LOCUS       Exported                7822 bp ds-DNA     circular SYN 25-JUL-2022
DEFINITION  Optically controlled eDrcKIT expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    eDrcKIT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7822)
  TITLE     HOLD - Verkhusha lab plasmids 2022
REFERENCE   2  (bases 1 to 7822)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 25, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7822
                     /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"
     sig_peptide     597..659
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     CDS             660..1361
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="SKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFED
                     GGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGE
                     IKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEG
                     RHSTGGMDELYK"
     primer_bind     complement(799..817)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1067..1086
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4610..4731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4647..4666)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4701..4720
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4738..5193)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4875..4896)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5087..5106
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5220..5324
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5326..5683
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(5349..5369)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5534..5669
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5596..5615
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5718..6512
                     /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(5772..5791)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6382..6401
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6700..6719)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6744..6791
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7120..7708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7609..7628
                     /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 gctagcatgg
      601 agacagacac actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca
      661 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
      721 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
      781 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
      841 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
      901 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
      961 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     1021 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     1081 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     1141 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     1201 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     1261 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     1321 agggccgcca ctccaccggc ggcatggacg agctgtacaa ggaattcagt gctggtagtg
     1381 ctggtagtgc tggcaccggt atgagccggg acccgttgcc cttttttcca ccgctttacc
     1441 ttggtggccc ggaaattacc accgagaact gcgagcgcga gccgattcat attcccggca
     1501 gcatccagcc gcacggcgcc ctgctcactg ccgacgggca cagcggcgag gtgctccaga
     1561 tgagcctcaa cgcggccact tttctgggac aggaacccac agtgctgcgc ggacagaccc
     1621 tcgccgcact gctgcccgag cagtggcccg cgctgcaagc ggccctgccc cccggctgcc
     1681 ccgacgccct gcaataccgc gcaacgctgg actggcctgc cgccgggcac ctttcgctga
     1741 cggtgcaccg ggtcggcgag ttgctgattc tggagttcga gccgacggag gcctgggaca
     1801 gcaccgggcc gcacgcgctg cgcaacgcga tgttcgcgct cgaaagtgcc cccaacctgc
     1861 gggcgctggc cgaggtggcg acccagacgg tccgcgagct gacgggcttt gaccgggtga
     1921 tgctctacaa atttgccccc gacgccaccg gcgaagtgat tgccgaggcc cgccgtgagg
     1981 ggctgcacgc ctttctgggc caccgttttc ccgcgtcgga cattccggcg caggcccgcg
     2041 cgctctacac ccggcacctg ctgcgcctga ccgccgacac ccgcgccgcc gccgtgccgc
     2101 tcgatcccgt cctcaacccg cagacgaatg cgcccacccc gctgggcggc gccgtgctgc
     2161 gcgccacctc gcccatgcac atgcagtacc tgcggaacat gggcgtcggg tcgagcctgt
     2221 cggtgtcggt ggtggtcggc ggccagctct ggggcctgat cgcctgccac caccagacgc
     2281 cctacgtgtt gccgcccgac ctgcgaacca cgctcgaata cctgggccgc ttgctgagcc
     2341 tgcaagttca ggtcaaggaa gcggcggacg tggcggcctt tcgccagagc ctgcgggagc
     2401 accacgcgcg ggtggccctc gcggcggcgc actcgctctc gccgcacgac accctcagtg
     2461 acccggcgct tgacctgctg ggcctgatgc gggccggggg cctgattctg cgtttcgagg
     2521 gccgctggca gacgttgggt gaagtgccgc ctgccccggc ggtggacgcg ctgctggcgt
     2581 ggctcgaaac ccagccgggc gccctggtcc agaccgacgc gctaggccaa ctgtggcccg
     2641 ccggcgccga tctcgccccc agcgcagcgg gcctgctcgc catcagcgtg ggcgagggct
     2701 ggtcggagtg cctcgtctgg ctgcggcccg aactgcggct ggaggtcgcc tggggcgggg
     2761 ccactcctga ccaggcgaaa gacgacctcg ggccgcgcca ctcattcgac acctacctcg
     2821 aagaaaaacg cggctacgcc gagccctggc atcccggcga aatcgaggag gcgcaggatc
     2881 tacgtgacac attgaccggg gcgctgggcg agcagctcga gtccatcgtc tctgcggtgg
     2941 ttggcattct gctggtcgtg gtcttggggg tggtctttgg gatcctctct agaaaatatt
     3001 tacagaaacc catgtatgaa gtacagtgga aggttgttga ggagataaat ggaaacaatt
     3061 atgtttacat agacccaaca caacttcctt atgatcacaa atgggagttt cccagaaaca
     3121 ggctgagttt tgggaaaacc ctgggtgctg gagctttcgg gaaggttgtt gaggcaactg
     3181 cttatggctt aattaagtca gatgcggcca tgactgtcgc tgtaaagatg ctcaagccga
     3241 gtgcccattt gacagaacgg gaagccctca tgtctgaact caaagtcctg agttaccttg
     3301 gtaatcacat gaatattgtg aatctacttg gagcctgcac cattggaggg cccaccctgg
     3361 tcattacaga atattgttgc tatggtgatc ttttgaattt tttgagaaga aaacgtgatt
     3421 catttatttg ttcaaagcag gaagatcatg cagaagctgc actttataag aatcttctgc
     3481 attcaaagga gtcttcctgc agcgatagta ctaatgagta catggacatg aaacctggag
     3541 tttcttatgt tgtcccaacc aaggccgaca aaaggagatc tgtgagaata ggctcataca
     3601 tagaaagaga tgtgactccc gccatcatgg aggatgacga gttggcccta gacttagaag
     3661 acttgctgag cttttcttac caggtggcaa agggcatggc tttcctcgcc tccaagaatt
     3721 gtattcacag agacttggca gccagaaata tcctccttac tcatggtcgg atcacaaaga
     3781 tttgtgattt tggtctagcc agagacatca agaatgattc taattatgtg gttaaaggaa
     3841 acgctcgact acctgtgaag tggatggcac ctgaaagcat tttcaactgt gtatacacgt
     3901 ttgaaagtga cgtctggtcc tatgggattt ttctttggga gctgttctct ttaggaagca
     3961 gcccctatcc tggaatgccg gtcgattcta agttctacaa gatgatcaag gaaggcttcc
     4021 ggatgctcag ccctgaacac gcacctgctg aaatgtatga cataatgaag acttgctggg
     4081 atgcagatcc cctaaaaaga ccaacattca agcaaattgt tcagctaatt gagaagcaga
     4141 tttcagagag caccaatcat atttactcca acttagcaaa ctgcagcccc aaccgacaga
     4201 agcccgtggt agaccattct gtgcggatca attctgtcgg cagcaccgct tcctcctccc
     4261 agcctctgct tgtgcacgac gatgtctcta gaggcggtgg tggcagcgga ggagggggga
     4321 gcggcggcgg tggcagtggc ggcggcggta gtcgcagccg ctttgtgaaa aaagatagcg
     4381 ccggcagtgc gggtagtgcg gggagcgccg gtagctatct ggcgaacgaa attctgtggg
     4441 gcagcgctgg tagcgcagga agcgccggca gtgcagggtt ttgctatgaa aacgaagtgg
     4501 cgctgtcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     4561 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     4621 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4681 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     4741 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     4801 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     4861 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     4921 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     4981 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5041 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5101 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     5161 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     5221 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5281 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     5341 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     5401 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     5461 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     5521 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5581 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5641 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     5701 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     5761 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     5821 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     5881 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5941 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     6001 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     6061 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     6121 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     6181 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     6241 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     6301 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     6361 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     6421 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     6481 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     6541 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     6601 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6661 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     6721 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     6781 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     6841 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     6901 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     6961 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7021 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7081 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7141 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7201 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7261 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7321 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7381 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7441 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7501 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7561 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7621 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     7681 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     7741 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     7801 taaccgtatt accgccatgc at
//
