LOCUS       sequence-347947-        4607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express mCerulean3 fused to the N-terminus of the Bcl-2 family 
            protein, Bik, in a plasmid with Blasticidin resistance. Used to make
            stable BMK-DKO cell lines..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4607)
  TITLE     David Andrews Lab plasmids
REFERENCE   2  (bases 1 to 4607)
  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-Bik-s2193
FEATURES             Location/Qualifiers
     source          1..4607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    33..154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(44..63)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     98..117
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      342..929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     830..849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1051..1242
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1074..1094)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     regulatory      2364..2373
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2370..2372,2373..2375,2376..3086)
                     /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(2415..2436)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2676..2695)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3023..3044
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3632..4076
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        4110..4156
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        4148..4166
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cttcgaactc gaggctagct ggccagacat gataagatac attgatgagt ttggacaaac
       61 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
      121 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
      181 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
      241 tggtatggaa atgttaatta actagccatg accaaaatcc cttaacgtga gttttcgttc
      301 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      361 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      421 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      481 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      541 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      601 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      661 acgggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
      721 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
      781 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
      841 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
      901 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
      961 tggccttttg ctggcctttt gctcacatgt tcttaattaa cctgcagggc ctgaaataac
     1021 ctctgaaaga ggaacttggt taggtacctt ctgaggctga aagaaccagc tgtggaatgt
     1081 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     1141 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     1201 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccactagttc cgccagagcg
     1261 cgcgagggcc tccagcggcc gcccctcccc cacagcaggg gcggggtccc gcgcccaccg
     1321 gaaggagcgg gctcggggcg ggcggcgctg attggccggg gcgggcctga cgccgacgcg
     1381 gctataagag accacaagcg acccgcaggg ccagacgttc ttcgccgaag cttgccgtca
     1441 gaacgcaggt gaggggcggg tgtggcttcc gcgggccgcc gagctggagg tcctgctccg
     1501 agcgggccgg gccccgctgt cgtcggcggg gattagctgc gagcattccc gcttcgagtt
     1561 gcgggcggcg cgggaggcag agtgcgaggc ctagcggcaa ccccgtagcc tcgcctcgtg
     1621 tccggcttga ggcctagcgt ggtgtccgcg ccgccgccgc gtgctactcc ggccgcactc
     1681 tggtcttttt tttttttgtt gttgttgccc tgctgccttc gattgccgtt cagcaatagg
     1741 ggctaacaaa gggagggtgc ggggcttgct cgcccggagc ccggagaggt catggttggg
     1801 gaggaatgga gggacaggag tggcggctgg ggcccgcccg ccttcggagc acatgtccga
     1861 cgccacctgg atggggcgag gcctggggtt tttcccgaag caaccaggct ggggttagcg
     1921 tgccgaggcc atgtggcccc agcacccggc acgatctggc ttggcggcgc cgcgttgccc
     1981 tgcctcccta actagggtga ggccatcccg tccggcacca gttgcgtgcg tggaaagatg
     2041 gccgctcccg ggccctgttg caaggagctc aaaatggagg acgcggcagc ccggtggagc
     2101 gggcgggtga gtcacccaca caaaggaaga gggcctggtc cctcaccggc tgctgcttcc
     2161 tgtgaccccg tggtcctatc ggccgcaata gtcacctcgg gcttttgagc acggctagtc
     2221 gcggcggggg gaggggatgt aatggcgttg gagtttgttc acatttggtg ggtggagact
     2281 agtcaggcca gcctggcgct ggaagtcatt tttggaattt gtccccttga gttttgagcg
     2341 gagctaattc aaagcaaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     2401 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2461 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2521 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgagctgg ggcgtgcagt
     2581 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2641 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2701 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2761 tcaaggagga cggcaacatc ctggggcaca agctggagta caacgccatc cacggcaacg
     2821 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcggc ctcaactgca
     2881 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2941 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag
     3001 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3061 ctctcggcat ggacgagctg tacaagtctg gactcagatc ccgaggaggc atgtctgaag
     3121 taagacccct ctccagagac atcttgatgg agaccctcct gtatgagcag ctcctggaac
     3181 ccccgaccat ggaggttctt ggcatgactg actctgaaga ggacctggac cctatggagg
     3241 acttcgattc tttggaatgc atggagggca gtgacgcatt ggccctgcgg ctggcctgca
     3301 tcggggacga gatggacgtg agcctcaggg ccccgcgcct ggcccagctc tccgaggtgg
     3361 ccatgcacag cctgggtctg gctttcatct acgaccagac tgaggacatc agggatgttc
     3421 ttagaagttt catggacggt ttcaccacac ttaaggagaa cataatgagg ttctggagat
     3481 ccccgaaccc cgggtcctgg gtgtcctgcg aacaggtgct gctggcgctg ctgctgctgc
     3541 tggcgctgct gctgccgctg ctcagcgggg gcctgcacct gctgctcaag tgactcgagc
     3601 tcaagcttcg aattctgcag tcgaccctag gagcaggttt ccccaatgac acaaaacgtg
     3661 caacttgaaa ctccgcctgg tctttccagg tctagagggg taacactttg tactgcgttt
     3721 ggctccacgc tcgatccact ggcgagtgtt agtaacagca ctgttgcttc gtagcggagc
     3781 atgacggccg tgggaactcc tccttggtaa caaggaccca cggggccaaa agccacgccc
     3841 acacgggccc gtcatgtgtg caaccccagc acggcgactt tactgcgaaa cccactttaa
     3901 agtgacattg aaactggtac ccacacactg gtgacaggct aaggatgccc ttcaggtacc
     3961 ccgaggtaac acgcgacact cgggatctga gaaggggact ggggcttcta taaaagcgct
     4021 cggtttaaaa agcttctatg cctgaatagg tgaccggagg tcggcacctt tcctttgcaa
     4081 ttactgaccc tatgaataca actgactgtt tgacaattaa tcatcggcat agtatatcgg
     4141 catagtataa tacgactcac tataggaggg ccaccatgaa gaccttcaac atctctcagc
     4201 aggatctgga gctggtggag gtcgccactg agaagatcac catgctctat gaggacaaca
     4261 agcaccatgt cggggcggcc atcaggacca agactgggga gatcatctct gctgtccaca
     4321 ttgaggccta cattggcagg gtcactgtct gtgctgaagc cattgccatt gggtctgctg
     4381 tgagcaacgg gcagaaggac tttgacacca ttgtggctgt caggcacccc tactctgatg
     4441 aggtggacag atccatcagg gtggtcagcc cctgtggcat gtgcagagag ctcatctctg
     4501 actatgctcc tgactgcttt gtgctcattg agatgaatgg caagctggtc aaaaccacca
     4561 ttgaggaact catccccctc aagtacacca ggaactaaac ctgaatt
//
