LOCUS       Exported                7156 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian Expression, Bacterial Expression.
ACCESSION   .
VERSION     .
KEYWORDS    mCherry-CRY2-5ptaseOCRL
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7156)
  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 7156)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7156
                     /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    3944..4065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3981..4000)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4035..4054
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4072..4527)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4209..4230)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4421..4440
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4554..4658
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4660..5017
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4683..4703)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4868..5003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4930..4949
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5052..5846
                     /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(5106..5125)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5716..5735
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6034..6053)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6078..6125
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6454..7042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6943..6962
                     /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 gtcgatcgta
     2881 tgtcaacatt cagactttca gattttttgt tggaacttgg aatgtgaatg gccagtctcc
     2941 agatagcggg ttagaacctt ggctgaactg tgatcccaat cctcctgata tctactgcat
     3001 tggattccaa gaactggact tgagcacaga agccttcttc tactttgaat ctgtgaagga
     3061 acaagaatgg tccatggctg tagagagagg tttgcattcc aaagccaagt ataagaaagt
     3121 tcaactggtg cgccttgttg ggatgatgct tcttatattt gccagaaagg atcagtgtcg
     3181 atacattcgt gatattgcta cagaaacagt tggaactgga atcatgggga aaatgggaaa
     3241 caaaggtggg gtagctgtga gatttgtatt tcacaacacc accttttgca ttgtcaattc
     3301 ccatctggct gcacacgtgg aggactttga gagaaggaat caagattata aggacatttg
     3361 tgcgagaatg agttttgtgg tcccaaatca gaccctcccg cagttgaaca tcatgaaaca
     3421 tgaggttgtc atttggttgg gagatttgaa ttatagactt tgcatgcctg atgccaatga
     3481 ggtgaaaagt cttattaata agaaagacct tcagagactc ttgaaattcg accagctaaa
     3541 tattcagcgc acacagaaaa aagcttttgt tgacttcaat gaaggggaaa tcaagttcat
     3601 ccccacttat aagtatgact ctaaaacaga ccggtgggat tccagtggga aatgccgggt
     3661 tccagcctgg tgtgaccgaa ttctttggag aggaacaaat gttaatcagc ttaattatcg
     3721 gagtcacatg gaactgaaaa ccagcgacca caagcctgtt agcgccctct tccatattgg
     3781 ggtgaaggtt gtggattgag gtaccgcggg cccgggatcc accggatcta gataactgat
     3841 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3901 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3961 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4021 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     4081 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4141 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     4201 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4261 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4321 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4381 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4441 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4501 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4561 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4621 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     4681 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4741 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4801 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4861 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4921 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4981 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     5041 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5101 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5161 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5221 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5281 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5341 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5401 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5461 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5521 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     5581 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5641 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5701 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5761 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5821 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5881 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5941 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6001 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     6061 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     6121 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     6181 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     6241 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     6301 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6361 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6421 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6481 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6541 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6601 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6661 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6721 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6781 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6841 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6901 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6961 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7021 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7081 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7141 tattaccgcc atgcat
//
