LOCUS       Exported                5957 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  LexATF-T2A-NLS-LexA-mCherry-LEXY construct for light control of 
            transcriptional repression.
ACCESSION   .
VERSION     .
KEYWORDS    LEXY TR (without KRAB domain) (pDN162)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5957)
  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 5957)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5957
                     /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             1437..2141
                     /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(1579..1597)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    2745..2866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2782..2801)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2836..2855
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2873..3328)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3010..3031)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3222..3241
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3355..3459
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3461..3818
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3484..3504)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3669..3804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3731..3750
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3853..4647
                     /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(3907..3926)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4517..4536
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4835..4854)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4879..4926
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5255..5843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5744..5763
                     /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 ggctctgtga
     1441 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     1501 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     1561 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     1621 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     1681 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     1741 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     1801 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     1861 agaaaaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     1921 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     1981 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     2041 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     2101 agggccgcca ctccaccggc ggcatggacg agctgtacaa gggtggatct ggaggttcag
     2161 gtggaagttt ggctactaca cttgaacgta ttgagaagaa ctttgtcatt actgacccaa
     2221 gattgccaga taatcccatt atattcgcgt ccgatagttt cttgcagttg acagaatata
     2281 gccgtgaaga aattttggga agaaactgca ggtttctaca aggtcctgaa actgatcgcg
     2341 cgacagtgag aaaaattaga gatgccatag ataaccaaac agaggtcact gttcagctga
     2401 ttaattatac aaagagtggt aaaaagttct ggaacctctt tcacttgcag cctatgcgag
     2461 atcagaaggg agatgtccag tactttattg gggttcagtt ggatggaact gagcatgtcc
     2521 gagatgctgc cgagagagag ggagtcatgc tgattaagaa aactgcagaa aatattgacg
     2581 agctgctgaa ggagctggcc gacctgaacc tggactagct cgagcggccg cgactctaga
     2641 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     2701 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     2761 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2821 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     2881 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2941 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3001 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3061 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3121 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3181 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3241 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3301 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     3361 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3421 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     3481 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3541 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3601 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3661 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3721 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3781 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     3841 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3901 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3961 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4021 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4081 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4141 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4201 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4261 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4321 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4381 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4441 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4501 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4561 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4621 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4681 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4741 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4801 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     4861 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     4921 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     4981 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5041 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5101 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5161 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5221 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5281 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5341 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5401 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5461 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5521 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5581 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5641 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5701 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5761 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5821 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5881 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5941 gtattaccgc catgcat
//
