LOCUS       Exported                6039 bp ds-DNA     circular SYN 20-OCT-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6039)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 20, 2022 from SnapGene 5.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6039
                     /organism="recombinant plasmid"
                     /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"
     CDS             646..660
                     /codon_start=1
                     /label=Linker
                     /translation="SGLRS"
     CDS             661..1050
                     /codon_start=1
                     /label=PLC d1
                     /translation="HGLQDDPDLQALLKGSQLLKVKSSSWRRERFYKLQEDCKTIWQES
                     RKVMRSPESQLFSIEDIQEVRMGHRTEGLEKFARDIPEDRCFSIVFKDQRNTLDLIAPS
                     PADAQHWVQGLRKIIHHSGSMDQRQK"
     regulatory      1066..1075
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1072..2016
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 703 nm (Shcherbakova et al., 2016)"
                     /label=miRFP703
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSLVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     CDS             2032..2310
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     CDS             2326..2604
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     misc_feature    2651..2703
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2827..2948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2955..3410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3437..3541
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3543..3900
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3751..3886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3935..4729
                     /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"
     polyA_signal    4961..5008
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5337..5925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 attccggcca ccatgtccgg actcagatcc
      661 cacgggctcc aggatgaccc ggaccttcag gcccttctga agggcagcca gcttctgaag
      721 gtgaagtcca gctcgtggcg tagggaacgc ttctacaagc tacaggagga ctgcaagacc
      781 atctggcagg aatctcgaaa ggtcatgagg tccccggagt cgcagctgtt ctccatcgag
      841 gacattcagg aggtacggat gggacaccgc acagaaggcc tggagaagtt tgcccgagac
      901 atccccgagg atcgatgctt ctccattgtc ttcaaggacc agcgcaacac cctagacctc
      961 attgccccat caccagctga cgctcagcac tgggtgcagg gcctgcgcaa gatcatccac
     1021 cactccggct ccatggacca gcggcagaag gcggatccac cggtcgccac catggtagca
     1081 ggtcatgcct ctggcagccc cgcattcggg accgcctctc attcgaattg cgaacatgaa
     1141 gagatccacc tcgccggctc gatccagccg catggcgcgc ttctggtcgt cagcgaacat
     1201 gatcatcgcg tcatccaggc cagcgccaac gccgcggaat ttctgaatct cggaagcgta
     1261 ctcggcgttc cgctcgccga gatcgacggc gatctgttga tcaagatcct gccgcatctc
     1321 gatcccaccg ccgaaggcat gccggtcgcg gtgcgctgcc ggatcggcaa tccctctacg
     1381 gagtactgcg gtctgatgca tcggcctccg gaaggcgggc tgatcatcga actcgaacgt
     1441 gccggcccgt cgatcgatct gtcaggcacg ctggcgccgg cgctggagcg gatccgcacg
     1501 gcgggttcac tgcgcgcgct gtgcgatgac accgtgctgc tgtttcagca gtgcaccggc
     1561 tacgaccggg tgatggtgta tcgtttcgat gagcaaggcc acggcctggt attctccgag
     1621 tgccatgtgc ctgggctcga atcctatttc ggcaaccgct atccgtcgtc gctggtcccg
     1681 cagatggcgc ggcagctgta cgtgcggcag cgcgtccgcg tgctggtcga cgtcacctat
     1741 cagccggtgc cgctggagcc gcggctgtcg ccgctgaccg ggcgcgatct cgacatgtcg
     1801 ggctgcttcc tgcgctcgat gtcgccgatc catctgcagt tcctgaagga catgggcgtg
     1861 cgcgccaccc tggcggtgtc gctggtggtc ggcggcaagc tgtggggcct ggttgtctgt
     1921 caccattatc tgccgcgctt catccgtttc gagctgcggg cgatctgcaa acggctcgcc
     1981 gaaaggatcg cgacgcggat caccgcgctt gagagcaagg gagcaagtgg aatcctctgg
     2041 catgagatgt ggcatgaagg cctggaagag gcatctcgtt tgtactttgg ggaaaggaac
     2101 gtgaaaggca tgtttgaggt gctggagccc ttgcatgcta tgatggaacg gggcccccag
     2161 actctgaagg aaacatcctt taatcaggcc tatggtcgag atttaatgga ggcccaagag
     2221 tggtgcagga agtacatgaa atcagggaat gtcaaggacc tcctccaagc ctgggacctc
     2281 tattatcatg tgttccgacg aatctcaaag aagggagcaa gtggaatcct ctggcatgag
     2341 atgtggcatg aaggcctgga agaggcatct cgtttgtact ttggggaaag gaacgtgaaa
     2401 ggcatgtttg aggtgctgga gcccttgcat gctatgatgg aacggggccc ccagactctg
     2461 aaggaaacat cctttaatca ggcctatggt cgagatttaa tggaggccca agagtggtgc
     2521 aggaagtaca tgaaatcagg gaatgtaaag gacctcctcc aagcctggga cctctattat
     2581 catgtgttcc gacgaatctc aaagagtgct ggtggtagtg ctggtggtag tgctggtggt
     2641 actggtggtc ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga
     2701 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     2761 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2821 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2881 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2941 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3001 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3061 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3121 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3181 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3241 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3301 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3361 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3421 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3481 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3541 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3601 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3661 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3721 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3781 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3841 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3901 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3961 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4021 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4081 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4141 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4201 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4261 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4321 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4381 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4441 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4501 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4561 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4621 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4681 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4741 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4801 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4861 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4921 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4981 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5041 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5101 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5161 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5221 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5281 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5341 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5401 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5461 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5521 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5581 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5641 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5701 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5761 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5821 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5881 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5941 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6001 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
