LOCUS       Exported                6633 bp ds-DNA     circular SYN 03-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6633)
  AUTHORS   Ryan Lister
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Oct 31, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6633
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    6..230
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6..27
                     /label=OC-310_pLenti_1_dCas9_5.F
     primer_bind     complement(204..230)
                     /label=OC-311_pLenti_1_dCas9_5.R
     primer_bind     complement(210..253)
                     /label=TOL827_pGHpolyA_R
     primer_bind     211..254
                     /label=TOL825_pEF1a-GFP_R
     CDS             complement(240..956)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     240..260
                     /label=TOL195_GFP_PUF_polyA_R
     primer_bind     complement(666..684)
                     /label=TOL102_GFP_backbone_F
     primer_bind     682..696
                     /label=TOL103_GFP_backbone_R
     primer_bind     complement(939..956)
                     /label=TOL194_T2A_GFP_F
     primer_bind     complement(949..988)
                     /label=TOL837_GFP_bGHpolyA_F
     primer_bind     949..987
                     /label=TOL840_pSV40_R
     promoter        complement(981..1191)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      987..1122
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1170..1224)
                     /label=TOL839_pSV40_F
     primer_bind     1183..1225
                     /label=TOL847_pEF1a_F
     primer_bind     complement(1212..1229)
                     /label=TOL847_pEF1a_F
     primer_bind     1213..1229
                     /label=TOL839_pSV40_F
     primer_bind     complement(1291..1312)
                     /label=TOL319_GCN4_pEF1a_R
     primer_bind     1291..1312
                     /label=TOL318_GCN4_pEF1a_F
     primer_bind     1331..1356
                     /label=OL-D206
     promoter        1406..2663
                     /label=EF1a
     primer_bind     complement(1478..1501)
                     /label=OL-D243
     primer_bind     1846..1867
                     /label=OL -D154
     primer_bind     2458..2479
                     /label=OL- D155
     primer_bind     2524..2545
                     /label=TOL417_GCN4_seq_F
     primer_bind     complement(2655..2674)
                     /label=TOL848_pEF1a_R
     misc_feature    2670..2675
                     
     primer_bind     complement(2673..2702)
                     /label=TOL401_3xTy1_GCN4_ R
     CDS             2676..2678
                     /codon_start=1
                     /label=ATG
                     /translation="M"
     CDS             2682..2702
                     /codon_start=1
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2703..2744
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     CDS             2745..2786
                     /codon_start=1
                     /product="Escherichia coli OmpF Linker and mouse Langerin 
                     fusion sequence tag (Park et al., 2008)"
                     /label=OLLAS tag
                     /translation="SGFANELGPRLMGK"
     primer_bind     2786..2804
                     /label=TOL779_SnoopCatcher_R
     CDS             2787..3122
                     /codon_start=1
                     /label=SnoopCatcher
                     /translation="KPLRGAVFSLQKQHPDYPDIYGAIDQNGTYQNVRTGEDGKLTFKN
                     LSDGKYRLFENSEPAGYKPVQNKPIVAFQIVNGEVRDVTSIVPQDIPATYEFTNGKHYI
                     TNEPIPPK"
     primer_bind     complement(3098..3142)
                     /label=TOL781_SnoopCatcher_F2
     primer_bind     3103..3139
                     /label=TOL780_linker_p65HSF1_F2
     CDS             3123..3176
                     /codon_start=1
                     /label=Linker
                     /translation="SAGGGGSGGGGSGGGGSG"
     primer_bind     3123..3160
                     /label=TOL35
     primer_bind     3123..3142
                     /label=TOL774_linker_p65HSF1_F
     primer_bind     3145..3163
                     /label=TOL418_GB1linker_seq_F
     primer_bind     3160..3208
                     /label=TOL317_p65HSF1_GCN4_F2
     primer_bind     complement(3160..3205)
                     /label=TOL316_GCN4_p65HSF1_R2
     primer_bind     complement(3164..3185)
                     /label=TOL-29
     primer_bind     complement(3167..3185)
                     /label=TOL405
     CDS             3186..3728
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     primer_bind     3186..3208
                     /label=TOL845_p65HSF1_F
     CDS             3753..3824
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSL"
     primer_bind     complement(3806..3824)
                     /label=TOL409_p65HSF1_codon_R
     misc_feature    3825..4124
                     /label=p65HSF1 codon change
     primer_bind     3923..3942
                     /label=TOL465_p65HSF1_qPCR1_F
     primer_bind     complement(4045..4064)
                     /label=TOL466_p65HSF1_qPCR1_R
     primer_bind     complement(4128..4152)
                     /label=TOL614_UPS_PolyT_R
     misc_feature    complement(4128..4151)
                     /label=UPS3
     polyA_signal    4152..4390
                     /label=SV40 polyA
     primer_bind     4152..4191
                     /label=TOL400_UPS_polyA_F
     primer_bind     complement(4212..4236)
                     /label=TOL419_SV40_seq_R
     rep_origin      4869..5488
                     /label=pBR322 Orgin
     CDS             complement(5643..6503)
                     /codon_start=1
                     /label=Ampicillin
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6545..6573)
                     /label=AMP promoter
     primer_bind     join(6616..6633,1..25)
                     /label=TOL838_GFP_bGHpolyA_R
     primer_bind     complement(join(6618..6633,1..25))
                     /label=TOL846_p65HSF1_R
ORIGIN
        1 gacgtccata gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc
       61 cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga
      121 tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga
      181 aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcaactgccc
      241 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
      301 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg
      361 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
      421 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc
      481 ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
      541 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
      601 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
      661 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
      721 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg
      781 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
      841 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
      901 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg
      961 gccgtacggg tggcgcgcgt agcctaggcc tccaaaaaag cctcctcact acttctggaa
     1021 tagctcagag gcagaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     1081 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     1141 gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg cctgaattca
     1201 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     1261 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     1321 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     1381 ctgcttaggg ttaggcgttt tgcgtgcaaa gatggataaa gttttaaaca gagaggaatc
     1441 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
     1501 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     1561 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     1621 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     1681 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
     1741 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc
     1801 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt
     1861 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc
     1921 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
     1981 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
     2041 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
     2101 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
     2161 tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt
     2221 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
     2281 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag
     2341 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
     2401 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt
     2461 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
     2521 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
     2581 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
     2641 ttcttccatt tcaggtgtcg tgacgtacgg ccaccatggg tcctaagaaa aagaggaagg
     2701 tgtccggctt tgcgaacgaa cttggacctc gattgatggg caagagcggc ttcgccaacg
     2761 agctgggccc cagactgatg ggcaaaaagc cgctgcgtgg tgccgtgttt agcctgcaga
     2821 aacagcatcc cgactatccc gatatctatg gcgcgattga tcagaatggg acctatcaaa
     2881 atgtgcgtac cggcgaagat ggtaaactga cctttaagaa tctgagcgat ggcaaatatc
     2941 gcctgtttga aaatagcgaa cccgctggct ataaaccggt gcagaataag ccgattgtgg
     3001 cgtttcagat tgtgaatggc gaagtgcgtg atgtgaccag cattgtgccg caggatattc
     3061 cggctacata tgaatttacc aacggtaaac attatatcac caatgaaccg ataccgccga
     3121 aaagcgctgg agggggtggc tccggcggag gaggctctgg tggcggaggg agtggagcgg
     3181 ccgctccttc agggcagatc agcaaccagg ccctggctct ggcccctagc tccgctccag
     3241 tgctggccca gactatggtg ccctctagtg ctatggtgcc tctggcccag ccacctgctc
     3301 cagcccctgt gctgacccca ggaccacccc agtcactgag cgctccagtg cccaagtcta
     3361 cacaggccgg cgaggggact ctgagtgaag ctctgctgca cctgcagttc gacgctgatg
     3421 aggacctggg agctctgctg gggaacagca ccgatcccgg agtgttcaca gatctggcct
     3481 ccgtggacaa ctctgagttt cagcagctgc tgaatcaggg cgtgtccatg tctcatagta
     3541 cagccgaacc aatgctgatg gagtaccccg aagccattac ccggctggtg accggcagcc
     3601 agcggccccc cgaccccgct ccaactcccc tgggaaccag cggcctgcct aatgggctgt
     3661 ccggagatga agacttctca agcatcgctg atatggactt tagtgccctg ctgtcacaga
     3721 tttcctctag tgggcaggga ggaggtggaa gcggcttcag cgtggacacc agtgccctgc
     3781 tggacctgtt cagcccctcg gtgaccgtgc ccgacatgag cctgcctgat cttgacagtt
     3841 cattggcgag cattcaagag ttgttgtccc cgcaggaacc gccccgaccg ccggaagcgg
     3901 agaacagcag ccctgactca ggaaagcagc tggtccacta cacagcccaa ccactgtttt
     3961 tgcttgatcc aggtagtgtt gacaccggct caaatgactt gcctgtcctc tttgaactcg
     4021 gtgaggggtc ttacttcagc gagggagacg gtttcgcaga agacccaact atttccctcc
     4081 tcactggcag cgagccccct aaagcgaaag atcccacggt atcctgacgt tgagtctctg
     4141 tttgtggtga ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4201 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4261 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4321 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4381 atgcgtcgag tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4441 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4501 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4561 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4621 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4681 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4741 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4801 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4861 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4921 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4981 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5041 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5101 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5161 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5221 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5281 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5341 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5401 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5461 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5521 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5581 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5641 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5701 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5761 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5821 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5881 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5941 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6001 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6061 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6121 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6181 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6241 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6301 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6361 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6421 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6481 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6541 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6601 gttccgcgca catttccccg aaaagtgcca cct
//
