LOCUS       sequence-324624-        4309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the Logic 37 AND 43 AND 158 2 repeats 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4309)
  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 4309)
  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: pPD187 Tri-AND Gate, 2 repeats
FEATURES             Location/Qualifiers
     source          1..4309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      184..193
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(190..192,193..195,196..909)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(235..256)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(496..515)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     843..864
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(954..971)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    960..1184
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1230..1658
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1317..1336)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1527..1548
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1681..1877
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1728..1863
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1790..1809
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1997..2118
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2034..2053)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2088..2107
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2167..2183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2167..2183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2180..2202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2191..2207
                     /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(2215..2221,2222..2239,2240..2245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2260..2281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2398..2415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2569..3157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2649..2668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3328..4119,4120..4188))
                     /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     3951..3970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4189..4293)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgaattagt agatggaacc tcagaggacg
       61 tgacctcaga gtgaggaacc tcagtagatg gaacctcaga ggacgtgacc tcagagtgag
      121 gaagatccct ctagagggta tataatgggg gccactagtc tactaccaga gctcatcgct
      181 agcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      241 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      301 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      361 tggcccaccc tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac
      421 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
      481 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
      541 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
      601 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
      661 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
      721 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
      781 gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
      841 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
      901 tacaagtagg cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
      961 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1021 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     1081 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     1141 ggaagtcaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     1201 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     1261 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1321 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1381 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1441 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1501 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1561 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     1621 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     1681 ttaggctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     1741 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     1801 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     1861 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga ggttgggctt
     1921 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     1981 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2041 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2101 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     2161 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2221 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2281 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2341 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2401 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2461 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2521 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2581 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2641 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2701 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2761 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2821 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2881 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2941 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3001 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3061 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3121 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3181 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3241 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3301 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3361 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3421 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag atccacgctc
     3481 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3541 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3601 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3661 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3721 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3781 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3841 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3901 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3961 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4021 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4081 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4141 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4201 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4261 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaa
//
