LOCUS       sequence-358832-        4829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express mCerulean3 fused to the N-terminus of the Bcl-2 family 
            chimera protein, Bcl-2-ActA: Bcl-2 with its membrane binding region 
            swapped for ActA (mitochondrial targeting sequence)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4829)
  TITLE     David Andrews Lab plasmids
REFERENCE   2  (bases 1 to 4829)
  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: mCerulean3-Bcl2-ActA-s2193
FEATURES             Location/Qualifiers
     source          1..4829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    100..221
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(111..130)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     165..184
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      409..996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     897..916
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1118..1309
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1141..1161)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     regulatory      2431..2440
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2437..2439,2440..2442,2443..3153)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2482..2503)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2743..2762)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3090..3111
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3921..4365
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        4399..4445
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        4437..4455
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 caagctggtc aaaaccacca ttgaggaact catccccctc aagtacacca ggaactaaac
       61 ctgaattctt cgaactcgag gctagctggc cagacatgat aagatacatt gatgagtttg
      121 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
      181 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
      241 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
      301 acaaatgtgg tatggaaatg ttaattaact agccatgacc aaaatccctt aacgtgagtt
      361 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
      421 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      481 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      541 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
      601 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
      661 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
      721 gggctgaacg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
      781 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
      841 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
      901 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
      961 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     1021 cggttcctgg ccttttgctg gccttttgct cacatgttct taattaacct gcagggcctg
     1081 aaataacctc tgaaagagga acttggttag gtaccttctg aggctgaaag aaccagctgt
     1141 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     1201 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     1261 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtccca ctagttccgc
     1321 cagagcgcgc gagggcctcc agcggccgcc cctcccccac agcaggggcg gggtcccgcg
     1381 cccaccggaa ggagcgggct cggggcgggc ggcgctgatt ggccggggcg ggcctgacgc
     1441 cgacgcggct ataagagacc acaagcgacc cgcagggcca gacgttcttc gccgaagctt
     1501 gccgtcagaa cgcaggtgag gggcgggtgt ggcttccgcg ggccgccgag ctggaggtcc
     1561 tgctccgagc gggccgggcc ccgctgtcgt cggcggggat tagctgcgag cattcccgct
     1621 tcgagttgcg ggcggcgcgg gaggcagagt gcgaggccta gcggcaaccc cgtagcctcg
     1681 cctcgtgtcc ggcttgaggc ctagcgtggt gtccgcgccg ccgccgcgtg ctactccggc
     1741 cgcactctgg tctttttttt ttttgttgtt gttgccctgc tgccttcgat tgccgttcag
     1801 caataggggc taacaaaggg agggtgcggg gcttgctcgc ccggagcccg gagaggtcat
     1861 ggttggggag gaatggaggg acaggagtgg cggctggggc ccgcccgcct tcggagcaca
     1921 tgtccgacgc cacctggatg gggcgaggcc tggggttttt cccgaagcaa ccaggctggg
     1981 gttagcgtgc cgaggccatg tggccccagc acccggcacg atctggcttg gcggcgccgc
     2041 gttgccctgc ctccctaact agggtgaggc catcccgtcc ggcaccagtt gcgtgcgtgg
     2101 aaagatggcc gctcccgggc cctgttgcaa ggagctcaaa atggaggacg cggcagcccg
     2161 gtggagcggg cgggtgagtc acccacacaa aggaagaggg cctggtccct caccggctgc
     2221 tgcttcctgt gaccccgtgg tcctatcggc cgcaatagtc acctcgggct tttgagcacg
     2281 gctagtcgcg gcggggggag gggatgtaat ggcgttggag tttgttcaca tttggtgggt
     2341 ggagactagt caggccagcc tggcgctgga agtcattttt ggaatttgtc cccttgagtt
     2401 ttgagcggag ctaattcaaa gcaaccggtc gccaccatgg tgagcaaggg cgaggagctg
     2461 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2521 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2581 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gagctggggc
     2641 gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2701 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2761 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2821 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa cgccatccac
     2881 ggcaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcggcctc
     2941 aactgcaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     3001 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg
     3061 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     3121 gggatcactc tcggcatgga cgagctgtac aagtccggac tcagatccac gcgtatggcg
     3181 cacgctggga gaacagggta cgataaccgg gagatagtga tgaagtacat ccattataag
     3241 ctgtcgcaga ggggctacga gtgggatgcg ggagatgtgg gcgccgcgcc cccgggggcc
     3301 gcccccgcac cgggcatctt ctcctcccag cccgggcaca cgccccatcc agccgcatcc
     3361 cgggacccgg tcgccaggac ctcgccactg cagaccccgg ctgcccccgg cgccgccgcg
     3421 gggcctgcgc tcagcccggt gccacctgtg gtccacctga ccctccgcca ggccggcgac
     3481 gacttctccc gccgctaccg ccgcgacttc gccgagatgt ccagccagct gcacctgacg
     3541 cccttcaccg cgcggggacg ctttgccacg gtggtggagg agctcttcag ggacggggtg
     3601 aactggggga ggattgtggc cttctttgag ttcggtgggg tcatatgtgt ggagagcgtc
     3661 aaccgggaga tgtcgcccct ggtggacaac atcgccctgt ggatgactga gtacctgaac
     3721 cggcacctgc acacctggat ccaggataac ggaggctggg atgcctttgt ggagctctac
     3781 ggccccagca tgcggcctct gtttgatttc tcctggctgt ctcttaagct gattctcgcc
     3841 atgctggcaa ttggggtgtt ctccctcggc gccttcatta agatcattca gctccggaag
     3901 aacaattagt cgaccctagg agcaggtttc cccaatgaca caaaacgtgc aacttgaaac
     3961 tccgcctggt ctttccaggt ctagaggggt aacactttgt actgcgtttg gctccacgct
     4021 cgatccactg gcgagtgtta gtaacagcac tgttgcttcg tagcggagca tgacggccgt
     4081 gggaactcct ccttggtaac aaggacccac ggggccaaaa gccacgccca cacgggcccg
     4141 tcatgtgtgc aaccccagca cggcgacttt actgcgaaac ccactttaaa gtgacattga
     4201 aactggtacc cacacactgg tgacaggcta aggatgccct tcaggtaccc cgaggtaaca
     4261 cgcgacactc gggatctgag aaggggactg gggcttctat aaaagcgctc ggtttaaaaa
     4321 gcttctatgc ctgaataggt gaccggaggt cggcaccttt cctttgcaat tactgaccct
     4381 atgaatacaa ctgactgttt gacaattaat catcggcata gtatatcggc atagtataat
     4441 acgactcact ataggagggc caccatgaag accttcaaca tctctcagca ggatctggag
     4501 ctggtggagg tcgccactga gaagatcacc atgctctatg aggacaacaa gcaccatgtc
     4561 ggggcggcca tcaggaccaa gactggggag atcatctctg ctgtccacat tgaggcctac
     4621 attggcaggg tcactgtctg tgctgaagcc attgccattg ggtctgctgt gagcaacggg
     4681 cagaaggact ttgacaccat tgtggctgtc aggcacccct actctgatga ggtggacaga
     4741 tccatcaggg tggtcagccc ctgtggcatg tgcagagagc tcatctctga ctatgctcct
     4801 gactgctttg tgctcattga gatgaatgg
//
