LOCUS       pcDNA3-liso-mito        7196 bp DNA     circular SYN 21-JAN-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7196)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 7, 2023 from SnapGene Viewer 7.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             913..1728
                     /codon_start=1
                     /label=TMEM192
                     /note="color: #993366"
                     /translation="TMAAGGRMEDGSLDITQSIEDDPLLDAQLLPHHSLQAHFRPRFHP
                     LPTVIIVNLLWFIHLVFVVLAFLTGVLCSYPNPNEDKCPGNYTNPLKVQTVIILGKVIL
                     WILHLLLECYIQYHHSKIRNRGYNLIYRSTRHLKRLALMIQSSGNTVLLLILCMQHSFP
                     EPGRLYLDLILAILALELICSLICLLIYTVKIRRFNKAKPEPDILEEEKIYAYPSNITS
                     ETGFRTISSLEEIVEKQGDTIEYLKRHNALLSKRLLALTSSDLGCQPSRT"
     regulatory      914..923
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             1729..1824
                     /codon_start=1
                     /label=linker long
                     /note="color: #00ffff"
                     /translation="DVGGGGSEGGGSGGPGSGGEGSAGGGSAGGGS"
     CDS             1825..1872
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /note="color: #05fd14"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1909..1965
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #cc99b2
                     Cleavage site after base 1962"
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1978..2076
                     /codon_start=1
                     /label=OMM-targeting sequence (Tom20)
                     /note="color: #3366ff"
                     /translation="MVGRNSAIAAGVCGALFIGYCIYFDRKRRSDPN"
     CDS             2077..2721
                     /codon_start=1
                     /label=GFP 1-10
                     /note="color: #339966"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGK
                     YKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEKGT"
     promoter        complement(2748..2766)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    2792..3016
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      3062..3490
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3504..3834
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3685..3820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             3901..4695
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4869..4990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5039..5055)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5063..5079
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     promoter        complement(join(5087..5093,5094..5111,5112..5117))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5087 .. 5093 / #ffffff / -10
                        2: 5094 .. 5111 / #ffffff
                        3: 5112 .. 5117 / #ffffff / -35"
     protein_bind    5132..5153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(5441..6029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(join(6200..6991,6992..7060))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 6200 .. 6991 / #ccffcc
                        2: 6992 .. 7060 / #ccffcc / signal sequence
                     Cleavage site after base 6991"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7061..7165)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccaccatggc ggcggggggc aggatggagg acggttcctt ggatatcacc
      961 cagagtattg aagacgaccc acttctggat gcccagcttc tcccacacca ctcattacaa
     1021 gctcacttta gaccccgatt ccatcctctt cctacagtca tcatagtgaa tcttctgtgg
     1081 tttattcatc tcgtgtttgt tgttttagca tttttaacag gtgtgctttg ttcttatcct
     1141 aatccaaatg aggacaagtg cccaggaaat tacacaaacc cattgaaagt tcagacggtt
     1201 ataatccttg ggaaagttat tttgtggatt ctccatttac tccttgaatg ctacatccag
     1261 tatcaccaca gcaaaatcag aaaccgaggc tataacttga tctaccgatc aacaaggcat
     1321 ctcaagagac ttgcgttgat gatacagtcc tctggcaaca cagtgcttct cctcatactg
     1381 tgcatgcagc actccttccc agagcctggc agattgtatc ttgacctcat tctggccatc
     1441 ttggcactgg aactcatctg ttccctgata tgtctcctca tttacacagt gaaaatccgg
     1501 agatttaata aagctaaacc agagcctgat atacttgaag aagaaaaaat ctatgcttac
     1561 cccagcaata ttacctcgga gactggattc agaactattt caagcctaga agaaattgtt
     1621 gaaaagcaag gagacaccat tgaatacctg aagcgacaca atgcgctgct gagtaagcga
     1681 ttgttggctc tcacttcctc agacctgggc tgtcagccaa gtagaacgga cgttggtggt
     1741 ggcggatcag aaggaggcgg tagcgggggc cctggttcgg gaggggaagg ttctgctggg
     1801 ggagggagcg ctggcggggg gtctcgggac cacatggtgc tgcacgagta cgtgaacgcc
     1861 gctggcatca caggccccgg gcttaaggag ctcgcatgcg gaagcggagc tactaacttc
     1921 agcctgctga agcaggctgg agacgtggag gagaaccctg gacctagatc tgaattcatg
     1981 gtgggccgga acagcgccat cgccgcgggc gtgtgcggtg ccctcttcat agggtactgc
     2041 atctactttg accgcaaaag gcggagtgac cccaactcca aaggagaaga actgtttacc
     2101 ggcgtggtgc caattctcgt ggaactggat ggcgatgtga atggccacaa attttctgtc
     2161 agaggagagg gtgaaggtga tgccacaatc ggaaagctca ccctgaaatt catctgcacc
     2221 actggaaagc tccctgtgcc atggccaaca ctggtcacta ccctgaccta cggcgtgcag
     2281 tgcttttcca gatacccaga ccatatgaag aggcatgact ttttcaagag cgccatgccc
     2341 gagggctatg tgcaggagag aaccatctct ttcaaagatg acgggaaata caagacccgc
     2401 gctgtggtca agttcgaagg agacacactg gtgaatagaa tcgagttgaa gggcacagac
     2461 tttaaggaag atggaaacat tctcggccac aagctggaat acaactttaa ctcccacaat
     2521 gtgtacatca cagccgacaa gcaaaagaat ggcatcaagg ctaacttcac agtcagacac
     2581 aacgtcgagg atggaagcgt gcagctggcc gaccattatc aacagaacac tccaatcggc
     2641 gacggccctg tgctcctccc agacaaccat tacctgtcca cccagacagt cctgagcaaa
     2701 gatccaaatg aaaaaggaac ataactcgag cctctagagg gccctattct atagtgtcac
     2761 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2821 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2881 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2941 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3001 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3061 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3121 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3181 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     3241 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3301 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3361 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3421 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3481 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3541 caggcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3601 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3661 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3721 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3781 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3841 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3901 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3961 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4021 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4081 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4141 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4201 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4261 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4321 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4381 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4441 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4501 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4561 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4621 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4681 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4741 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4801 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4861 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4921 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4981 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5041 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5101 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5161 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5221 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5281 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5341 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5401 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5461 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5521 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5581 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     5641 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5701 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5761 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5821 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5881 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5941 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6001 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6061 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6121 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6181 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6241 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6301 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6361 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6421 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6481 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6541 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6601 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6661 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6721 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6781 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6841 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6901 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6961 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7021 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7081 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7141 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
