LOCUS       sequence-368784-        6216 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 6216)
  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 6216)
  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-1397C-UGI
FEATURES             Location/Qualifiers
     source          1..6216
                     /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             3126..3377
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     promoter        complement(3390..3408)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3391..3408)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3428..3445)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3434..3658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    3889..4010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3926..3945)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3980..3999
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4059..4075)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4083..4099
                     /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(4107..4113,4114..4131,4132..4137))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4152..4173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4461..5049)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4541..4560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5220..6011,6012..6080))
                     /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(6081..6185)
                     /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 gggctctggc agtgccatac ctgtgaagcg gggagcaaca ggggagacaa
     3061 aggtgttcac aggcaactct aacagtccaa agagccccac caaaggcggg tgttccggtg
     3121 gttcaacaaa tctgtctgac atcatagaaa aagaaactgg caaacaactg gtgatacaag
     3181 aaagtattct gatgttgcct gaggaagttg aagaggtaat aggcaacaag cctgaaagcg
     3241 atatactggt acataccgca tacgatgagt caactgacga aaacgtgatg ctcttgacat
     3301 ccgatgcccc tgaatacaag ccctgggcac ttgtaatcca agatagtaat ggggagaaca
     3361 aaatcaaaat gttgtagtca gggccctatt ctatagtgtc acctaaatgc tagagctcgc
     3421 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3481 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3541 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3601 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3661 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgctgc agtgagcggg
     3721 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3781 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3841 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     3901 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3961 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4021 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4081 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4141 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4201 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4261 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4321 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4381 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4441 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4501 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4561 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4621 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4681 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4741 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4801 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4861 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4921 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4981 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5041 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5101 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5161 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5221 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5281 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5341 agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc agcaataaac
     5401 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5461 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5521 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5581 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5641 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5701 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5761 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5821 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5881 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5941 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6001 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6061 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6121 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6181 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
