LOCUS       sequence-401601-        3865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes PhiYFP, 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 PhiYFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3865)
  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 3865)
  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-PhiYFP-pA
FEATURES             Location/Qualifiers
     source          1..3865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(173..964,965..1033))
                     /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     796..815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1034..1138)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1165..1620
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1252..1271)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1462..1483
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1751..1799
                     /note="synthetic polyadenylation signal"
     misc_feature    1813..1904
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1853..1872
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     primer_bind     1918..1934
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1918..1934
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2004..2708
                     /codon_start=1
                     /product="Phialidium yellow fluorescent proteim"
                     /label=PhiYFP
                     /note="mammalian codon-optimized"
                     /translation="MSSGALLFHGKIPYVVEMEGDVDGHTFSIRGKGYGDASVGKVDAQ
                     FICTTGDVPVPWSTLVTTLTYGAQCFAKYGPELKDFYKSCMPDGYVQERTITFEGDGNF
                     KTRAEVTFENGSVYNRVKLNGQGFKKDGHVLGKNLEFNFTPHCLYIWGDQANHGLKSAF
                     KICHEIAGSKGDFIVADHTQMNTPIGGGPVHVPEYHHMSYHVKLSKDVTDHRDNMSLTE
                     TVRAVDCRKTYL"
     polyA_signal    2734..2855
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2745..2764)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2799..2818
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3108..3125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3279..3865,1..2))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3359..3378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
       61 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      121 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
      181 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
      241 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
      301 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
      361 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
      421 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
      481 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
      541 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
      601 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
      661 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
      721 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
      781 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
      841 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
      901 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
      961 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1021 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1081 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1141 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1201 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1261 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1321 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     1381 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     1441 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     1501 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     1561 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     1621 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     1681 tattacgcca gcccaagcta ccatgataag taagtaatat taaggtacgg gaggtacttg
     1741 gagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     1801 atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa
     1861 ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggatcccag
     1921 gaaacagcta tgacgagctc tctagcgggg ggctataaaa gggggtgggg gcgttcgtcc
     1981 tgctatctag cgtcgcgttg accatgagca gcggcgccct gctgttccac ggcaagatcc
     2041 cctacgtggt ggagatggag ggcgatgtgg atggccacac cttcagcatc cgcggtaagg
     2101 gctacggcga tgccagcgtg ggcaaggtgg atgcccagtt catctgcacc accggcgatg
     2161 tgcccgtgcc ctggagcacc ctggtgacca ccctgaccta cggcgcccag tgcttcgcca
     2221 agtacggccc cgagctgaag gatttctaca agagctgcat gcccgatggc tacgtgcagg
     2281 agcgcaccat caccttcgag ggcgatggca atttcaagac ccgcgccgag gtgaccttcg
     2341 agaatggcag cgtgtacaat cgcgtgaagc tgaatggcca gggcttcaag aaggatggcc
     2401 acgtgctggg caagaatctg gagttcaatt tcacccccca ctgcctgtac atctggggcg
     2461 atcaggccaa tcacggcctg aagagcgcct tcaagatctg ccacgagatc gccggcagca
     2521 agggcgattt catcgtggcc gatcacaccc agatgaatac ccccatcggc ggcggccccg
     2581 tgcacgtgcc cgagtaccac cacatgagct accacgtgaa gctgagcaag gatgtgaccg
     2641 atcaccgcga taatatgagc ctgacggaga ccgtgcgcgc cgtggattgc cgcaagacct
     2701 acctgtaagg ccggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     2761 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2821 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     2881 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     2941 gtggtaaaat cgataaggat cgagtatgtc gaccgatgcc cttgagagcc ttcaacccag
     3001 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     3061 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
     3121 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3181 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3241 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3301 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3361 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3421 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3481 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3541 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3601 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3661 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3721 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3781 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3841 attacgcgca gaaaaaaagg atctc
//
