221 cggcatacgc tcggcgccag tttgaatatt agacataatt tatcctcaag taagggggta
     2281 cacgcgtcag tcgagtcctg tcgacaaggt cgggcaggaa gagggcctat ttcccatgat
     2341 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac
     2401 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
     2461 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
     2521 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg gtccgcaggt
     2581 atgcttgcgt gaattccacc tcgagctact taattaaacc agttttcgac ctcgagacaa
     2641 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     2701 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     2761 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact ttggccgcgg
     2821 ctcgaggggg ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg
     2881 ctgctctggg cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt
     2941 cacgtccgtt cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac
     3001 gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg
     3061 acaaacggaa gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat
     3121 ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga
     3181 gagcagcggc cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct
     3241 gttcctgccc gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg
     3301 ctccctcgtt gaccgaatca ccgacctctc tccccagggg gatccaccgg tcgccaccat
     3361 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     3421 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     3481 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     3541 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     3601 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     3661 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3721 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3781 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     3841 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     3901 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     3961 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     4021 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaga
     4081 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4141 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4201 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     4261 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     4321 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4381 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     4441 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     4501 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     4561 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4621 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     4681 cccgcatcga taccgtcgac ctcgagacct agaaaaacat ggagcaatca caagtagcaa
     4741 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
     4801 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     4861 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     4921 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     4981 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     5041 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     5101 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     5161 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     5221 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5281 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     5341 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5401 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     5461 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
     5521 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     5581 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     5641 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     5701 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5761 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5821 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5881 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5941 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6001 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     6061 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6121 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     6181 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6241 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6301 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6361 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6421 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6481 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6541 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6601 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6661 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6721 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6781 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6841 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6901 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6961 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7021 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7081 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7141 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7201 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7261 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7321 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7381 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7441 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7501 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7561 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7621 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7681 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7741 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7801 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7861 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7921 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7981 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     8041 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     8101 ac
//
