LOCUS       sequence-368792-        6580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  original vector to make all constructs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  AUTHORS   Cho SI, Lee S, Mok YG, Lim K, Lee J, Lee JM, Chung E, Kim JS
  TITLE     Targeted A-to-G base editing in human mitochondrial DNA with 
            programmable deaminases.
  JOURNAL   Cell. 2022 May 12;185(10):1764-1776.e12. doi: 
            10.1016/j.cell.2022.03.039. Epub 2022 Apr 25.
  PUBMED    35472302
REFERENCE   2  (bases 1 to 6580)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2022.03.039"; journalName: "Cell"; date: 
            "2022-05-12- 12"; volume: "185"; issue: "10"; pages: "1764-1776.e12"
COMMENT     Sequence Label: TALED_Right-stuffer-1397C-8e
FEATURES             Location/Qualifiers
     source          1..6580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             885..962
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MASVLTPLLLRGLTGSARRLPVPRAK"
     CDS             975..1040
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1466..2140
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1466..2140
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1596..1614)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1864..1883
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2058..2081
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             join(2182..2184,2185..2898)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2182..2184,2185..2895)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2224..2245)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2485..2504)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2832..2853
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3792..3809)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3798..4022
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4253..4374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4290..4309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4344..4363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4423..4439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4423..4439)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4436..4458)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4447..4463
                     /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)."
     promoter        complement(join(4471..4477,4478..4495,4496..4501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4516..4537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4654..4671)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4825..5413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4905..4924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5584..6375,6376..6444))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6207..6226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6445..6549)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 ggagatggcc tctgtcttga
      901 caccactcct gctcaggggc ctgactggat cagctaggcg gctccccgtt cctcgcgcaa
      961 aaatacacag cctggactat aaggaccatg acggtgatta caaagatcac gacatagatt
     1021 ataaggatga tgatgataag gggatccgaa ttcaagatct acgcacgctc ggctacagcc
     1081 agcagcaaca ggagaagatc aaaccgaagg ttcgttcgac agtggcgcag caccacgagg
     1141 cactggtcgg ccatgggttt acacacgcgc acatcgttgc gctcagccaa cacccggcag
     1201 cgttagggac cgtcgctgtc aagtatcagg acatgatcgc agcgttgcca gaggcgacac
     1261 acgaagcgat cgttggcgtc ggcaaacagt ggtccggcgc acgcgctctg gaggccttgc
     1321 tcacggtggc gggagagttg agaggtccac cgttacagtt ggacacaggc caacttctca
     1381 agattgcaaa acgtggcggc gtgaccgcag tggaggcagt gcatgcatgg cgcaatgcac
     1441 tgacgggtgc ccccctgagg agacgatggc ctcctccgag gacgtcatca aggagttcat
     1501 gcgcttcaag gtgcgcatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     1561 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg
     1621 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc cagtacggct ccaaggccta
     1681 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     1741 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     1801 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     1861 cggccccgta atgcagaaga agaccatggg ctgggaggcc tccaccgagc ggatgtaccc
     1921 cgaggacggc gccctgaagg gcgagatcaa gatgaggctg aagctgaagg acggcggcca
     1981 ctacgacgcc gaggtcaaga ccacctacat ggccaagaag cccgtgcagc tgcccggcgc
     2041 ctacaagacc gacatcaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     2101 acagtacgag cgcgccgagg gccgccactc caccggcgcc gaattcatga catcaattat
     2161 tatacatcgg aggaggatcc agtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2221 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2281 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2341 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     2401 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2461 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2521 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2581 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2641 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     2701 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2761 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     2821 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2881 gacgagctgt acaagtaacg tctcccggtc tgactccgga acaggtggtg gcgattgcaa
     2941 gcaacggcgg cggcaaacag gctctagaga gcattgttgc ccagctctcc agacctgatc
     3001 cggcgctagc cgcgttgaca aatgatcatc ttgttgctct tgcttgtctt ggcggacgac
     3061 ctgccctgga tgctgtgaaa aaggggttgc tagtcggcag tgccatacct gtgaagcggg
     3121 gagcaacagg ggagacaaag gtgttcacag gcaactctaa cagtccaaag agccccacca
     3181 aaggcgggtg ttcaggaagc gaaactcctg gtacctcaga gtccgctact cccgaatcct
     3241 ctgaggtgga gttttcccac gagtactgga tgagacatgc cctgaccctg gccaagaggg
     3301 cacgggatga gagggaggtg cctgtgggag ccgtgctggt gctgaacaat agagtgatcg
     3361 gcgagggctg gaacagagcc atcggcctgc acgacccaac agcccatgcc gaaattatgg
     3421 ccctgagaca gggcggcctg gtcatgcaga actacagact gattgacgcc accctgtacg
     3481 tgacattcga gccttgcgtg atgtgcgccg gcgccatgat ccactctagg atcggccgcg
     3541 tggtgtttgg cgtgaggaac tcaaaaagag gcgccgcagg ctccctgatg aacgtgctga
     3601 actaccccgg catgaatcac cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg
     3661 ccgccctgct gtgcgatttc tatcggatgc ctagacaggt gttcaatgct cagaagaagg
     3721 cccagagctc catcaactag tcagggccct attctatagg tgtcacctaa atgctagagc
     3781 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3841 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3901 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3961 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4021 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg ctgcagtgag
     4081 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4141 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4201 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4261 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4321 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4381 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4441 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4501 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4561 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4621 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4681 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4741 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4801 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4861 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4921 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4981 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5041 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5101 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5161 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5221 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5281 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5341 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5401 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5461 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5521 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5581 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5641 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5701 ccccagtgct gcaatgatac cgcgagatcc acgctcaccg gctccagatt tatcagcaat
     5761 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5821 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5881 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5941 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6001 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6061 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6121 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6181 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6241 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6301 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6361 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6421 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6481 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6541 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
