LOCUS       sequence-324139-        4354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x9-Compact promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4354)
  TITLE     Leonard Lab COMET plasmids
REFERENCE   2  (bases 1 to 4354)
  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"
COMMENT     Sequence Label: pPD293 ZF43x9-Compact_EYFP
FEATURES             Location/Qualifiers
     source          1..4354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      229..238
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(235..237,238..240,241..954)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(280..301)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(541..560)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     888..909
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(999..1016)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1005..1229
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1275..1703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1362..1381)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1572..1593
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1726..1922
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1773..1908
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1835..1854
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2042..2163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2079..2098)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2133..2152
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2212..2228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2212..2228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2225..2247)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2236..2252
                     /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(2260..2266,2267..2284,2285..2290))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2305..2326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2443..2460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2614..3202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2694..2713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3373..4164,4165..4233))
                     /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     3996..4015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4234..4338)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgaattaga gtgaggacga tcggagtgag
       61 gaaaattaga gtgaggacga tcggagtgag gataattaga gtgaggacga tcggagtgag
      121 gaaaattaga gtgaggacga tcggagtgag gataattaga gtgaggacga tccctctaga
      181 gggtatataa tgggggccac tagtctacta ccagagctca tcgctagcgc caccatggtg
      241 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      301 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      361 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      421 accaccttcg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
      481 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      541 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      601 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
      661 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
      721 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
      781 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
      841 agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
      901 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaggcggcc
      961 gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     1021 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     1081 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     1141 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag tcaatagcag
     1201 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     1261 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     1321 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     1381 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     1441 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     1501 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     1561 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     1621 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     1681 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg ctcaattagt
     1741 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     1801 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     1861 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     1921 aaaagctccc gggagcttgt atatccattt tcggaggttg ggcttcggaa tcgttttccg
     1981 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     2041 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2101 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2161 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     2221 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2281 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2341 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2401 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2461 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2521 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2581 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2641 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2701 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2761 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2821 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2881 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2941 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3001 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3061 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3121 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3181 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3241 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3301 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3361 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3421 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3481 accatctggc cccagtgctg caatgatacc gcgagatcca cgctcaccgg ctccagattt
     3541 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3601 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3661 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3721 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3781 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3841 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3901 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3961 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4021 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4081 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4141 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4201 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4261 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4321 acaaataggg gttccgcgca catttccccg aaaa
//
