LOCUS       Exported                5799 bp ds-DNA     circular SYN 20-OCT-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5799)
  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..5799
                     /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"
     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..1776
                     /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
                     EGRHSTGGMDELY"
     CDS             1792..2070
                     /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             2086..2364
                     /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    2411..2463
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2587..2708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2715..3170)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3197..3301
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3303..3660
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3511..3646
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3695..4489
                     /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    4721..4768
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5097..5685
                     /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 catggtgagc
     1081 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     1141 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1201 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     1261 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     1321 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     1381 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     1441 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     1501 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     1561 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     1621 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     1681 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     1741 ggccgccact ccaccggcgg catggacgag ctgtacaagg gagcaagtgg aatcctctgg
     1801 catgagatgt ggcatgaagg cctggaagag gcatctcgtt tgtactttgg ggaaaggaac
     1861 gtgaaaggca tgtttgaggt gctggagccc ttgcatgcta tgatggaacg gggcccccag
     1921 actctgaagg aaacatcctt taatcaggcc tatggtcgag atttaatgga ggcccaagag
     1981 tggtgcagga agtacatgaa atcagggaat gtcaaggacc tcctccaagc ctgggacctc
     2041 tattatcatg tgttccgacg aatctcaaag aagggagcaa gtggaatcct ctggcatgag
     2101 atgtggcatg aaggcctgga agaggcatct cgtttgtact ttggggaaag gaacgtgaaa
     2161 ggcatgtttg aggtgctgga gcccttgcat gctatgatgg aacggggccc ccagactctg
     2221 aaggaaacat cctttaatca ggcctatggt cgagatttaa tggaggccca agagtggtgc
     2281 aggaagtaca tgaaatcagg gaatgtaaag gacctcctcc aagcctggga cctctattat
     2341 catgtgttcc gacgaatctc aaagagtgct ggtggtagtg ctggtggtag tgctggtggt
     2401 actggtggtc ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga
     2461 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     2521 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2581 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2641 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2701 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     2761 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2821 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2881 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2941 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3001 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3061 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3121 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3181 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3241 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3301 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3361 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3421 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3481 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3541 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3601 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3661 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     3721 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     3781 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     3841 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     3901 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     3961 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4021 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4081 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4141 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4201 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4261 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4321 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4381 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4441 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     4501 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4561 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4621 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     4681 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4741 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     4801 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     4861 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     4921 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     4981 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5041 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5101 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5161 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5221 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5281 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5341 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5401 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5461 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5521 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5581 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5641 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5701 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5761 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
