LOCUS       sequence-324133-        4367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x8-Compact promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4367)
  AUTHORS   Donahue PS, Draut JW, Muldoon JJ, Edelstein HI, Bagheri N, Leonard 
            JN
  TITLE     The COMET toolkit for composing customizable genetic programs in 
            mammalian cells.
  JOURNAL   Nat Commun. 2020 Feb 7;11(1):779. doi: 10.1038/s41467-019-14147-5.
  PUBMED    32034124
REFERENCE   2  (bases 1 to 4367)
  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"; journalName: "Nat 
            Commun."; date: "2020-02-7"; volume: "11(1):779. doi"; pages: " 
            10.1038/s41467-019-14147-5"
COMMENT     Sequence Label: pPD292 ZF43x8-Compact_EYFP
FEATURES             Location/Qualifiers
     source          1..4367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      242..251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(248..250,251..253,254..967)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(293..314)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(554..573)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     901..922
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1012..1029)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1018..1242
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1288..1716
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1375..1394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1585..1606
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1739..1935
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1786..1921
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1848..1867
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2055..2176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2092..2111)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2146..2165
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2225..2241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2225..2241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2238..2260)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2249..2265
                     /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(2273..2279,2280..2297,2298..2303))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2318..2339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2456..2473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2627..3215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2707..2726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3386..4177,4178..4246))
                     /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     4009..4028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4247..4351)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgaatttga gaccacagtg atacggtctc
       61 cgatcggagt gaggaaaatt agagtgagga cgatcggagt gaggataatt agagtgagga
      121 cgatcggagt gaggaaaatt agagtgagga cgatcggagt gaggataatt agagtgagga
      181 cgatccctct agagggtata taatgggggc cactagtcta ctaccagagc tcatcgctag
      241 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      301 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      361 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      421 gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca
      481 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      541 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
      601 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
      661 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
      721 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
      781 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
      841 caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca
      901 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
      961 caagtaggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     1021 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1081 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1141 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1201 aagtcaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     1261 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     1321 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     1381 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1441 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1501 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1561 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1621 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1681 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     1741 aggctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     1801 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     1861 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     1921 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggagg ttgggcttcg
     1981 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     2041 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2101 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2161 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     2221 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2281 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2341 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2401 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2461 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2521 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2581 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2641 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2701 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2761 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2821 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2881 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2941 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3001 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3061 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3121 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3181 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3241 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3301 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3361 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3421 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3481 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagat ccacgctcac
     3541 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3601 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3661 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3721 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3781 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3841 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3901 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3961 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4021 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4081 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4141 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4201 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4261 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4321 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaa
//
