LOCUS       sequence-324504-        4348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF158x7-Compact promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4348)
  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 4348)
  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: pPD926 ZF158x7-Compact_EYFP
FEATURES             Location/Qualifiers
     source          1..4348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      223..232
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(229..231,232..234,235..948)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(274..295)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(535..554)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     882..903
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(993..1010)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    999..1223
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1269..1697
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1356..1375)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1566..1587
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1720..1916
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1767..1902
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1829..1848
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2036..2157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2073..2092)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2127..2146
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2206..2222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2206..2222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2219..2241)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2230..2246
                     /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(2254..2260,2261..2278,2279..2284))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2299..2320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2437..2454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2608..3196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2688..2707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3367..4158,4159..4227))
                     /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     3990..4009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4228..4332)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgcgagtca gtgcagacgg caagtggtcg
       61 atgtagatgg agcgactgta gatggagacg ctgtagatgg aggatctgta gatggaggct
      121 ttgtagatgg agagcctgta gatggaggcc gtgtagatgg agacgctctc tagagggtat
      181 ataatggggg ccactagtct actaccagag ctcatcgcta gcgccaccat ggtgagcaag
      241 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      301 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      361 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      421 ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
      481 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      541 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      601 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      661 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
      721 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
      781 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac
      841 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
      901 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaggc ggccgctcga
      961 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     1021 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1081 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1141 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagtcaata gcaggcatgc
     1201 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     1261 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1321 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1381 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1441 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     1501 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1561 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     1621 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     1681 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt taggctcaat tagtcagcaa
     1741 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     1801 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     1861 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     1921 tcccgggagc ttgtatatcc attttcggag gttgggcttc ggaatcgttt tccgggacgc
     1981 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     2041 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2101 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     2161 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     2221 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2281 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2341 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     2401 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2461 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2521 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2581 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2641 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2701 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2761 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2821 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2881 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2941 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3001 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3061 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3121 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3181 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3241 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3301 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3361 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3421 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3481 tggccccagt gctgcaatga taccgcgaga tccacgctca ccggctccag atttatcagc
     3541 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3601 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3661 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3721 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3781 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3841 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3901 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3961 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4021 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4081 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4141 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4201 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4261 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4321 aggggttccg cgcacatttc cccgaaaa
//
