pCAG-mTag-BFP2

LOCUS       pCAG-mTag-BFP2           6629 bp DNA     circular SYN 31-MAR-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6629)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 31, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..39)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    47..63
                     /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(71..101)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    116..137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        196..391
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      242..377
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    397..531
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(770..1358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1529..2389)
                     /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"
     promoter        complement(2390..2494)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    2545..2556
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     enhancer        2560..2939
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2941..3218
                     /label=chicken beta-actin promoter
     intron          3219..4235
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     CDS             4304..5005
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     intron          5041..5173
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    5182..5770
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5653..5664)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5803..5914
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(6010..6028)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6049..6065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6236..6291
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
ORIGIN
        1 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
       61 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
      121 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
      181 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
      241 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
      301 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
      361 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
      421 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
      481 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
      541 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      601 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      661 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      721 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      781 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      841 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      901 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      961 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1021 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1081 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1141 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1201 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1261 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1321 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1381 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1441 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1501 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1561 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1621 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1681 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1741 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1801 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1861 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1921 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1981 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2041 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2101 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2161 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2221 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2281 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2341 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2401 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2461 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2521 ggtcgacatt gattattgac tagagtcatc ctcatcgtcg acattgatta ttgactagtt
     2581 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     2641 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     2701 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     2761 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     2821 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     2881 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     2941 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
     3001 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
     3061 ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg
     3121 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
     3181 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc
     3241 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
     3301 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
     3361 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg
     3421 gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg
     3481 ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt
     3541 gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg
     3601 ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg
     3661 gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc
     3721 acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg
     3781 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg
     3841 gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc
     3901 gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa
     3961 tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga
     4021 agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc
     4081 gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg
     4141 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
     4201 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
     4261 tgctgtctca tcattttggc aaagaattcg accggtcgcc accatgagcg agctgattaa
     4321 ggagaacatg cacatgaagc tgtacatgga gggcaccgtg gacaaccatc acttcaagtg
     4381 cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt
     4441 cgagggcggc cctctcccct tcgccttcga catcctggct actagcttcc tctacggcag
     4501 caagaccttc atcaaccaca cccagggcat ccccgacttc ttcaagcagt ccttccctga
     4561 gggcttcaca tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca
     4621 ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt
     4681 cacatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct tcaccgagac
     4741 gctgtacccc gctgacggcg gcctggaagg cagaaacgac atggccctga agctcgtggg
     4801 cgggagccat ctgatcgcaa acgccaagac cacatataga tccaagaaac ccgctaagaa
     4861 cctcaagatg cctggcgtct actatgtgga ctacagactg gaaagaatca aggaggccaa
     4921 caacgagacc tacgtcgagc agcacgaggt ggcagtggcc agatactgcg acctccctag
     4981 caaactgggg cacaagctta attaagtaca agtaaagcgg ccccggactc gaggccgcag
     5041 gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga
     5101 cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc
     5161 tttctctcca caggtgtcga caatcaacct ctggattaca aaatttgtga aagattgact
     5221 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     5281 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     5341 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     5401 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     5461 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5521 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     5581 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5641 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5701 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5761 gcctccccgc ctggaattcg agctcggtac gatcagcctc gactgtgcct tctagttgcc
     5821 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5881 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     5941 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcccag
     6001 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     6061 tcctgtgtga aattgttatc cgctaattca ctcctcaggt gcaggctgcc tatcagaagg
     6121 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatcga tctttttccc
     6181 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     6241 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     6301 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     6361 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
     6421 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
     6481 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
     6541 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
     6601 cttctcttat ggagatccct cgacctgca
//