LOCUS       sequence-324141-        4384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x11-Compact promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4384)
  TITLE     Leonard Lab COMET plasmids
REFERENCE   2  (bases 1 to 4384)
  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: pPD295 ZF43x11-Compact_EYFP
FEATURES             Location/Qualifiers
     source          1..4384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      259..268
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(265..267,268..270,271..984)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(310..331)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(571..590)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     918..939
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1029..1046)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1035..1259
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1305..1733
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1392..1411)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1602..1623
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1756..1952
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1803..1938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1865..1884
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2072..2193
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2109..2128)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2163..2182
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2242..2258)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2242..2258)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2255..2277)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2266..2282
                     /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(2290..2296,2297..2314,2315..2320))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2335..2356
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2473..2490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2644..3232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2724..2743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3403..4194,4195..4263))
                     /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     4026..4045
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4264..4368)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgaattaga gtgaggacga tcggagtgag
       61 gataattaga gtgaggacga tcggagtgag gaaaattaga gtgaggacga tcggagtgag
      121 gataattaga gtgaggacga tcggagtgag gaaaattaga gtgaggacga tcggagtgag
      181 gataattaga gtgaggacga tccctctaga gggtatataa tgggggccac tagtctacta
      241 ccagagctca tcgctagcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
      301 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      361 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      421 aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct gcagtgcttc
      481 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      541 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      601 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      661 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
      721 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
      781 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
      841 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc
      901 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
      961 ggcatggacg agctgtacaa gtaggcggcc gctcgagtct agagggcccg tttaaacccg
     1021 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1081 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1141 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1201 caagggggag gattgggaag tcaatagcag gcatgctggg gatgcggtgg gctctatggc
     1261 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     1321 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1381 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1441 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1501 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1561 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1621 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     1681 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     1741 tggaatgtgt gtcagttagg ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     1801 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     1861 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     1921 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     1981 tcggaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     2041 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     2101 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2161 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     2221 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2281 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2341 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2401 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2461 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2521 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2581 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2641 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2701 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2761 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2821 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2881 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2941 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3001 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3061 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3121 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3181 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3241 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3301 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3361 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3421 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3481 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3541 gcgagatcca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3601 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3661 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3721 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3781 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3841 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3901 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3961 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4021 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4081 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4141 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4201 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4261 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4321 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4381 aaaa
//
