LOCUS       sequence-479362-        3052 bp    DNA     circular SYN 22-JUL-2025
DEFINITION  Plasmids for constructing mitoBEs v2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3052)
  AUTHORS   Zhang X, Zhang X, Ren J, Li J, Wei X, Yu Y, Yi Z, Wei W
  TITLE     Precise modelling of mitochondrial diseases using optimized mitoBEs.
  JOURNAL   Nature. 2025 Jan 22. doi: 10.1038/s41586-024-08469-8.
  PUBMED    39843744
REFERENCE   2  (bases 1 to 3052)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2025 Jan 22. doi: 10.1038/s41586-024-08469-8."
COMMENT     Sequence Label: pCMV-COX8 MTS-3xFlag-TALE
            array-CBE6d-UGI-P2A-...
FEATURES             Location/Qualifiers
     source          1..3052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(53..157)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(236..255)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        427..806
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(1083..1099)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1083..1099)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1096..1118)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1107..1123
                     /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(1131..1137,1138..1155,1156..1161))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1176..1197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1314..1331)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1485..2073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1565..1584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2244..3035,3036..3052,1..52))
                     /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     2867..2886
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
       61 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      121 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      181 ctgacgtcga cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa
      241 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      301 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      361 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      421 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      481 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      541 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      601 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      661 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      721 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      781 acgtattagt catcgctatt accatggtga tgcggttttg gcagtactgt ctctctcctt
      841 gcccctaacc caaaaagctt catttttctg tgtaggctgc acaagagcct tgattgaaga
      901 tatattcttt ctgaacagta tttaaggttt ccaataaaat gtacacccct cagaaaccgg
      961 tgtttaaacg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     1021 gcggaaagaa ccagctgggg ctcgataccg tcgacctcta gctagagctt ggcgtaatca
     1081 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1141 gccggaagca taaagtgtaa agcctagggt gcctaatgag tgagctaact cacattaatt
     1201 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1261 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1321 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1381 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1441 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1501 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1561 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1621 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1681 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1741 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1801 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1861 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1921 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1981 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2041 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2101 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2161 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2221 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2281 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2341 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2401 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2461 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2521 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2581 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2641 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2701 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2761 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2821 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     2881 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2941 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3001 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gc
//
