LOCUS       sequence-324617-        4339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the Logic 37 AND 158 NOT 43 4 repeats 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4339)
  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 4339)
  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: pPD129 AND Gate, 4 repeats
FEATURES             Location/Qualifiers
     source          1..4339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      214..223
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(220..222,223..225,226..939)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(265..286)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(526..545)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     873..894
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(984..1001)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    990..1214
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1260..1688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1347..1366)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1557..1578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1711..1907
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1758..1893
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1820..1839
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2027..2148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2064..2083)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2118..2137
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2197..2213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2197..2213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2210..2232)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2221..2237
                     /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(2245..2251,2252..2269,2270..2275))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2290..2311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2428..2445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2599..3187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2679..2698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3358..4149,4150..4218))
                     /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     3981..4000
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4219..4323)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgaatttgt agatggacag agtgaggacg
       61 tgcagagtgt agatggacag agtgaggacg tgcagagtgt agatggacag agtgaggacg
      121 tgcagagtgt agatggacag agtgaggacg tgcgatccct ctagagggta tataatgggg
      181 gccactagtc tactaccaga gctcatcgct agcgccacca tggtgagcaa gggcgaggag
      241 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      301 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
      361 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
      421 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      481 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      541 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      601 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      661 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
      721 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
      781 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
      841 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
      901 gccgggatca ctctcggcat ggacgagctg tacaagtagg cggccgctcg agtctagagg
      961 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     1021 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     1081 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     1141 ggtggggcag gacagcaagg gggaggattg ggaagtcaat agcaggcatg ctggggatgc
     1201 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     1261 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1321 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1381 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1441 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1501 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1561 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1621 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1681 cgcgaattaa ttctgtggaa tgtgtgtcag ttaggctcaa ttagtcagca accatagtcc
     1741 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     1801 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     1861 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     1921 cttgtatatc cattttcgga ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     1981 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     2041 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2101 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     2161 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2221 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2281 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2341 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2401 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2461 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2521 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2581 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2641 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2701 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2761 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2821 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2881 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2941 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3001 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3061 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3121 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3181 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3241 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3301 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3361 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3421 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3481 tgctgcaatg ataccgcgag atccacgctc accggctcca gatttatcag caataaacca
     3541 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3601 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3661 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3721 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3781 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3841 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3901 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3961 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4021 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4081 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4141 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4201 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4261 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4321 gcgcacattt ccccgaaaa
//
