LOCUS       sequence-324222-        4435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x14-Compact promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4435)
  TITLE     Leonard Lab COMET plasmids
REFERENCE   2  (bases 1 to 4435)
  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: pPD299 ZF43x14-Compact_EYFP
FEATURES             Location/Qualifiers
     source          1..4435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      310..319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(316..318,319..321,322..1035)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(361..382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(622..641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     969..990
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1080..1097)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1086..1310
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1356..1784
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1443..1462)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1653..1674
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1807..2003
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1854..1989
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1916..1935
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2123..2244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2160..2179)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2214..2233
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2293..2309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2293..2309)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2306..2328)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2317..2333
                     /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(2341..2347,2348..2365,2366..2371))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2386..2407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2524..2541)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2695..3283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2775..2794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3454..4245,4246..4314))
                     /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     4077..4096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4315..4419)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgaattacg atcggagtga ggataattag
       61 agtgaggacg atcggagtga ggaaaattag agtgaggacg atcggagtga ggataattag
      121 agtgaggacg atcggagtga ggaaaattag agtgaggacg atcggagtga ggataattag
      181 agtgaggacg atcggagtga ggaaaattag agtgaggacg atcggagtga ggataattag
      241 agtgaggacg atccctctag agggtatata atgggggcca ctagtctact accagagctc
      301 atcgctagcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      361 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      421 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      481 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac
      541 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      601 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      661 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
      721 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
      781 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
      841 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
      901 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
      961 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1021 gagctgtaca agtaggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
     1081 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     1141 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     1201 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     1261 ggattgggaa gtcaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     1321 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     1381 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     1441 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     1501 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     1561 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     1621 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     1681 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     1741 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     1801 tgtcagttag gctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     1861 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     1921 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     1981 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggaggtt
     2041 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     2101 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     2161 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2221 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     2281 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2341 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2401 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2461 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2521 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2581 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2641 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2701 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2761 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2821 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2881 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2941 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3001 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3061 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3121 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3181 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3241 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3301 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3481 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3541 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagatcc
     3601 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3661 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3721 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3781 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3841 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3901 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3961 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4021 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4081 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4141 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4201 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4261 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4321 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4381 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaa
//
