LOCUS       sequence-323933-        4264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x3-Compact promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4264)
  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 4264)
  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: pPD287 ZF43x3-Compact_EYFP
FEATURES             Location/Qualifiers
     source          1..4264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      139..148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(145..147,148..150,151..864)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(190..211)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(451..470)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     798..819
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(909..926)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    915..1139
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1185..1613
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1272..1291)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1482..1503
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1636..1832
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1683..1818
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1745..1764
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1952..2073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1989..2008)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2043..2062
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2122..2138)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2122..2138)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2135..2157)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2146..2162
                     /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(2170..2176,2177..2194,2195..2200))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2215..2236
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2353..2370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2524..3112)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2604..2623)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3283..4074,4075..4143))
                     /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     3906..3925
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4144..4248)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgaattaga gtgaggacga tcggagtgag
       61 gataattaga gtgaggacga tccctctaga gggtatataa tgggggccac tagtctacta
      121 ccagagctca tcgctagcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
      181 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      241 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      301 aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct gcagtgcttc
      361 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      421 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      481 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      541 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
      601 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
      661 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
      721 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc
      781 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
      841 ggcatggacg agctgtacaa gtaggcggcc gctcgagtct agagggcccg tttaaacccg
      901 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
      961 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1021 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1081 caagggggag gattgggaag tcaatagcag gcatgctggg gatgcggtgg gctctatggc
     1141 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     1201 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1261 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1321 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1381 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1441 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1501 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     1561 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     1621 tggaatgtgt gtcagttagg ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     1681 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     1741 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     1801 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     1861 tcggaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     1921 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     1981 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2041 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     2101 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2161 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2221 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2281 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2341 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2401 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2461 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2521 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2581 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2641 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2701 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2761 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2821 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2881 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2941 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3001 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3061 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3121 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3181 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3241 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3301 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3361 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3421 gcgagatcca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3481 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3541 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3601 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3661 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3721 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3781 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3841 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3901 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3961 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4021 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4081 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4141 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4201 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4261 aaaa
//
