LOCUS       sequence-314375-        5397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and linker-mEGFP sequence for C-terminus tagging of 
            human TFAM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5397)
  TITLE     Allen Institute for Cell Sciences Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5397)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: AICSDP-72:TFAM-mEGFP
FEATURES             Location/Qualifiers
     source          1..5397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..37
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     29..46
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     30..46
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1079..1081,1082..1795)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1121..1142)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1382..1401)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1729..1750
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2827..2843)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2827..2843)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2840..2862)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2851..2867
                     /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(2875..2881,2882..2899,2900..2905))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2920..2941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3058..3075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3229..3817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3309..3328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3988..4779,4780..4848))
                     /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     4611..4630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4849..4953)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5021..5039
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5077..5099)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5199..5218
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgacgcgtat
       61 tgggatacct aaagctacag ggaaggaata aaaacagact ttgaagttag tttatgacaa
      121 cacatagatt atacctgaat attaatattt ccgaatcaaa aatatgtacc ttcattcagt
      181 tcatttttgt aaattgtaac agtttgatag tttttgttta gagagaaaag atgttaagaa
      241 ttgatgaggc tttgcctgta attcttggta tcttgagttt ttgttggaat gaataataaa
      301 ggtcactgaa gtccccatat ctaccttaac ttaaacatat attgttttac aggaaaagct
      361 gaagactgta aaggaaaact ggaaaaatct gtctgactct gaaaaggaag tgagtattac
      421 ggttgttagt ctcaagtgtc tacttaggat atctgtgaga gaatgtatta gggcaaggct
      481 tgattcatgt gaagacaacc ttgtattact aataatacaa gtattctaaa attattttca
      541 tattctcaga accagtttgg catttttctc ttggtagttt ttgtctgcaa ataaaattgg
      601 tatttgaaac cacagcactc ataagcctga gtgtccactt attcagtggc agtgataccc
      661 tgctgctgcc cccttcctgc acctcaagat ctgaacttgt aacaatttca ttcaaagaga
      721 taacagtcta tctcttacat tttcatcatc taacttttat ctcatgaaaa tatttcttct
      781 ccagtctgcc tttatacatg tagaatggta atgtaatttc taaagtatat acagaagcat
      841 tccagaaatt aattgctatt ttaaataatc attttatctc aaaaaataag taaatcattt
      901 taacagttat gctttttctc agttatatat tcagcatgct aaagaggacg aaactcgtta
      961 tcataatgaa atgaagtctt gggaagaaca aatgattgaa gttggacgaa aggatcttct
     1021 acgtcgcaca ataaagaaac aacgaaaata tggtgctgaa gagtgtggag gaggcagcgt
     1081 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1141 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1201 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1261 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1321 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1381 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1441 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1501 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1561 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1621 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1681 gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1741 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaaagta
     1801 gaagattgag atgtgttcac aatggatagc cacaggaaac gagttaggtc tcaatacctg
     1861 aagctatcgt aaaattaaga aaggataaag ttggtaaacc ttttatattt agtatctttt
     1921 tattcagctc atggacttct gccagcataa tacttgcttt ggaaaaccca gataaaggtt
     1981 catgcaaact ttattttgtg tttaggaact actgaggatc agagtaatcc aagcaaatgt
     2041 gaatcatttt acctttgaca aaggtaaatc agactatgaa gtttttttta tacaggatga
     2101 tgactatgga aagagtactc ttgtttcctt atattatgga ggcaggagtt tcgttttcaa
     2161 aattgttaca aattgtagaa gccacggtgt tctgtgatat aagtgtgtgt ttttcataaa
     2221 gcaggcagaa ctcatctagg taaattacag ttcctaggta taattcacat tgtattcaga
     2281 gttgatggtt gtacatataa gtgattgctg gttttagttg caactttgta taaaagggac
     2341 tgagaaattt ataaactttt ttcttactgt cttttttcta aagtaaaaac aaagaaatta
     2401 tgtgccagat ttatgcatat tattttatgt tgcatagaat aaaattttta atctttaatt
     2461 ttacatttcc taaatatatt ttaagacgaa acatttgttc tatagctttt cccttttttt
     2521 aagtaaggaa ttttattttt ttctgaatta ttttctctcg tgagtatatt gatccagaaa
     2581 gaaaacttgt attatgtgtg ttttaaaatg agaaatctaa aaaacgaaaa gtctccaaag
     2641 tctctggaat ttgaaacact ttgcataacg tataaaagcc tgtttaagag acagccaact
     2701 atggcctgtg gatcaaatcc agcctgctgc ctgcttttta tggcctgtga gctaggaatt
     2761 gtgtttataa ttttaaatgt ttttttttaa agatcccaat ggcgcgccga gcttggcgta
     2821 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2881 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2941 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3001 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3061 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3121 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3181 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3241 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3301 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3361 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3421 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3481 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3541 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3601 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3661 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3721 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3781 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3841 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3901 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3961 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4021 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4081 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4141 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     4201 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4261 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4321 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4381 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4441 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4501 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4561 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4621 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4681 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4741 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4801 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4861 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4921 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4981 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     5041 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     5101 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     5161 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     5221 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     5281 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     5341 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggt
//
