LOCUS       Exported                6206 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  LexATF-T2A-NLS-LexA-KRAB-mCherry-LEXY construct for light control of
            transcriptional repression.
ACCESSION   .
VERSION     .
KEYWORDS    LEXY TR (with KRAB domain) (pDN163)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6206)
  AUTHORS   Niopek D, Wehler P, Roensch J, Eils R, Di Ventura B
  TITLE     Optogenetic control of nuclear protein export.
  JOURNAL   Nat Commun. 2016 Feb 8;7:10624. doi: 10.1038/ncomms10624.
  PUBMED    26853913
REFERENCE   2  (bases 1 to 6206)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6206
                     /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             1065..1118
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1467..1661
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             1686..2390
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1828..1846)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    2994..3115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3031..3050)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3085..3104
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3122..3577)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3259..3280)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3471..3490
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3604..3708
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3710..4067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3733..3753)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3918..4053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3980..3999
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4102..4896
                     /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(4156..4175)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4766..4785
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5084..5103)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5128..5175
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5504..6092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5993..6012
                     /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 atctcgagct caagcttcga attccgccac catgaaagct ctgacagcca
      661 ggcagcagga agtgttcgat ctgattagag accacatatc acagaccggt atgccaccta
      721 caagggcaga gattgcacag cgccttggtt ttcgctcacc gaatgccgcc gaagaacacc
      781 tcaaggctct tgcacgcaag ggcgttatcg agattgtgtc tggggcctcc agaggcatca
      841 ggctcctcca ggaagaggaa gagggcctcc ccttggtcgg gcgcgtggca gctggtgagc
      901 cagggggctc cggaggaagt gagagcggag gccccgctga cgccctggac gactttgacc
      961 tggatatgct tcccgcagat gccctggacg actttgactt ggacatgctg cccgcagatg
     1021 cgctggatga ctttgatctg gacatgctgc ctggtggtag cggcgaaggt cgagggagcc
     1081 tgctaacatg tggcgatgtt gaggagaacc ctggtcccgg ggccgccgcc aagagggtta
     1141 agctcgacgg cggtagtatg aaggccctta ctgcaaggca gcaagaggtc tttgacctga
     1201 ttcgagacca tatttcccag acaggtatgc ccccgacgag ggcagagatt gcacagcggc
     1261 tgggattccg gtccccgaat gctgcggagg aacacttgaa ggccctggcc cgaaaaggtg
     1321 tgattgagat tgtctcagga gcatctcggg gcataagatt gctgcaggag gaagaggagg
     1381 gccttcctct tgtaggacgc gtagccgctg gggaacccgg aggatccggc ggctctatgg
     1441 acgcaaagtc tctgacagcc tggagccgga ccttggtgac cttcaaagat gtgttcgtgg
     1501 acttcacccg ggaggagtgg aaactcttgg acaccgccca gcagatcgtt tatagaaacg
     1561 tgatgctgga gaactacaaa aaccttgtga gcttgggcta ccaactcaca aaacccgacg
     1621 taatcctgcg gctggagaag ggggaagaac catggcttgt tggcggcagc ggcggcagcg
     1681 gcggcgtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     1741 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     1801 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     1861 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     1921 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     1981 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     2041 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     2101 taatgcagaa gaaaaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     2161 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2221 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     2281 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2341 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag ggtggatctg
     2401 gaggttcagg tggaagtttg gctactacac ttgaacgtat tgagaagaac tttgtcatta
     2461 ctgacccaag attgccagat aatcccatta tattcgcgtc cgatagtttc ttgcagttga
     2521 cagaatatag ccgtgaagaa attttgggaa gaaactgcag gtttctacaa ggtcctgaaa
     2581 ctgatcgcgc gacagtgaga aaaattagag atgccataga taaccaaaca gaggtcactg
     2641 ttcagctgat taattataca aagagtggta aaaagttctg gaacctcttt cacttgcagc
     2701 ctatgcgaga tcagaaggga gatgtccagt actttattgg ggttcagttg gatggaactg
     2761 agcatgtccg agatgctgcc gagagagagg gagtcatgct gattaagaaa actgcagaaa
     2821 atattgacga gctgctgaag gagctggccg acctgaacct ggactagctc gagcggccgc
     2881 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2941 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3001 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3061 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     3121 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3181 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3241 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3301 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3361 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3421 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3481 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3541 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3601 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3661 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3721 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3781 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3841 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3901 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3961 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4021 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     4081 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4141 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4201 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4261 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4321 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4381 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4441 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4501 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4561 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4621 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4681 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4741 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4801 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4861 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4921 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4981 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5041 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     5101 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5161 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     5221 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5281 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5341 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5401 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5461 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5521 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5581 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5641 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5701 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5761 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5821 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5881 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5941 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6001 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6061 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6121 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6181 tggataaccg tattaccgcc atgcat
//
