LOCUS       sequence-323508-        4433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x7-Spaced promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4433)
  TITLE     Leonard Lab COMET plasmids
REFERENCE   2  (bases 1 to 4433)
  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: pPD088 ZF43x7-Spaced_EYFP
FEATURES             Location/Qualifiers
     source          1..4433
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      324..333
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(330..332,333..335,336..1049)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(375..396)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(636..655)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     983..1004
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1094..1111)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1100..1324
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1370..1798
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1457..1476)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1667..1688
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1821..2017
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1868..2003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1930..1949
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2137..2258
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2174..2193)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2228..2247
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2307..2323)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2307..2323)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2320..2342)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2331..2347
                     /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(2355..2361,2362..2379,2380..2385))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2400..2421
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2538..2555)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2709..3297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2789..2808)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3468..4259,4260..4328))
                     /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     4091..4110
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4329..4433)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctga attcgagtga
       61 ggacaatcgt atacaagtag atccagagtg aggactacct atagaagtaa gagtgaggac
      121 aatcgtatac aagtagatcc ctctagaggg agatccagag tgaggactac ctatagaagt
      181 aagagtgagg acaatcgtat acaagtagat ccagagtgag gactacctat agaagtaaga
      241 gtgaggacaa tcgtatacaa gtagatccct ctagagggta tataatgggg gccactagtc
      301 tactaccaga gctcatcgct agcgccacca tggtgagcaa gggcgaggag ctgttcaccg
      361 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      421 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      481 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgcagt
      541 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      601 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
      661 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
      721 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
      781 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
      841 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
      901 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag
      961 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1021 ctctcggcat ggacgagctg tacaagtagg cggccgctcg agtctagagg gcccgtttaa
     1081 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     1141 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     1201 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     1261 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     1321 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     1381 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1441 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     1501 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     1561 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     1621 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     1681 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     1741 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     1801 ttctgtggaa tgtgtgtcag ttaggctcaa ttagtcagca accatagtcc cgcccctaac
     1861 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     1921 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     1981 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     2041 cattttcgga ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     2101 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     2161 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2221 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     2281 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2341 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2401 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2461 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2521 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2581 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2641 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2701 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2761 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2821 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2881 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2941 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3001 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3061 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3121 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3181 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3241 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3301 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3361 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3421 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3481 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3541 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3601 ataccgcgag atccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3661 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3721 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3781 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3841 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3901 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3961 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4021 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4081 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4141 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4201 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4261 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4321 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4381 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcg
//
