LOCUS       sequence-324512-        4363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF158x8-Compact promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4363)
  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 4363)
  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: pPD927 ZF158x8-Compact_EYFP
FEATURES             Location/Qualifiers
     source          1..4363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      238..247
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(244..246,247..249,250..963)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(289..310)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(550..569)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     897..918
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1008..1025)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1014..1238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1284..1712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1371..1390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1581..1602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1735..1931
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1782..1917
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1844..1863
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2051..2172
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2088..2107)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2142..2161
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2221..2237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2221..2237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2234..2256)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2245..2261
                     /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(2269..2275,2276..2293,2294..2299))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2314..2335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2452..2469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2623..3211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2703..2722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3382..4173,4174..4242))
                     /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     4005..4024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4243..4347)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgcgagtca gtgcagacgg caagtggtcg
       61 atgtagatgg aggtgatgta gatggagcga ctgtagatgg agacgctgta gatggaggat
      121 ctgtagatgg aggctttgta gatggagagc ctgtagatgg aggccgtgta gatggagacg
      181 ctctctagag ggtatataat gggggccact agtctactac cagagctcat cgctagcgcc
      241 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      301 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      361 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      421 accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
      481 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      541 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      601 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
      661 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
      721 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
      781 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
      841 cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
      901 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
      961 taggcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     1021 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1081 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1141 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaagt
     1201 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     1261 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1321 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     1381 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     1441 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     1501 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     1561 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     1621 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     1681 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggc
     1741 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     1801 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     1861 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     1921 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggaggttgg gcttcggaat
     1981 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     2041 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2101 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2161 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     2221 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2281 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2341 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2401 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2461 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2521 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2581 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2641 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2701 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2761 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2821 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2881 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2941 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3001 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3061 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3121 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3181 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3241 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3301 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3361 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3421 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3481 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagatccac gctcaccggc
     3541 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3601 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3661 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3721 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3781 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3841 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3901 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3961 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4021 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4081 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4141 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4201 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4261 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4321 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaa
//
