LOCUS       sequence-324294-        4312 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF158x3-Compact promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4312)
  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 4312)
  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: pPD922 ZF158x3-Compact_EYFP
FEATURES             Location/Qualifiers
     source          1..4312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      187..196
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(193..195,196..198,199..912)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(238..259)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(499..518)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     846..867
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(957..974)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    963..1187
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1233..1661
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1320..1339)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1530..1551
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1684..1880
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1731..1866
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1793..1812
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2000..2121
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2037..2056)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2091..2110
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2170..2186)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2170..2186)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2183..2205)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2194..2210
                     /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(2218..2224,2225..2242,2243..2248))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2263..2284
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2401..2418)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2572..3160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2652..2671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3331..4122,4123..4191))
                     /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     3954..3973
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4192..4296)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgcgagtca gtgcagacgg caagtggtcg
       61 agtgccactt gccgtctgca cgatctgtag atggagagcc tgtagatgga ggccgtgtag
      121 atggagacgc tctctagagg gtatataatg ggggccacta gtctactacc agagctcatc
      181 gctagcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      241 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      301 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      361 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc
      421 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      481 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      541 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      601 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
      661 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
      721 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      781 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
      841 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
      901 ctgtacaagt aggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc
      961 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     1021 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     1081 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     1141 ttgggaagtc aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     1201 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     1261 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     1321 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     1381 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     1441 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     1501 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     1561 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     1621 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     1681 cagttaggct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     1741 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     1801 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     1861 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggaggttggg
     1921 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     1981 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     2041 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2101 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     2161 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     2221 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     2281 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2341 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2401 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2461 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2521 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2581 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2641 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2701 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2761 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2821 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2881 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2941 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3001 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3061 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3121 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3181 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3241 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3301 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3361 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3421 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg
     3481 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3541 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3601 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3661 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3721 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3781 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3841 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3901 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3961 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4021 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4081 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4141 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4201 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4261 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aa
//
