LOCUS       sequence-324228-        4444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x15-Compact promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4444)
  TITLE     Leonard Lab COMET plasmids
REFERENCE   2  (bases 1 to 4444)
  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: pPD300 ZF43x15-Compact_EYFP
FEATURES             Location/Qualifiers
     source          1..4444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      319..328
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(325..327,328..330,331..1044)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(370..391)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(631..650)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     978..999
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1089..1106)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1095..1319
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1365..1793
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1452..1471)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1662..1683
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1816..2012
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1863..1998
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1925..1944
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2132..2253
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2169..2188)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2223..2242
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2302..2318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2302..2318)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2315..2337)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2326..2342
                     /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(2350..2356,2357..2374,2375..2380))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2395..2416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2533..2550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2704..3292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2784..2803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3463..4254,4255..4323))
                     /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     4086..4105
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4324..4428)
                     /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 tcggagtgag gaaaattaga gtgaggacga tcggagtgag
      241 gataattaga gtgaggacga tccctctaga gggtatataa tgggggccac tagtctacta
      301 ccagagctca tcgctagcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
      361 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      421 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      481 aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct gcagtgcttc
      541 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      601 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      661 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      721 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
      781 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
      841 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
      901 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc
      961 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1021 ggcatggacg agctgtacaa gtaggcggcc gctcgagtct agagggcccg tttaaacccg
     1081 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1141 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1201 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1261 caagggggag gattgggaag tcaatagcag gcatgctggg gatgcggtgg gctctatggc
     1321 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     1381 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1441 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1501 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1561 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1621 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1681 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     1741 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     1801 tggaatgtgt gtcagttagg ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     1861 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     1921 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     1981 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     2041 tcggaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     2101 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     2161 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2221 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     2281 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2341 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2401 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2461 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3121 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3601 gcgagatcca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3721 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3781 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3841 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3901 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3961 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4021 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4081 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4141 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4201 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4261 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4321 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4381 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4441 aaaa
//
