LOCUS       Exported                6703 bp ds-DNA     circular SYN 20-APR-2022
DEFINITION  Soluble light-dependent effector protein (HP1alpha, phosphomimetic 
            NTE mutant with deletion of C-terminal extension)for optodroplet 
            formation or recruitment to CIBN localizers.
ACCESSION   .
VERSION     .
KEYWORDS    pPHR-mCherry-nE-mHP1alpha-delCTE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6703)
  TITLE     Rippe Lab Plasmids 2022
REFERENCE   2  (bases 1 to 6703)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 20, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6703
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             625..2118
                     /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"
     CDS             2128..2835
                     /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(2273..2291)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2541..2560
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3491..3612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3528..3547)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3582..3601
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3619..4074)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3756..3777)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3968..3987
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4101..4205
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4207..4564
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4230..4250)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4415..4550
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4477..4496
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4599..5393
                     /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(4653..4672)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5263..5282
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5581..5600)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5625..5672
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6001..6589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6490..6509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc caccatgaag atggacaaaa agactatagt ttggtttaga
      661 agagacctaa ggattgagga taatcctgca ttagcagcag ctgctcacga aggatctgtt
      721 tttcctgtct tcatttggtg tcctgaagaa gaaggacagt tttatcctgg aagagcttca
      781 agatggtgga tgaaacaatc acttgctcac ttatctcaat ccttgaaggc tcttggatct
      841 gacctcactt taatcaaaac ccacaacacg atttcagcga tcttggattg tatccgcgtt
      901 accggtgcta caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt agttcgggac
      961 cataccgtaa aggagaagct ggtggaacgt gggatctctg tgcaaagcta caatggagat
     1021 ctattgtatg aaccgtggga gatatactgc gaaaagggca aaccttttac gagtttcaat
     1081 tcttactgga agaaatgctt agatatgtcg attgaatccg ttatgcttcc tcctccttgg
     1141 cggttgatgc caataactgc agcggctgaa gcgatttggg cgtgttcgat tgaagaacta
     1201 gggctggaga atgaggccga gaaaccgagc aatgcgttgt taactagagc ttggtctcca
     1261 ggatggagca atgctgataa gttactaaat gagttcatcg agaagcagtt gatagattat
     1321 gcaaagaaca gcaagaaagt tgttgggaat tctacttcac tactttctcc gtatctccat
     1381 ttcggggaaa taagcgtcag acacgttttc cagtgtgccc ggatgaaaca aattatatgg
     1441 gcaagagata agaacagtga aggagaagaa agtgcagatc tttttcttag gggaatcggt
     1501 ttaagagagt attctcggta tatatgtttc aacttcccgt ttactcacga gcaatcgttg
     1561 ttgagtcatc ttcggttttt cccttgggat gctgatgttg ataagttcaa ggcctggaga
     1621 caaggcagga ccggttatcc gttggtggat gccggaatga gagagctttg ggctaccgga
     1681 tggatgcata acagaataag agtgattgtt tcaagctttg ctgtgaagtt tcttctcctt
     1741 ccatggaaat ggggaatgaa gtatttctgg gatacacttt tggatgctga tttggaatgt
     1801 gacatccttg gctggcagta tatctctggg agtatccccg atggccacga gcttgatcgc
     1861 ttggacaatc ccgcgttaca aggcgccaaa tatgacccag aaggtgagta cataaggcaa
     1921 tggcttcccg agcttgcgag attgccaact gaatggatcc atcatccatg ggacgctcct
     1981 ttaaccgtac tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa acccattgta
     2041 gacatcgaca cagctcgtga gctactagct aaagctattt caagaacccg tgaagcacag
     2101 atcatgatcg gagcagcagc ccgggtcatg gtgagcaagg gcgaggagga taacatggcc
     2161 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     2221 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     2281 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     2341 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     2401 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     2461 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     2521 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     2581 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     2641 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     2701 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     2761 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     2821 gacgagctgt acaagatggg aaagaagacc aagaggacag ccgacgagga agaggaagag
     2881 gatgaggagg aatatgtggt ggaaaaggtg ttggacaggc gcatggttaa ggggcaagtg
     2941 gaatatctgt tgaagtggaa aggcttttct gaggagcaca atacttggga acctgagaag
     3001 aacttggatt gtcctgaact aatttctgag tttatgaaaa agtataagaa gatgaaggag
     3061 ggtgaaaaca ataagcccag ggagaaatca gaaggaaaca agaggaaatc cagtttctcc
     3121 aacagcgctg atgatattaa atctaaaaaa aagagagagc aaagcaatga tatcgctcgg
     3181 ggctttgaga gaggactgga accagaaaag atcatcggag caacagattc ctgcggtgac
     3241 ttaatgttct taatgaaatg gaaagacaca gatgaagctg acctggttct tgcaaaagaa
     3301 gctaacgtga agtgtccaca gattgtgata gcattttatg aagagagact gacgtggcac
     3361 gcatattaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     3421 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3481 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3541 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3601 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3661 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3721 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3781 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3841 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3901 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3961 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4021 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4081 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4141 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4201 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4261 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4321 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4381 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4441 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4501 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4561 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4621 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4681 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4741 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4801 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4861 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4921 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4981 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5041 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5101 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5161 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5221 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5281 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5341 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5401 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5461 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5521 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5581 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5641 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5701 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5761 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5821 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5881 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5941 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6001 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6061 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6661 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
