LOCUS       sequence-368790-        6450 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 6450)
  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 6450)
  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-UGI
FEATURES             Location/Qualifiers
     source          1..6450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     promoter        complement(1828..1932)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2000..2018
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2056..2078)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2173..2496)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     2391..2413
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2405..2422
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2406..2422
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2423..2479
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2492..2508)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2516..2532
                     /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(2540..2546,2547..2564,2565..2570))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2585..2606
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3360..3611
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     promoter        complement(3624..3642)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3625..3642)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3662..3679)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3668..3892
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4123..4244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4160..4179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4214..4233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4293..4309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4317..4333
                     /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(4341..4347,4348..4365,4366..4371))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4386..4407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4695..5283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4775..4794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5454..6245,6246..6314))
                     /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"
     promoter        complement(6315..6419)
                     /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 gaagagacct ccgtaagatg
     1501 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1561 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1621 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1681 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1741 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     1801 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     1861 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     1921 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     1981 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     2041 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     2101 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     2161 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     2221 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     2281 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     2341 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     2401 acgttgtaaa acgacggcca gtgaattcga gctcggtacc cggggatcct ctagagtcga
     2461 cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2521 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2581 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2641 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2701 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2761 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggggt
     2821 ctcacggtct gactccggaa caggtggtgg cgattgcaag caacggcggc ggcaaacagg
     2881 ctctagagag cattgttgcc cagctctcca gacctgatcc ggcgctagcc gcgttgacaa
     2941 atgatcatct tgttgctctt gcttgtcttg gcggacgacc tgccctggat gctgtgaaaa
     3001 aggggttggg gggctctgga tcaggtagct acgcacttgg tccttaccag attagcgcac
     3061 cccaactccc cgcctataat ggtcaaaccg tcgggacctt ttactacgta aacgatgctg
     3121 gtgggctgga atccaaagta ttctcctcag ggggccctac accctacccc aactacgcca
     3181 atgctggtca tgtagaaggg cagtcagcac tgtttatgcg cgataatggt ataagcgagg
     3241 ggttggtctt ccataacaac ccagagggta cttgtggctt ctgtgtgaat atgactgaaa
     3301 cccttctgcc cgaaaatgcc aagatgactg tcgtcccacc tgaaggctcc ggtggttcaa
     3361 caaatctgtc tgacatcata gaaaaagaaa ctggcaaaca actggtgata caagaaagta
     3421 ttctgatgtt gcctgaggaa gttgaagagg taataggcaa caagcctgaa agcgatatac
     3481 tggtacatac cgcatacgat gagtcaactg acgaaaacgt gatgctcttg acatccgatg
     3541 cccctgaata caagccctgg gcacttgtaa tccaagatag taatggggag aacaaaatca
     3601 aaatgttgta gtcagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     3661 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3721 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3781 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3841 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3901 gcggaaagaa ccagctgggg ctctaggggg tatccccacg ctgcagtgag cgggactctg
     3961 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4021 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4081 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     4141 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4201 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     4261 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4321 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4381 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4441 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4501 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4561 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4621 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4681 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4741 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4801 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4861 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4921 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4981 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5041 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5101 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5161 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5221 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5281 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5341 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5401 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5461 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5521 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5581 gcaatgatac cgcgagatcc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5641 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5701 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5761 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5821 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5881 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5941 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6001 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6061 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6121 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6181 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6241 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6301 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6361 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6421 acatttcccc gaaaagtgcc acctgacgtc
//
