LOCUS       Exported                6122 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expression of light-regulated p38 inhibitor, firefly 
            luciferase-fused, dark-state mutant photosensor (AsLov2Ja.C450A), 
            inhibitor MK3BD3-13F.
ACCESSION   .
VERSION     .
KEYWORDS    pLuc-Optop38i-dsm
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6122)
  AUTHORS   Melero-Fernandez de Mera RM, Li LL, Popinigis A, Cisek K, Tuittila 
            M, Yadav L, Serva A, Courtney MJ
  TITLE     A simple optogenetic MAPK inhibitor design reveals resonance between
            transcription-regulating circuitry and temporally-encoded inputs.
  JOURNAL   Nat Commun. 2017 May 12;8:15017. doi: 10.1038/ncomms15017.
  PUBMED    28497795
REFERENCE   2  (bases 1 to 6122)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6122
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        130..433
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        434..637
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     588..608
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      676..685
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             682..2331
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(748..766)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2979..3100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3016..3035)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3070..3089
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3107..3562)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3244..3265)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3456..3475
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3589..3693
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3695..4052
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3718..3738)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3903..4038
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3965..3984
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4087..4881
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4141..4160)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4751..4770
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5069..5088)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5113..5160
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5489..6077
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5978..5997
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
       61 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      121 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      181 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      241 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      301 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      361 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      421 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      481 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      541 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      601 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
      661 ctagcgctac cggtcgccac catggaagac gccaaaaaca taaagaaagg cccggcgcca
      721 ttctatccgc tggaagatgg aaccgctgga gagcaactgc ataaggctat gaagagatac
      781 gccctggttc ctggaacaat tgcttttaca gatgcacata tcgaggtgga catcacttac
      841 gctgagtact tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata tgggctgaat
      901 acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc aattctttat gccggtgttg
      961 ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa
     1021 ttgctcaaca gtatgggcat ttcgcagcct accgtggtgt tcgtttccaa aaaggggttg
     1081 caaaaaattt tgaacgtgca aaaaaagctc ccaatcatcc aaaaaattat tatcatggat
     1141 tctaaaacgg attaccaggg atttcagtcg atgtacacgt tcgtcacatc tcatctacct
     1201 cccggtttta atgaatacga ttttgtgcca gagtccttcg atagggacaa gacaattgca
     1261 ctgatcatga actcctctgg atctactggt ctgcctaaag gtgtcgctct gcctcataga
     1321 actgcctgcg tgagattctc gcatgccaga gatcctattt ttggcaatca aatcattccg
     1381 gatactgcga ttttaagtgt tgttccattc catcacggtt ttggaatgtt tactacactc
     1441 ggatatttga tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga agagctgttt
     1501 ctgaggagcc ttcaggatta caagattcaa agtgcgctgc tggtgccaac cctattctcc
     1561 ttcttcgcca aaagcactct gattgacaaa tacgatttat ctaatttaca cgaaattgct
     1621 tctggtggcg ctcccctctc taaggaagtc ggggaagcgg ttgccaagag gttccatctg
     1681 ccaggtatca ggcaaggata tgggctcact gagactacat cagctattct gattacaccc
     1741 gagggggatg ataaaccggg cgcggtcggt aaagttgttc cattttttga agcgaaggtt
     1801 gtggatctgg ataccgggaa aacgctgggc gttaatcaaa gaggcgaact gtgtgtgaga
     1861 ggtcctatga ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc cttgattgac
     1921 aaggatggat ggctacattc tggagacata gcttactggg acgaagacga acacttcttc
     1981 atcgttgacc gcctgaagtc tctgattaag tacaaaggct atcaggtggc tcccgctgaa
     2041 ttggaatcca tcttgctcca acaccccaac atcttcgacg caggtgtcgc aggtcttccc
     2101 gacgatgacg ccggtgaact tcccgccgcc gttgttgttt tggagcacgg aaagacgatg
     2161 acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa aaagttgcgc
     2221 ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga
     2281 aaaatcagag agatcctcat aaaggccaag aagggcggaa agatcgccgt gcggtacaag
     2341 tccggactca gatctcgagc tcaagcttcg aattcactgg caacaacact ggagagaatc
     2401 gagaaaaact tcgtcatcac cgaccctaga ctgcccgata atcctatcat cttcgcctcc
     2461 gactcatttc tccagctcac tgagtactct cgggaggaaa ttctgggccg gaatgcccgg
     2521 ttcctccagg gacctgaaac cgatagagcc actgtccgaa aaatccggga tgccatcgac
     2581 aatcagactg aggtgaccgt ccagctcatc aattacacca aatctggcaa aaaattctgg
     2641 aacctgttcc acctccagcc tatgagagat cagaagggcg acgtccagta ctttattggc
     2701 gtccagctcg atggaactga acatgtgagg gacgctgcag aacgagaggg cgtgatgctc
     2761 atcaaaaaaa ccgccgaaaa catcgatgag gccgctaaag agctgtctag aaagcggaag
     2821 aaggacctgc ggatctcctg catgttctag gatccaccgg atctagataa ctgatcataa
     2881 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2941 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3001 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3061 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     3121 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3181 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3241 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3301 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3361 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3421 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3481 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3541 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3601 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3661 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3721 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3781 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3841 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3901 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3961 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4021 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4081 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4141 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4201 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4261 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4321 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4381 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4441 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4501 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4561 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4621 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4681 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4741 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4801 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4861 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4921 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4981 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5041 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5101 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5161 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5221 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5281 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5341 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5401 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5461 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5521 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5581 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5641 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5701 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5761 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5821 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5881 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5941 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6001 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6061 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6121 gg
//
