LOCUS       sequence-76195-e        5810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a GFP-dependent transcription factor component in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5810)
  AUTHORS   Tang JC, Szikra T, Kozorovitskiy Y, Teixiera M, Sabatini BL, Roska 
            B, Cepko CL
  TITLE     A nanobody-based system using fluorescent proteins as scaffolds for 
            cell-specific gene manipulation.
  JOURNAL   Cell. 2013 Aug 15;154(4):928-39. doi: 10.1016/j.cell.2013.07.021.
  PUBMED    23953120
REFERENCE   2  (bases 1 to 5810)
  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"; doi: 
            "10.1016/j.cell.2013.07"; journalName: "Cell"; date: "2013-08-15- 
            15"; volume: "154"; issue: "4"; pages: "928-39"
COMMENT     Sequence Label: pCAG-rTetRDBD-GBP1
FEATURES             Location/Qualifiers
     source          1..5810
                     /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"
     CDS             2404..2745
                     /codon_start=1
                     /product="GFP-binding fragment of a single-chain camelid 
                     antibody (Rothbauer et al., 2008)"
                     /label=GFP nanobody
                     /translation="VQLVESGGALVQPGGSLRLSCAASGFPVNRYSMRWYRQAPGKERE
                     WVAGMSSAGDRSSYEDSVKGRFTISRDDARNTVYLQMNSLKPEDTAVYYSNVNVGFEYW
                     GQGTQVTVSS"
     primer_bind     complement(2834..2853)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2899..2954
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2953..2972)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3313..3329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3313..3329)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3326..3348)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3337..3353
                     /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(3361..3367,3368..3385,3386..3391))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3406..3427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3486..3681
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3532..3667
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3687..3821
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3724..3743)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3778..3797
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3889..3906)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4060..4648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4140..4159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4819..5610,5611..5679))
                     /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     5442..5461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5680..5784)
                     /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 tgccaccatg agccggctgg acaagagcaa agtgatcaac
     1801 agcgccctgg aactgctgaa cggcgtgggc atcgagggcc tgaccacccg gaagctggcc
     1861 cagaaactgg gcgtggaaca gcccaccctg tactggcacg tgaagaacaa gcgggccctg
     1921 ctggacgccc tgcccatcga gatgctggac cggcaccaca cccacagctg ccctctggaa
     1981 ggcgagagct ggcaggactt cctgcggaac aacgccaaga gctacagatg cgccctgctg
     2041 agccaccggg acggcgccaa agtgcacctg ggcaccagac ccaccgagaa gcagtacgag
     2101 acactggaaa accagctggc cttcctgtgc cagcagggct tcagcctgga aaacgccctg
     2161 tacgccctga gcgccgtggg ccacttcacc ctgggctgcg tgctggaaga acaggaacac
     2221 caggtcgcca aagaggaaag agagacaccc accaccgaca gcatgccccc cctgctgaag
     2281 caggccatcg agctgttcga cagacagggc gccgagcccg ccttcctgtt cggcctggaa
     2341 ctgatcatct gcggcctcga gaagcagctg aagtgcgaga gcggcggagc tagcatggcc
     2401 gacgtgcagc tcgtggaatc cggaggagca ctggtccagc ctgggggatc actgagactg
     2461 tcttgtgccg cttctggctt tcctgtgaat cggtactcta tgagatggta caggcaggca
     2521 cctggaaaag agcgagaatg ggtcgccgga atgtcatctg ctggcgatag gagttcctac
     2581 gaggatagcg tgaaaggacg gttcaccatt agtagggacg atgctagaaa caccgtgtac
     2641 ctccagatga actccctgaa acctgaggac actgctgtgt actactccaa cgtcaatgtg
     2701 ggctttgaat actggggaca gggcacacaa gtgactgtta gttcttaagc ggccgcactc
     2761 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     2821 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     2881 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     2941 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     3001 agtatttggt ttagagtttg gcaacatatg ccatatgctg gctgccatga acaaaggtgg
     3061 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     3121 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     3181 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     3241 ctcccagtca tagctgtccc tcttctctta tgaagatccc tcgacctgca gcccaagctt
     3301 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3361 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3421 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg
     3481 gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3541 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3601 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3661 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat
     3721 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3781 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg
     3841 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3901 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3961 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     4261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4501 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4741 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4801 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4861 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4921 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4981 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5041 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5101 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5161 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5221 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5281 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5341 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5401 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5461 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5521 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5581 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5641 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5701 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5761 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
