LOCUS       sequence-324519-        4315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF158x6-Compact promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4315)
  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 4315)
  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: pPD183 ZF158x6-Compact_EYFP v1
FEATURES             Location/Qualifiers
     source          1..4315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      190..199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(196..198,199..201,202..915)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(241..262)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(502..521)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     849..870
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(960..977)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    966..1190
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1236..1664
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1323..1342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1533..1554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1687..1883
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1734..1869
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1796..1815
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2003..2124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2040..2059)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2094..2113
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2173..2189)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2173..2189)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2186..2208)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2197..2213
                     /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(2221..2227,2228..2245,2246..2251))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2266..2287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2404..2421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2575..3163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2655..2674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3334..4125,4126..4194))
                     /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     3957..3976
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4195..4299)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgaattacg atctgtagat ggagaatttg
       61 tagatggagg atcggtagat ggaaaattag tagatggacg atcggtagat ggataattag
      121 tagatggacg atccctctag agggtatata atgggggcca ctagtctact accagagctc
      181 atcgctagcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      241 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      301 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      361 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac
      421 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      481 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      541 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
      601 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
      661 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
      721 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
      781 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
      841 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
      901 gagctgtaca agtaggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
      961 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     1021 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     1081 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     1141 ggattgggaa gtcaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     1201 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     1261 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     1321 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     1381 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     1441 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     1501 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     1561 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     1621 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     1681 tgtcagttag gctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     1741 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     1801 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     1861 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggaggtt
     1921 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     1981 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     2041 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2101 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     2161 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2221 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2281 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2341 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2401 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2461 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2521 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2581 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2641 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2701 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2761 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2821 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2881 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2941 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3001 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3061 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3121 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3181 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3241 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3301 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3361 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3421 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagatcc
     3481 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3541 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3601 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3661 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3721 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3781 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3841 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3901 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3961 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4021 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4081 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4141 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4201 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4261 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaa
//
