LOCUS       Exported                5745 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 5745)
  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..5745
                     /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             643..1050
                     /codon_start=1
                     /label=PH domain
                     /translation="MSGLRSHGLQDDPDLQALLKGSQLLKVKSSSWRRERFYKLQEDCK
                     TIWQESRKVMRSPESQLFSIEDIQEVRMGHRTEGLEKFARDIPEDRCFSIVFKDQRNTL
                     DLIAPSPADAQHWVQGLRKIIHHSGSMDQRQK"
     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"
     misc_feature    2357..2409
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2533..2654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2661..3116)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3143..3247
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3249..3606
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3457..3592
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3641..4435
                     /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    4667..4714
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5043..5631
                     /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 agtgctggtg gtagtgctgg tggtagtgct
     2341 ggtggtactg gtggtcctcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc
     2401 ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt tgtagaggtt
     2461 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2521 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2581 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2641 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2701 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2761 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2821 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2881 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2941 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3001 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3061 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     3121 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3181 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3241 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3361 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3421 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3481 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3541 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3601 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3661 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3721 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3781 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     3841 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3901 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3961 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4021 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4081 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4141 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     4201 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4261 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4321 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4381 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4441 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4501 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4561 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4621 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4681 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4741 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4801 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4861 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     4921 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4981 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5041 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5101 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5161 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5221 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5281 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5341 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5401 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5461 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5521 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5581 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5641 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5701 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
