LOCUS       Exported                9342 bp ds-DNA     circular SYN 23-JAN-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pLKO.1C
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9342)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jan 23, 2020 from SnapGene Viewer 4.3.9
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9342
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             169..768
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /note="color: #ccffcc"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     LTR             896..1129
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
                     /note="color: #ffe4c4"
     polyA_signal    1201..1322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      1362..1497
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     promoter        complement(1518..1536)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(1546..1562)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     rep_origin      1704..2159
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        2185..2289
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             2290..3150
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 2290 .. 2358 / #ccffcc / signal sequence
                        2: 2359 .. 3150 / #ccffcc
                     Cleavage site after base 2358"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3321..3909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     protein_bind    4197..4218
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        4233..4263
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 4233 .. 4238 / #ffffff / -35
                        2: 4239 .. 4256 / #ffffff
                        3: 4257 .. 4263 / #ffffff / -10"
     protein_bind    4271..4287
                     /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)."
                     /note="color: #31849b"
     primer_bind     4295..4311
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        4332..4350
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     enhancer        4451..4830
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        4831..5034
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     LTR             5049..5229
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     misc_feature    5273..5398
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /note="color: #ffe4c4"
     misc_feature    5891..6124
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
                     /note="color: #ffe4c4"
     promoter        6651..6891
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    8813..8930
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
                     /note="color: #ffe4c4"
     promoter        join(8979..9342,1..147)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
                     /note="color: #ffffff; direction: RIGHT"
ORIGIN
        1 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
       61 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      121 gaccgaatca ccgacctctc tccccagggg gatccaccgg agcttaccat gaccgagtac
      181 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
      241 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
      301 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
      361 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
      421 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
      481 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
      541 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
      601 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
      661 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
      721 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
      781 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcggtacc tttaagacca
      841 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
      901 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
      961 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     1021 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     1081 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     1141 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     1201 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     1261 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     1321 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
     1381 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     1441 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     1501 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     1561 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     1621 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     1681 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1741 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     1801 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     1861 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     1921 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     1981 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     2041 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     2101 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     2161 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2221 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2281 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2341 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2401 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2461 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2521 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2581 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2641 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2701 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2761 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2821 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2881 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2941 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3001 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3061 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3121 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3181 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3241 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3301 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3361 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3421 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3481 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3541 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3601 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3661 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3721 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3781 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3841 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3901 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3961 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4021 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4081 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4141 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     4201 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     4261 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     4321 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttggcca
     4381 ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg tccaacatta
     4441 ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     4501 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     4561 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     4621 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     4681 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     4741 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     4801 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     4861 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     4921 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     4981 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
     5041 gtgaaccggg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     5101 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     5161 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     5221 tctctagcag tggcgcccga acagggacct gaaagcgaaa gggaaaccag agctctctcg
     5281 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     5341 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     5401 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     5461 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     5521 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     5581 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     5641 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     5701 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     5761 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     5821 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     5881 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     5941 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     6001 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     6061 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     6121 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     6181 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     6241 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     6301 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     6361 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     6421 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     6481 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     6541 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     6601 ctcgacggta tcgatcacga gactagcctc gagcggccgc ccccttcacc gagggcctat
     6661 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
     6721 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     6781 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     6841 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
     6901 gtgtgttgta aatgagcaca caaaatacac atgctaaaat attatattct atgaccttta
     6961 taaaatcaac caaaatcttc tttttaataa ctttagtatc aataattaga atttttatgt
     7021 tcctttttgc aaacttttaa taaaaatgag caaaataaaa aaacgctagt tttagtaact
     7081 cgcgttgttt tcttcacctt taataatagc tactccacca cttgttccta agcggtcagc
     7141 tcctgcttca atcatttttt gagcatcttc aaatgttcta actccaccag ctgctttaac
     7201 taaagcattg tctttaacaa ctgacttcat tagtttaaca tcttcaaatg ttgcacctga
     7261 ttttgaaaat cctgttgatg ttttaacaaa ttctaatcca gcttcaacag ctatttcaca
     7321 agctttcatg atttcttctt ttgttaataa acaattttcc ataatacatt taacaacatg
     7381 tgatccagct gcttttttta cagctttcat gtcttctaaa actaattcat aatttttgtc
     7441 ttttaatgca ccaatattta ataccatatc aatttctgtt gcaccatctt taattgcttc
     7501 agaaacttcg aatgcttttg tagctgttgt gcatgcacct agaggaaaac ctacaacatt
     7561 tgttattcct acatttgtgc cttttaataa ttctttacaa tagcttgttc aatatgaatt
     7621 aacacaaact gttgcaaaat caaattcaat tgcttcatca cataattgtt taatttcagc
     7681 tttcgtagca tcttgtttta ataatgtgtg atctatatat ttgtttagtt tcattttttc
     7741 tcctatatat tcatttttaa ttttaattct ttaataattt cgtctacttt aactttagcg
     7801 ttttgaacag attcaccaac acctataaaa taaattttta gtttaggttc agttccactt
     7861 gggcgaacag caaatcatga cttatcttct aaataaaatt ttagtaagtc ttgtcctggc
     7921 atattataca ttccatcgat gtagtcttca acattaacaa ctttaagtcc agcaatttga
     7981 gttaagggtg ttgctctcaa tgatttcatt aatggttcaa tttttaattt cttttcttct
     8041 ggtttaaaat tcaagtttaa agtgaaagtg taatatgcac ccatttcttt aaataaatct
     8101 tctaaatagt ctactaatgt tttattttgt tttttataaa atcaagcagc ctctgctatt
     8161 aatatagaag cttgtattcc atctttatct ctagctgagt catcaattac atatccataa
     8221 ctttcttcat aagcaaaaac aaaatttaat ccgttatctt cttctttagc aatttctcta
     8281 cccattcatt taaatccagt taaagttttt acaatattaa ctccatattt ttcatgagcg
     8341 attctatcac ccaaatcact tgttacaaaa cttgaatata gagccggatt ttttggaatg
     8401 ctatttaagc gttttagatt tgataatttt caatcaatta aaattggtcc tgtttgattt
     8461 ccatctaatc ttacaaaatg accatcatgt tttattgcca ttccaaatct gtcagcatct
     8521 gggtcattca taataataat atctgcatca tgtttaatac catattcaag cggtattttt
     8581 catgcaggat caaattctgg atttggattt acaacatttt taaatgtttc atcttcaaat
     8641 gcatgctctt caacctcaat aacgttatat cctgattcac gtaatatttt tggggtaaat
     8701 ttagttcctg ttccattaac tgcgctaaaa ataattttta aatctttttt agcttcttgc
     8761 tcttttttgt agaattctcg acctcgagac aaatggcagt attcatccac aattttaaaa
     8821 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     8881 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     8941 acagggacag cagagatcca ctttggccgc ggctcgaggg ggttggggtt gcgccttttc
     9001 caaggcagcc ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc
     9061 agcggcgccg accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc
     9121 ttcgccgcta cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa
     9181 ggttccttgc ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt
     9241 accctcgcag acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg
     9301 gccaatagcg gctgctcagc agggcgcgcc gagagcagcg gc
//
