LOCUS       sequence-160076-        5891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP-dependent dGBP1-TagBFP, for expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5891)
  AUTHORS   Tang JC, Drokhlyansky E, Etemad B, Rudolph S, Guo B, Wang S, Ellis 
            EG, Li JZ, Cepko CL
  TITLE     Detection and manipulation of live antigen-expressing cells using 
            conditionally stable nanobodies.
  JOURNAL   Elife. 2016 May 20;5. pii: e15312. doi: 10.7554/eLife.15312.
  PUBMED    27205882
REFERENCE   2  (bases 1 to 5891)
  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: "Elife."; 
            date: "2016-05-20"; volume: "5. pii: e15312. doi"; pages: " 
            10.7554/eLife.15312"
COMMENT     Sequence Label: pCAG-dGBP1-TagBFP
FEATURES             Location/Qualifiers
     source          1..5891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1763..1772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2126..2827
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(2913..2932)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2978..3033
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3032..3051)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3394..3410)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3394..3410)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3407..3429)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3418..3434
                     /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(3442..3448,3449..3466,3467..3472))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3487..3508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3567..3762
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3613..3748
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3768..3902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3805..3824)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3859..3878
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3970..3987)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4141..4729)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4221..4240)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4900..5691,5692..5760))
                     /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     5523..5542
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5761..5865)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attctgcagt cgacggtacc
     1741 gcgggcccgg gatccaccgg tcgccaccat ggccgacgtg cagctcgtgg aatccggagg
     1801 agcactggtc cagcctgggg gatcactgag actgtcttgt gtcgcttctg gctttcctgt
     1861 gaatcggtac tctatgagat ggtacaggca ggcacctgga aaagagcgag aatgggtcgc
     1921 cggaatgtca tctgctggcg ataggagttc ctacgtggat agcgtgaaag gacggttcac
     1981 cattagaagg gacgatgcta gaaacaccgt gtacctccag atgaactccc tgaaacctga
     2041 ggacactgct gtgtactact acaacgtcaa tgtgggcttt gaatactggg gacatggcac
     2101 acaagtgact gttagttttg catgcatgag cgagctgatt aaggagaaca tgcacatgaa
     2161 gctgtacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg
     2221 caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc
     2281 cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca
     2341 cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca catgggagag
     2401 agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga
     2461 cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt
     2521 gatgcagaag aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg
     2581 cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc
     2641 aaacatcaag accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt
     2701 ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga
     2761 gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct
     2821 taattaagcg gccgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     2881 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     2941 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     3001 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     3061 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     3121 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     3181 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     3241 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     3301 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     3361 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3421 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3481 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3541 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     3601 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3661 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3721 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     3781 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3841 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     3901 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3961 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4021 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4081 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4141 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4201 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4261 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4321 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4381 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4441 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4501 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4561 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4621 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4681 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4741 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4801 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4861 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4921 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4981 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5041 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5101 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5161 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5221 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5281 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5341 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5401 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5461 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5521 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5581 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5641 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5701 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5761 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5821 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5881 agtgccacct g
//
