LOCUS       sequence-427395-        5387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  targets ECFP-RspANFAST to mitochondrial membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5387)
  TITLE     Tebo lab plasmids 2024
REFERENCE   2  (bases 1 to 5387)
  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"
COMMENT     Sequence Label: pCMV-tomm20-ECFP-RspANFAST
FEATURES             Location/Qualifiers
     source          1..5387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(133..135,136..138,139..849)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(178..199)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(439..458)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     786..807
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1256..1377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1267..1286)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1321..1340
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1492..1538
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1568..1704
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1749..1820
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2046..2634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2126..2145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3380..3448,3449..4240)
                     /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     complement(3598..3617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4253..4389
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4541..4612
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4617..4663)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4681..4729
                     /note="synthetic polyadenylation signal"
     enhancer        4810..5113
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5115..5317
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5268..5288
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atggtgggtc ggaacagcgc catcgccgcg ggcgtgtgcg gtgccctctt catagggtac
       61 tgcatctact ttgaccgcaa aagacgaagt gaccccaact tcggatccag tgctggtggt
      121 agtgctggtg gtatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      181 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      241 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      301 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc
      361 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      421 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      481 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      541 atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac
      601 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc
      661 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      721 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
      781 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
      841 ctgtacaagg gagcaagtgg aatggagacc gtgagattcg gcggcgacga catcgagaac
      901 agcctggcca agatggacga caaggccctg gacaagctgg ccttcggcgc catccagctg
      961 gacggcaacg gcaagatcat ccactacaac gccgccgagg gcaccatcac cggcagagac
     1021 cccaagaccg tgatcggcaa gaacttcttc accgacgtgg cccccggcac ccagagcaag
     1081 gagttccagg gcagattcaa ggagggcgtg cagaagggcg acctgaacac catgttcgag
     1141 tggatgatcc ccaccagcag aggccccacc aaggtgaagg tgcacatgaa gaaggccatg
     1201 acctaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga
     1261 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1321 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1381 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     1441 agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac
     1501 cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta
     1561 tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     1621 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     1681 tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc
     1741 gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat
     1801 caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc
     1861 agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc
     1921 gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt
     1981 taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2041 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2101 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2161 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2221 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2281 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2341 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2401 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2461 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2521 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2581 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2641 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2701 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2761 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2821 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
     2881 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
     2941 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
     3001 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
     3061 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
     3121 tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     3181 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     3241 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     3301 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     3361 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     3421 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3481 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3541 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     3601 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3661 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     3721 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3781 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3841 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3901 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     3961 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     4021 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4081 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4141 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4201 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     4261 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     4321 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     4381 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     4441 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     4501 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     4561 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     4621 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     4681 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     4741 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     4801 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     4861 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     4921 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     4981 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     5041 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     5101 cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     5161 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     5221 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     5281 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag
     5341 atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
