LOCUS       sequence-404777-        6139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Combine with Gal4 to direct CaMBI 300 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6139)
  AUTHORS   Porta-de-la-Riva M, Gonzalez AC, Sanfeliu-Cerdan N, Karimi S, 
            Malaiwong N, Pidde A, Morales-Curiel LF, Fernandez P, 
            Gonzalez-Bolivar S, Hurth C, Krieg M
  TITLE     Neural engineering with photons as synaptic transmitters.
  JOURNAL   Nat Methods. 2023 Apr 6. doi: 10.1038/s41592-023-01836-9.
  PUBMED    37024651
REFERENCE   2  (bases 1 to 6139)
  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: "Nat 
            Methods. 2023 Apr 6. doi: 10.1038/s41592-023-01836-9."
COMMENT     Sequence Label: pNMSB109
FEATURES             Location/Qualifiers
     source          1..6139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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)."
     primer_bind     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    244..338
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    377..471
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    510..604
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     CDS             2622..2699
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     promoter        4092..4196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4197..4265,4266..5057)
                     /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     complement(4415..4434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5228..5816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5717..5736
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5970..5987
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6104..6125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgc caagctgtaa gtttaaacat gatcttacta
      121 actaactatt ctcatttaaa ttttcagagc ttaaaaatgg ctgaaatcac tcacaacgat
      181 ggatacgcta acaacttgga aatgaaataa gcttgctagc ccatgattac gccaagctac
      241 gggcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact gtcctccgag
      301 cggagtactg tcctccgagc ggagtactgt cctccgagcg gagactccct agcccatgat
      361 tacgccaagc tacgggcgga gtactgtcct ccgagcggag tactgtcctc cgagcggagt
      421 actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagactc
      481 cctagcccat gattacgcca agctacgggc ggagtactgt cctccgagcg gagtactgtc
      541 ctccgagcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc
      601 cgagcggaga ctccctaggg gcggccgcac tgactgggcc ggccaaaaat gcataaggtt
      661 ttgctggcac tgttctttat ctttctggca ccagcatccg cactggcagt ctccgaaccg
      721 gcctgcaggt cgattttttg caaattacga gcgttgtagg gggcggagcg ataggtccta
      781 taggttttgg tatatcatca ttcattcatt cattggtaca ttcatttacc caccttcctc
      841 tttctgagct tctctggagt tctgtgcttc ctttttccct tatctttata ctgtaatttt
      901 taactttcag gcattgattg gcgcgcctct agaggatccc cgggattggc caaaggaccc
      961 aaaggtatgt ttcgaatgat actaacataa catagaacat tttcaggagg acccttggct
     1021 agcgtcgacg gtaccggtag atggtttcta agggagaaga acttatcaag gagaacatgc
     1081 gctctaagct ttaccttgag ggatctgtta acggacatca attcaagtgc actcacgagg
     1141 gagaaggaaa gccatatgaa ggaaagcaaa ctaaccgcat caaggttgtt gaaggaggac
     1201 cacttccatt cgctttcgac atccttgcta ctcatttcat gtacggatct aaggttttca
     1261 ttaagtaccc agctgatctt ccagactact tcaagcaatc tttcccagaa ggattcactt
     1321 gggagcgtgt tatggttttc gaggatggag gagtccttac cgctacccaa gatacctccc
     1381 ttcaagatgg agagcttatc tacaacgtta aggttcgtgg agttaatttc ccagctaacg
     1441 gaccagttat gcaaaagaag actcttggat gggagccatc tactgagacc atgtacccag
     1501 ccgatggagg acttgaggga cgctgcgaca aggctcttaa gcttgttgga ggaggacacc
     1561 ttcacgttaa cttcaagact acttacaagt ctaaaaagcc agttaagatg ccaggagttc
     1621 attacgtcga tcgtcgcctt gagcgtatca aggaggctga taacgagact tacgttgaac
     1681 aatacgagca cgccgtcgct cgttactcta accttggagg aggattcact cttgaggact
     1741 tcgtcggaga ttggcgccaa accgctggat acaaccttga tcaagttctc gagcaaggag
     1801 gagtttcttc ccttttccaa aaccttggag tttctgttac cccaatccaa cgcatcgttc
     1861 tttctggaga gaacggactt aaggtaagtt taaacatata tatactaact aaccctgatt
     1921 atttaaattt tcagatcgat atccacgtta tcatcccata cgagggactt tctggagatc
     1981 agatgggaca aatcgagaag atcttcaagg ttgtttaccc agtcgacgac caccacttca
     2041 aggttatcct tcactacgga acccttgtta tcgatggagt cactccaaac atgattgatt
     2101 acttcggacg cccatacgag ggaatcgctg ttttcgatgg aaagaagatc accgttactg
     2161 gaacccttat catgcacgat caacttactg aggaacaaat cgctgaattc aaggaggctt
     2221 tctccctttt cgataaggac ggagatggaa ctatcactac caaggagctc ggaaccgtca
     2281 tgcgttctct cggacaaaac ccaactgagg ctgagcttca agacatgatc aacgaggttg
     2341 acgccgacgg aaacggaact atctacttcc cagagttcct caccatgatg gcccgcaaga
     2401 tgaaggatac cgattctgag gaggagatcc gcgaggcttt ccgtgtcttc gataaggacg
     2461 gaaacggata catttccgcc gcccaacttc gtcacgttat gactaacctt ggagagaagc
     2521 ttaccgacga ggaggtcgac gagatgatcc gtgaggctga catcgacgga gacggacaag
     2581 ttaactacga ggagttcgtt caaatgatga ctgctaaggg aaagcgccgt tggaagaaga
     2641 acttcatcgc tgtctccgcc gctaaccgct tcaagaagat ctcttcttcc ggagcccttg
     2701 aactttggaa cggaaacaag gtaagtttaa acagttcggt actaactaac catacatatt
     2761 taaattttca gatcattgat gagcgtctta tcaacccaga tggatccctt cttttccgcg
     2821 ttactatcaa cggagttact ggatggcgcc tttgcgagcg tatccttgct cgccacgagc
     2881 ttatcaagga gaacatgcgc tctaagcttt accttgaggg atctgttaac ggacaccagt
     2941 tcaagtgcac ccacgaagga gagggaaagc catatgaggg aaagcaaact aaccgcatca
     3001 aggttgttga gggaggacca cttccattcg ctttcgacat ccttgccact cacttcatgt
     3061 acggatccaa ggttttcatc aagtacccag ctgaccttcc agattacttc aagcaatctt
     3121 tcccagaggg attcacctgg gagcgtgtta tggttttcga ggacggagga gtccttaccg
     3181 ccacccaaga cacctccctc caagacggag agcttatcta caacgttaag gttcgtggag
     3241 ttaacttccc agctaacgga ccagttatgc agaagaagac tcttggatgg gagccatcca
     3301 ctgagaccat gtacccagct gacggaggac ttgagggacg ctgcgacaag gccctcaagc
     3361 ttgttggagg aggacacctt cacgttaact tcaagaccac ttacaagtct aagaaaccag
     3421 ttaagatgcc aggagttcac tacgtcgacc gccgccttga gcgtatcaag gaggccgaca
     3481 acgagactta cgttgagcaa tacgagcacg ctgtcgctcg ttactctaac cttggaggag
     3541 gaatggacga gctttacaag taaggtaccg aattcgctag ccggccatac aagtaatccg
     3601 gatgatcgac gccaacgtcg ttgaattttc aaattttaaa tactgaatat ttgttttttt
     3661 tcctattatt tatttattct ctttgtgttt tttttcttgc tttctaaaaa attaattcaa
     3721 tccaaatcta aacatttttt tttctctttc cgtctcccaa ttcgtattcc gctcctctca
     3781 tctgaacaca atgtgcaagt ttatttatct tctcgctttc atttcattag gacgtggggg
     3841 gaattggtgg aagggggaaa cacacaaaag gatgatggaa atgaaataag gacacacaat
     3901 atgcaacaac attcaattca gaaatatgga ggaaggttta aaagaaaaca taaaaatata
     3961 tagaggagga aggaaaacta gtaaaaaata agcaaagaaa ttaggcgaac gatgagaatt
     4021 gtcctcgctt gggcccaaaa ggcctttttt ttttggcgcc gacgtcaggt ggcacttttc
     4081 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4141 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4201 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4261 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4321 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4381 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4441 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4501 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4561 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4621 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4681 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4741 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4801 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4861 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4921 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4981 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5041 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5101 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5161 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5221 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5281 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5341 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5401 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5461 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5521 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5581 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5641 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5701 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5761 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5821 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5881 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5941 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     6001 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     6061 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     6121 ctcattaggc accccaggc
//
