LOCUS       sequence-320112-        4240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for COMET promoters, expresses EYFP reporter upon promoter 
            activation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4240)
  AUTHORS   Donahue PS, Draut JW, Muldoon JJ, Edelstein HI, Bagheri N, Leonard 
            JN
  TITLE     The COMET toolkit for composing customizable genetic programs in 
            mammalian cells.
  JOURNAL   Nat Commun. 2020 Feb 7;11(1):779. doi: 10.1038/s41467-019-14147-5.
  PUBMED    32034124
REFERENCE   2  (bases 1 to 4240)
  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 
            Commun."; date: "2020-02-7"; volume: "11(1):779. doi"; pages: " 
            10.1038/s41467-019-14147-5"
COMMENT     Sequence Label: pPD027 Reporter Template v1 (Near Spaced)
FEATURES             Location/Qualifiers
     source          1..4240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      131..140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(137..139,140..142,143..856)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(182..203)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(443..462)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     790..811
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(901..918)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    907..1131
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1177..1605
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1264..1283)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1474..1495
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1628..1824
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1675..1810
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1737..1756
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1944..2065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1981..2000)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2035..2054
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2114..2130)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2114..2130)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2127..2149)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2138..2154
                     /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(2162..2168,2169..2186,2187..2192))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2207..2228
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2345..2362)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2516..3104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2596..2615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3275..4066,4067..4135))
                     /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     3898..3917
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4136..4240)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctga attcgagacc
       61 acaggtctcg gatccctcta gagggtatat aatgggggcc actagtctac taccagagct
      121 catcgctagc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      181 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      241 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      301 cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgcagtgct tcgcccgcta
      361 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      421 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      481 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      541 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
      601 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
      661 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      721 gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa
      781 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      841 cgagctgtac aagtaggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag
      901 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
      961 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     1021 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     1081 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     1141 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     1201 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     1261 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     1321 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     1381 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     1441 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     1501 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     1561 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     1621 gtgtcagtta ggctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     1681 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1741 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     1801 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggaggt
     1861 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     1921 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     1981 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2041 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     2101 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2161 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2221 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2281 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2341 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2401 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2461 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2521 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2581 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2641 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2701 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2761 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2821 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2881 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2941 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3001 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3061 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3121 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3181 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3241 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3301 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3361 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagatc
     3421 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3481 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3541 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3601 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3661 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3721 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3781 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3841 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3901 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3961 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4021 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4081 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4141 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4201 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
//
