LOCUS       sequence-368783-        6294 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 6294)
  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 6294)
  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_Left-stuffer-1397N
FEATURES             Location/Qualifiers
     source          1..6294
                     /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             978..1004
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1008..1034
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1038..1064
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1490..2164
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1490..2164
                     /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(1620..1638)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1888..1907
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2082..2105
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             join(2206..2208,2209..2922)
                     /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(2206..2208,2209..2919)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2248..2269)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2509..2528)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2856..2877
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3468..3486)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3469..3486)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3506..3523)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3512..3736
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    3967..4088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4004..4023)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4058..4077
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4137..4153)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4137..4153)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4150..4172)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4161..4177
                     /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(4185..4191,4192..4209,4210..4215))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4230..4251
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4368..4385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4539..5127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4619..4638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5298..6089,6090..6158))
                     /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     5921..5940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6159..6263)
                     /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 atgggtggag 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 ggagatggct cttagtcgag
      901 ccgtctgtgg tacttcccga cagcttgctc cagtcttggg gtaccttggc tcccgacaaa
      961 agcactctct ccctgattac ccttacgatg tacctgacta cgccggctac ccttacgacg
     1021 taccagatta cgctggatat ccatacgatg tccctgatta tgctgggatc cgaattcaag
     1081 atctacgcac gctcggctac agccagcagc aacaggagaa gatcaaaccg aaggttcgtt
     1141 cgacagtggc gcagcaccac gaggcactgg tcggccatgg gtttacacac gcgcacatcg
     1201 ttgcgctcag ccaacacccg gcagcgttag ggaccgtcgc tgtcaagtat caggacatga
     1261 tcgcagcgtt gccagaggcg acacacgaag cgatcgttgg cgtcggcaaa cagtggtccg
     1321 gcgcacgcgc tctggaggcc ttgctcacgg tggcgggaga gttgagaggt ccaccgttac
     1381 agttggacac aggccaactt ctcaagattg caaaacgtgg cggcgtgacc gcagtggagg
     1441 cagtgcatgc atggcgcaat gcactgacgg gtgcccccct gaggagacga tggcctcctc
     1501 cgaggacgtc atcaaggagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg
     1561 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     1621 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccctca
     1681 gttccagtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     1741 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     1801 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     1861 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     1921 ggcctccacc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagatgag
     1981 gctgaagctg aaggacggcg gccactacga cgccgaggtc aagaccacct acatggccaa
     2041 gaagcccgtg cagctgcccg gcgcctacaa gaccgacatc aagctggaca tcacctccca
     2101 caacgaggac tacaccatcg tggaacagta cgagcgcgcc gagggccgcc actccaccgg
     2161 cgccgaattc atgacatcaa ttattataca tcggaggagg atccagtgag caagggcgag
     2221 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2281 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2341 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     2401 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     2461 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     2521 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2581 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2641 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     2701 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     2761 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     2821 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2881 gccgccggga tcactctcgg catggacgag ctgtacaagt aacgtctccc ggtctgactc
     2941 cggaacaggt ggtggcgatt gcaagcaacg gcggcggcaa acaggctcta gagagcattg
     3001 ttgcccagct ctccagacct gatccggcgc tagccgcgtt gacaaatgat catcttgttg
     3061 ctcttgcttg tcttggcgga cgacctgccc tggatgctgt gaaaaagggg ttggggggct
     3121 ctggatcagg tagctacgca cttggtcctt accagattag cgcaccccaa ctccccgcct
     3181 ataatggtca aaccgtcggg accttttact acgtaaacga tgctggtggg ctggaatcca
     3241 aagtattctc ctcagggggc cctacaccct accccaacta cgccaatgct ggtcatgtag
     3301 aagggcagtc agcactgttt atgcgcgata atggtataag cgaggggttg gtcttccata
     3361 acaacccaga gggtacttgt ggcttctgtg tgaatatgac tgaaaccctt ctgcccgaaa
     3421 atgccaagat gactgtcgtc ccacctgaag gctagtcagg gccctattct atagtgtcac
     3481 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3541 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3601 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3661 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3721 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3781 cacgctgcag tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     3841 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     3901 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     3961 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4021 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4081 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4141 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4201 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4261 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4321 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4381 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4441 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4501 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4561 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4621 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4681 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4741 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4801 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4861 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4921 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4981 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5041 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5101 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5161 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5221 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5281 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5341 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5401 ggcttaccat ctggccccag tgctgcaatg ataccgcgag atccacgctc accggctcca
     5461 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5521 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5581 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5641 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5701 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5761 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5821 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5881 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5941 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6001 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6061 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6121 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6181 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6241 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
