LOCUS       sequence-324515-        4399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF158x12-Compact promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4399)
  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 4399)
  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: pPD928 ZF158x12-Compact_EYFP
FEATURES             Location/Qualifiers
     source          1..4399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      274..283
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(280..282,283..285,286..999)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(325..346)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(586..605)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     933..954
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1044..1061)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1050..1274
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1320..1748
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1407..1426)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1617..1638
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1771..1967
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1818..1953
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1880..1899
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2087..2208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2124..2143)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2178..2197
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2257..2273)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2257..2273)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2270..2292)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2281..2297
                     /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(2305..2311,2312..2329,2330..2335))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2350..2371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2488..2505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2659..3247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2739..2758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3418..4209,4210..4278))
                     /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     4041..4060
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4279..4383)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgcgagtgt agatggagct ggtgtagatg
       61 gagtcgatgt agatggagga cttgtagatg gagtcgatgt agatggaggt gatgtagatg
      121 gagcgactgt agatggagac gctgtagatg gaggatctgt agatggaggc tttgtagatg
      181 gagagcctgt agatggaggc cgtgtagatg gagacgctct ctagagggta tataatgggg
      241 gccactagtc tactaccaga gctcatcgct agcgccacca tggtgagcaa gggcgaggag
      301 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      361 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
      421 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
      481 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      541 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      601 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      661 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      721 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
      781 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
      841 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
      901 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
      961 gccgggatca ctctcggcat ggacgagctg tacaagtagg cggccgctcg agtctagagg
     1021 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     1081 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     1141 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     1201 ggtggggcag gacagcaagg gggaggattg ggaagtcaat agcaggcatg ctggggatgc
     1261 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     1321 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1381 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1441 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1501 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1561 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1621 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1681 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1741 cgcgaattaa ttctgtggaa tgtgtgtcag ttaggctcaa ttagtcagca accatagtcc
     1801 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     1861 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     1921 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     1981 cttgtatatc cattttcgga ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     2041 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     2101 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2161 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     2221 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2281 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2341 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2401 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2461 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2521 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2581 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2641 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2701 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2761 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2821 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2881 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2941 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3001 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3061 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3121 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3181 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3241 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3301 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3361 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3421 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3481 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3541 tgctgcaatg ataccgcgag atccacgctc accggctcca gatttatcag caataaacca
     3601 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3661 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3721 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3781 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3841 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3901 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3961 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4021 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4081 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4141 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4201 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4261 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4321 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4381 gcgcacattt ccccgaaaa
//
