LOCUS       Exported                9407 bp ds-DNA     circular SYN 28-JAN-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pLKO.1C-YFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9407)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 28, 2020 from SnapGene Viewer 4.3.9
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9407
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      169..178
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             175..894
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 175 .. 177 / #e1ff2f
                        2: 178 .. 180 / #e1ff2f / 1a
                        3: 181 .. 894 / #e1ff2f"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     LTR             961..1194
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
                     /note="color: #ffe4c4"
     polyA_signal    1266..1387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      1427..1562
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     promoter        complement(1583..1601)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(1611..1627)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     rep_origin      1769..2224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        2250..2354
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             2355..3215
                     /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: 2355 .. 2423 / #ccffcc / signal sequence
                        2: 2424 .. 3215 / #ccffcc
                     Cleavage site after base 2423"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3386..3974
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     protein_bind    4262..4283
                     /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        4298..4328
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 4298 .. 4303 / #ffffff / -35
                        2: 4304 .. 4321 / #ffffff
                        3: 4322 .. 4328 / #ffffff / -10"
     protein_bind    4336..4352
                     /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     4360..4376
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        4397..4415
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     enhancer        4516..4895
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        4896..5099
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     LTR             5114..5294
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     misc_feature    5338..5463
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /note="color: #ffe4c4"
     misc_feature    5956..6189
                     /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        6716..6956
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    8878..8995
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
                     /note="color: #ffe4c4"
ORIGIN
        1 gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt agtgtgggcc
       61 ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac gtcggcagtc
      121 ggctccctcg ttgaccgaat caccgacctc tctccccagg gggatctcgc caccatggtg
      181 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      241 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      301 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      361 accaccttcg gctacggcct gaagtgcttc gcccgctacc ccgaccacat gaagcagcac
      421 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      481 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      541 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
      601 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
      661 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
      721 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
      781 agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
      841 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaactttaa
      901 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
      961 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     1021 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     1081 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     1141 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     1201 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     1261 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1321 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1381 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
     1441 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     1501 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     1561 cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     1621 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     1681 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     1741 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     1801 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1861 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     1921 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     1981 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2041 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2101 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2161 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     2221 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2281 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2341 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2401 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2461 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2521 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2581 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2641 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2701 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2761 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2821 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2881 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     2941 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3001 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3061 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3121 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3181 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3241 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3301 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3361 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3421 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3481 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3541 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3601 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3661 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3721 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3781 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3841 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3901 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3961 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4021 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4081 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4141 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4201 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     4261 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     4321 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     4381 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt
     4441 ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa
     4501 cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt
     4561 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4621 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4681 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     4741 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     4801 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4861 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     4921 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     4981 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     5041 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     5101 gtttagtgaa ccggggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     5161 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     5221 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     5281 gaaaatctct agcagtggcg cccgaacagg gacctgaaag cgaaagggaa accagagctc
     5341 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
     5401 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     5461 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     5521 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     5581 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     5641 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     5701 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     5761 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
     5821 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     5881 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     5941 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     6001 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     6061 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     6121 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     6181 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     6241 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     6301 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     6361 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     6421 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     6481 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     6541 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     6601 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     6661 cggatctcga cggtatcgat cacgagacta gcctcgagcg gccgccccct tcaccgaggg
     6721 cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta gagagataat
     6781 tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta
     6841 ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct
     6901 taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa
     6961 caccggtgtg ttgtaaatga gcacacaaaa tacacatgct aaaatattat attctatgac
     7021 ctttataaaa tcaaccaaaa tcttcttttt aataacttta gtatcaataa ttagaatttt
     7081 tatgttcctt tttgcaaact tttaataaaa atgagcaaaa taaaaaaacg ctagttttag
     7141 taactcgcgt tgttttcttc acctttaata atagctactc caccacttgt tcctaagcgg
     7201 tcagctcctg cttcaatcat tttttgagca tcttcaaatg ttctaactcc accagctgct
     7261 ttaactaaag cattgtcttt aacaactgac ttcattagtt taacatcttc aaatgttgca
     7321 cctgattttg aaaatcctgt tgatgtttta acaaattcta atccagcttc aacagctatt
     7381 tcacaagctt tcatgatttc ttcttttgtt aataaacaat tttccataat acatttaaca
     7441 acatgtgatc cagctgcttt ttttacagct ttcatgtctt ctaaaactaa ttcataattt
     7501 ttgtctttta atgcaccaat atttaatacc atatcaattt ctgttgcacc atctttaatt
     7561 gcttcagaaa cttcgaatgc ttttgtagct gttgtgcatg cacctagagg aaaacctaca
     7621 acatttgtta ttcctacatt tgtgcctttt aataattctt tacaatagct tgttcaatat
     7681 gaattaacac aaactgttgc aaaatcaaat tcaattgctt catcacataa ttgtttaatt
     7741 tcagctttcg tagcatcttg ttttaataat gtgtgatcta tatatttgtt tagtttcatt
     7801 ttttctccta tatattcatt tttaatttta attctttaat aatttcgtct actttaactt
     7861 tagcgttttg aacagattca ccaacaccta taaaataaat ttttagttta ggttcagttc
     7921 cacttgggcg aacagcaaat catgacttat cttctaaata aaattttagt aagtcttgtc
     7981 ctggcatatt atacattcca tcgatgtagt cttcaacatt aacaacttta agtccagcaa
     8041 tttgagttaa gggtgttgct ctcaatgatt tcattaatgg ttcaattttt aatttctttt
     8101 cttctggttt aaaattcaag tttaaagtga aagtgtaata tgcacccatt tctttaaata
     8161 aatcttctaa atagtctact aatgttttat tttgtttttt ataaaatcaa gcagcctctg
     8221 ctattaatat agaagcttgt attccatctt tatctctagc tgagtcatca attacatatc
     8281 cataactttc ttcataagca aaaacaaaat ttaatccgtt atcttcttct ttagcaattt
     8341 ctctacccat tcatttaaat ccagttaaag tttttacaat attaactcca tatttttcat
     8401 gagcgattct atcacccaaa tcacttgtta caaaacttga atatagagcc ggattttttg
     8461 gaatgctatt taagcgtttt agatttgata attttcaatc aattaaaatt ggtcctgttt
     8521 gatttccatc taatcttaca aaatgaccat catgttttat tgccattcca aatctgtcag
     8581 catctgggtc attcataata ataatatctg catcatgttt aataccatat tcaagcggta
     8641 tttttcatgc aggatcaaat tctggatttg gatttacaac atttttaaat gtttcatctt
     8701 caaatgcatg ctcttcaacc tcaataacgt tatatcctga ttcacgtaat atttttgggg
     8761 taaatttagt tcctgttcca ttaactgcgc taaaaataat ttttaaatct tttttagctt
     8821 cttgctcttt tttgtagaat tctcgacctc gagacaaatg gcagtattca tccacaattt
     8881 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     8941 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     9001 ttattacagg gacagcagag atccactttg gccgcggctc gagggggttg gggttgcgcc
     9061 ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga
     9121 aacgcagcgg cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc
     9181 ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc
     9241 gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca
     9301 ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg
     9361 ctgtggccaa tagcggctgc tcagcagggc gcgccgagag cagcggc
//
