LOCUS       Exported                8469 bp ds-DNA     circular SYN 25-OCT-2019
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1-kozak-AtCSC1-nostop-pp-EGFP-flag fusion
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8469)
  AUTHORS   Patapoutian Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Oct 25, 2019 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8469
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-Forward
     primer_bind     863..882
                     /label=T7
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     925..962
                     /label=AtCSC1-NotI-Kozak_F
     primer_bind     935..961
                     /label=AtCSC1_Kozak_XbaI_Fwd
     CDS             941..3253
                     /codon_start=1
                     /label=AtCSC1
                     /translation="MATLQDIGVSAGINILSAFVFFIIFAVLRLQPFNDRVYFSKWYLK
                     GLRSSPARGGAFAQRFVNLDFRSYMKFLNWMPEALKMPEPELIDHAGLDSVVYLRIYWL
                     GLKIFTPIAVLAWAVLVPVNWTNNTLEMAKQLRNVTSSDIDKLSVSNIPEYSMRFWTHI
                     VMAYAFTIWTCYVLMKEYETIANMRLQFVASEARRPDQFTVLVRNVPPDADESVSELVE
                     HFFLVNHPDHYLTHQVVCNANKLADLVKKKKKLQNWLDYYQLKYARNNSQRIMVKLGFL
                     GLWGQKVDAIEHYIAEIDKISKEISKEREEVVNDPKAIMPAAFVSFKTRWAAAVCAQTQ
                     QTRNPTQWLTEWAPEPRDVFWSNLAIPYVSLTVRRLIMHVAFFFLTFFFIVPIAFVQSL
                     ATIEGIVKAAPFLKFIVDDKFMKSVIQGFLPGIALKLFLAFLPSILMIMSKFEGFTSIS
                     SLERRAAFRYYIFNLVNVFLASVIAGAAFEQLNSFLNQSANQIPKTIGVAIPMKATFFI
                     TYIMVDGWAGVAGEILMLKPLIMFHLKNAFLVKTDKDREEAMDPGSIGFNTGEPRIQLY
                     FLLGLVYAPVTPMLLPFILVFFALAYIVYRHQIINVYNQEYESAAAFWPDVHGRVIAAL
                     VISQLLLMGLLGTKHAALAAPFLIALPVLTIGFHHFCKGRYEPAFIRYPLQEAMMKDTL
                     ETAREPNLNLKGYLQNAYVHPVFKGDEDDYDIDDKLGKFEDEAIIVPTKRQSRRNTPAP
                     SIISGDDSPSLPFSGKLV"
     primer_bind     941..961
                     /label=AtCSC1_XbaI_Fwd
     primer_bind     941..961
                     /label=AtCSC1-XbaI_F
     primer_bind     1504..1516
                     /label=EGFP-C
     primer_bind     1595..1615
                     /label=AtCSC1_Seq_654_F
     primer_bind     1796..1814
                     /label=AtCSC1-seq_F
     primer_bind     2704..2722
                     /label=AtCSC1_Seq_1763_F
     primer_bind     complement(3234..3262)
                     /label=AtCSC1-NoStop-AgeI_R
     primer_bind     complement(3235..3262)
                     /label=AtCSC1_AgeI_Rev
     primer_bind     complement(3235..3262)
                     /label=AtCSC1-AgeI_R
     CDS             3266..3289
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     misc_feature    3290..4002
                     /label=EGFP
     primer_bind     3941..3961
                     /label=EGFP-C
     CDS             4010..4033
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4063..4080)
                     /label=BGHR
     polyA_signal    4069..4293
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4236..4255
                     /label=BGH 3' F Seq
     rep_origin      4339..4767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4781..5110
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4961..5096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5023..5042
                     /label=SV40-promoter
     CDS             5177..5971
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5502..5519)
                     /label=Neo-R
     primer_bind     5841..5860
                     /label=Neo-F
     polyA_signal    6145..6266
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6182..6201)
                     /label=SV40pA-R
     primer_bind     complement(6315..6331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6315..6331)
                     /label=M13R
     promoter        complement(6363..6393)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(6717..7302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7473..8333)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8096..8115)
                     /label=AmpR F Seq
     primer_bind     8096..8115
                     /label=AmpR R Seq
     promoter        complement(8334..8438)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccgccacc atggctaccc tgcaagatat
      961 cggcgtgagc gctggaatca acatcctctc cgcctttgtg ttcttcatca tcttcgctgt
     1021 gctgagactg cagcccttta acgacagggt ctacttcagc aagtggtacc tgaagggact
     1081 caggtccagc cctgctaggg gaggagcctt cgctcagaga ttcgtgaacc tcgacttcag
     1141 gagctacatg aagttcctga actggatgcc tgaggctctc aaaatgcccg agcccgagct
     1201 gattgatcac gccggcctgg atagcgtcgt ctatctgagg atctactggc tcggcctgaa
     1261 gatctttaca cctatcgccg tgctggcctg ggctgtcctg gtgcccgtga attggaccaa
     1321 caacacactg gagatggcca aacagctgag gaatgtgacc tcctccgaca tcgacaaact
     1381 gagcgtctcc aacatccccg agtatagcat gaggttctgg acccacatcg tcatggccta
     1441 cgcctttacc atctggacct gctacgtcct gatgaaggag tacgagacca tcgccaacat
     1501 gaggctgcag ttcgtggcca gcgaagctag aaggcccgac cagtttaccg tgctggtgag
     1561 gaacgtgccc cctgacgctg acgaatccgt gagcgagctc gtcgagcatt tcttcctggt
     1621 gaaccatccc gaccactacc tgacccacca agtggtgtgc aacgccaaca aactcgccga
     1681 cctggtcaaa aaaaaaaaga agctccagaa ctggctggac tactaccagc tcaagtacgc
     1741 caggaacaat agccagagga ttatggtcaa gctcggcttt ctgggcctct ggggccagaa
     1801 ggtcgatgcc attgagcact acatcgccga gattgacaaa atctccaagg agatctccaa
     1861 agagagggag gaagtggtga acgaccccaa agccatcatg cctgccgcct tcgtgagctt
     1921 taagaccagg tgggctgccg ccgtgtgcgc tcagacccaa cagaccagaa accccaccca
     1981 gtggctcacc gaatgggctc ctgaacccag ggacgtgttc tggtccaatc tggccatccc
     2041 ctatgtgagc ctgaccgtga ggaggctgat tatgcacgtg gccttcttct tcctgacctt
     2101 tttctttatc gtccctatcg ccttcgtcca gtccctggct accatcgaag gaattgtcaa
     2161 agccgccccc ttcctcaagt tcattgtcga cgacaagttc atgaagagcg tgatccaggg
     2221 attcctccct ggcattgctc tgaagctgtt cctggctttt ctcccctcca tcctgatgat
     2281 catgagcaag ttcgagggct tcacctccat ctccagctta gaaaggaggg ccgccttcag
     2341 atactatatt tttaacctgg tgaatgtctt tctggcctcc gtgattgctg gagccgcctt
     2401 cgagcagctc aacagcttcc tcaaccagtc cgctaaccag attcctaaga ccatcggcgt
     2461 ggctatcccc atgaaagcca cattctttat cacatacatt atggtggacg gatgggccgg
     2521 cgtggctgga gaaatcctca tgctgaagcc tctgattatg tttcatctga aaaatgcttt
     2581 tctggtgaag acagacaagg atagggagga agccatggac cctggttcta ttggctttaa
     2641 cacaggcgag cccaggattc agctgtactt cctgctggga ctcgtctacg cccccgtgac
     2701 acccatgctg ctgcccttta tcctcgtgtt cttcgctctg gcctacattg tctacaggca
     2761 ccagatcatc aacgtgtaca atcaggagta cgaatccgcc gctgccttct ggcccgacgt
     2821 gcatggcaga gtgattgctg ccctggtgat cagccagctg ctcctcatgg gactgctcgg
     2881 aaccaagcat gccgctctgg ccgccccctt tctgattgct ctgcccgtcc tgaccatcgg
     2941 ctttcaccat ttctgcaagg gcaggtacga gcctgctttc atcaggtacc ccctgcagga
     3001 agccatgatg aaggacacct tagaaacagc cagagagcct aacctcaacc tcaagggcta
     3061 cctgcagaat gcctacgtgc accccgtgtt caagggcgac gaggatgact atgacatcga
     3121 tgacaaactg ggcaagtttg aggacgaggc tatcatcgtc cccaccaaaa ggcaatccag
     3181 gagaaacacc cctgccccca gcattatctc cggcgatgat agccctagcc tgcccttttc
     3241 cggcaagctc gtgggcaccg gtactctgga agtcctcttc cagggccctg tgagcaaggg
     3301 cgaagagctg tttacaggcg tcgtgcctat cctggtggaa ctcgatggcg acgtcaacgg
     3361 ccacaaattc tccgtgagcg gcgaaggcga aggcgacgcc acctacggaa aactcaccct
     3421 caagtttatc tgtaccacag gcaaactccc cgtcccctgg cctacactgg tgaccaccct
     3481 gacctacggc gtccaatgct tctccagata tcccgaccac atgaagcagc acgacttctt
     3541 caagtccgcc atgcctgaag gctatgtcca ggaaagaaca atctttttca aggacgacgg
     3601 aaactacaaa acaagagccg aggtgaagtt cgaaggcgac acactcgtga acagaatcga
     3661 gctgaagggc atcgacttca aagaagatgg caacatcctc ggacacaagc tcgagtataa
     3721 ttataactcc cataacgtgt atattatggc cgacaaacag aaaaatggaa tcaaggtgaa
     3781 tttcaagatt agacacaaca tcgaagacgg cagcgtgcaa ctcgccgacc attaccagca
     3841 gaacaccccc atcggagacg gacccgtgct gctgcctgat aaccactacc tgtccacaca
     3901 gtccgctctg agcaaagacc ccaacgaaaa gagggaccat atggtcctcc tggagtttgt
     3961 gaccgccgcc ggcattacac tgggcatgga cgaactgtat aaggccgctg attataagga
     4021 tgatgatgat aaatgaaagc ttaagtttaa accgctgatc agcctcgact gtgccttcta
     4081 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     4141 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     4201 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     4261 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     4321 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4381 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4441 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4501 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4561 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4621 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4681 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4741 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     4801 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4861 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4921 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4981 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5041 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5101 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     5161 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5221 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5281 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5341 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5401 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5461 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5521 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5581 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5641 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5701 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5761 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5821 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5881 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5941 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     6001 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6061 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6121 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     6181 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6241 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     6301 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6361 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6421 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6481 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6541 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6601 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6661 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6721 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6781 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6841 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6901 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6961 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7021 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7081 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7141 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7201 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt
     7261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7381 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     7441 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     7501 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7561 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7621 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7681 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7741 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7801 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7861 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7921 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7981 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8041 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8101 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8161 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8221 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8281 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     8341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     8461 cctgacgtc
//
