LOCUS       sequence-401600-        3880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes mCerulean, which can be expressed with the activation of a 
            synthetic promoter by Cas enzymes. A cloning site is provided to 
            construct the synthetic Cas-targeted promoter upstream of 
            mCerulean..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3880)
  AUTHORS   Kang H, Fitch JC, Varghese RP, Thorne CA, Cusanovich DA
  TITLE     Optimization of a Cas12a-Driven Synthetic Gene Regulatory Network 
            System.
  JOURNAL   ACS Synth Biol. 2025 May 16;14(5):1732-1744. doi: 
            10.1021/acssynbio.5c00084. Epub 2025 May 2.
  PUBMED    40316310
REFERENCE   2  (bases 1 to 3880)
  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: 
            "10.1021/acssynbio.5c00084"; journalName: "ACS Synth Biol"; date: 
            "2025-05-16- 16"; volume: "14"; issue: "5"; pages: "1732-1744"
COMMENT     Sequence Label: Px-mCerulean-pA
FEATURES             Location/Qualifiers
     source          1..3880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(169..960,961..1029))
                     /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     792..811
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1030..1134)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1161..1616
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1248..1267)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1458..1479
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1747..1795
                     /note="synthetic polyadenylation signal"
     misc_feature    1809..1900
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1849..1868
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     primer_bind     1914..1930
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1914..1930
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(2000..2002,2003..2005,2006..2719)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2045..2066)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2306..2325)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2653..2674
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2745..2866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2756..2775)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2810..2829
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3119..3136)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3290..3878)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3370..3389)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
       61 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      121 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      181 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      241 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      301 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      361 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      421 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      481 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      541 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      601 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
      661 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
      721 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
      781 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
      841 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
      901 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
      961 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1021 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1081 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1141 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1201 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1261 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1321 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1381 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1441 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1501 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1561 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttgcca
     1621 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     1681 acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg tacttggagc
     1741 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     1801 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
     1861 gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgatagga tcccaggaaa
     1921 cagctatgac gagctctcta gcggggggct ataaaagggg gtgggggcgt tcgtcctgct
     1981 atctagcgtc gcgttgacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2041 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2101 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2161 gcccgtgccc tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt gcttcgcccg
     2221 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2281 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2341 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2401 cggcaacatc ctggggcaca agctggagta caacgccatc agcgacaacg tctatatcac
     2461 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     2521 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2581 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga
     2641 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2701 ggacgagctg tacaagtaag gccggccgct tcgagcagac atgataagat acattgatga
     2761 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2821 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2881 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     2941 cctctacaaa tgtggtaaaa tcgataagga tcgagtatgt cgaccgatgc ccttgagagc
     3001 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     3061 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct
     3121 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3181 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3241 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3301 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3361 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3421 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3481 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3541 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3601 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3661 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3721 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3781 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3841 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
//
