LOCUS       Exported                6826 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian Expression, Bacterial Expression.
ACCESSION   .
VERSION     .
KEYWORDS    mCherry-CRY2-iSH2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6826)
  AUTHORS   Idevall-Hagren O, Dickson EJ, Hille B, Toomre DK, De Camilli P
  TITLE     Optogenetic control of phosphoinositide metabolism.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Aug 28;109(35):E2316-23. doi: 
            10.1073/pnas.1211305109. Epub 2012 Jul 30.
  PUBMED    22847441
REFERENCE   2  (bases 1 to 6826)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6826
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        48..351
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        352..555
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     506..526
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      584..593
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             590..1297
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(735..753)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1003..1022
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1343..2836
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     polyA_signal    3614..3735
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3651..3670)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3705..3724
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3742..4197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3879..3900)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4091..4110
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4224..4328
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4330..4687
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4353..4373)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4538..4673
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4600..4619
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4722..5516
                     /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(4776..4795)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5386..5405
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5704..5723)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5748..5795
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6124..6712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6613..6632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
       61 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      121 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      181 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      241 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      301 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      361 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      421 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      481 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      541 tatataagca gagctggttt agtgaaccgt cagatccgct agcgccacca tggtgagcaa
      601 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
      661 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
      721 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
      781 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
      841 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
      901 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
      961 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     1021 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     1081 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     1141 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     1201 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     1261 ccgccactcc accggcggca tggacgagct gtacaagaga tctcgaagcg cggccgcggg
     1321 agcaggagga gcagctcgag ctatgaagat ggacaaaaag actatagttt ggtttagaag
     1381 agacctaagg attgaggata atcctgcatt agcagcagct gctcacgaag gatctgtttt
     1441 tcctgtcttc atttggtgtc ctgaagaaga aggacagttt tatcctggaa gagcttcaag
     1501 atggtggatg aaacaatcac ttgctcactt atctcaatcc ttgaaggctc ttggatctga
     1561 cctcacttta atcaaaaccc acaacacgat ttcagcgatc ttggattgta tccgcgttac
     1621 cggtgctaca aaagtcgtct ttaaccacct ctatgatcct gtttcgttag ttcgggacca
     1681 taccgtaaag gagaagctgg tggaacgtgg gatctctgtg caaagctaca atggagatct
     1741 attgtatgaa ccgtgggaga tatactgcga aaagggcaaa ccttttacga gtttcaattc
     1801 ttactggaag aaatgcttag atatgtcgat tgaatccgtt atgcttcctc ctccttggcg
     1861 gttgatgcca ataactgcag cggctgaagc gatttgggcg tgttcgattg aagaactagg
     1921 gctggagaat gaggccgaga aaccgagcaa tgcgttgtta actagagctt ggtctccagg
     1981 atggagcaat gctgataagt tactaaatga gttcatcgag aagcagttga tagattatgc
     2041 aaagaacagc aagaaagttg ttgggaattc tacttcacta ctttctccgt atctccattt
     2101 cggggaaata agcgtcagac acgttttcca gtgtgcccgg atgaaacaaa ttatatgggc
     2161 aagagataag aacagtgaag gagaagaaag tgcagatctt tttcttaggg gaatcggttt
     2221 aagagagtat tctcggtata tatgtttcaa cttcccgttt actcacgagc aatcgttgtt
     2281 gagtcatctt cggtttttcc cttgggatgc tgatgttgat aagttcaagg cctggagaca
     2341 aggcaggacc ggttatccgt tggtggatgc cggaatgaga gagctttggg ctaccggatg
     2401 gatgcataac agaataagag tgattgtttc aagctttgct gtgaagtttc ttctccttcc
     2461 atggaaatgg ggaatgaagt atttctggga tacacttttg gatgctgatt tggaatgtga
     2521 catccttggc tggcagtata tctctgggag tatccccgat ggccacgagc ttgatcgctt
     2581 ggacaatccc gcgttacaag gcgccaaata tgacccagaa ggtgagtaca taaggcaatg
     2641 gcttcccgag cttgcgagat tgccaactga atggatccat catccatggg acgctccttt
     2701 aaccgtactc aaagcttctg gtgtggaact cggaacaaac tatgcgaaac ccattgtaga
     2761 catcgacaca gctcgtgagc tactagctaa agctatttca agaacccgtg aagcacagat
     2821 catgatcgga gcagcagctc gaggcgcagc agcgggagca ggaggagcag gtcgatcgct
     2881 ggatgtgaag ttgctctacc cagtgtctaa ataccagcag gatcaagttg tcaaagaaga
     2941 taatattgaa gctgtaggga aaaaattaca tgaatataat actcaatttc aagaaaaaag
     3001 tcgggaatat gatagattat atgaggaata cactcgtact tcccaggaaa tacagatgaa
     3061 aagaacagct atcgaagcat ttaatgaaac cataaaaata ttcgaagagc agtgccaaac
     3121 ccaggagcgg tacagcaaag aatacataga gaagtttaaa cgcgaaggca acgagaagga
     3181 aattcaaagg ataatgcata atcacgataa gctgaagtct cggatcagcg agatcatcga
     3241 cagcaggagg agactggagg aagacttgaa gaagcaggca gccgagtatc gcgagatcga
     3301 caagcgcatg aatagcatta agccggacct cattcagctg agaaagacaa gagatcaata
     3361 cttgatgtgg ctgacgcaga aaggggtccg gcagaagaag ctgaacgaat ggttgggaaa
     3421 cgaaaacaca gaagaccaat actcactggt agaagatgat gaggattgag gtaccgcggg
     3481 cccgggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     3541 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     3601 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3661 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3721 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3781 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3841 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3901 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3961 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     4021 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     4081 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     4141 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     4201 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4261 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4321 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     4381 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4441 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4501 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4561 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4621 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4681 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4741 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4801 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4861 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     4921 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4981 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5041 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5101 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5161 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5221 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     5281 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5341 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5401 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5461 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5521 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5581 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5641 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     5701 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     5761 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     5821 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5881 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5941 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     6001 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6061 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6121 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6181 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6241 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     6301 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6361 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6421 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6481 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6541 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6601 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6661 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6721 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6781 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
