LOCUS       sequence-324332-        4333 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 4333)
  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 4333)
  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: pPD925 ZF158x6-Compact_EYFP
FEATURES             Location/Qualifiers
     source          1..4333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      208..217
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(214..216,217..219,220..933)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(259..280)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(520..539)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     867..888
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(978..995)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    984..1208
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1254..1682
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1341..1360)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1551..1572
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1705..1901
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1752..1887
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1814..1833
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2021..2142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2058..2077)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2112..2131
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2191..2207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2191..2207)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2204..2226)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2215..2231
                     /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(2239..2245,2246..2263,2264..2269))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2284..2305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2422..2439)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2593..3181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2673..2692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3352..4143,4144..4212))
                     /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     3975..3994
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4213..4317)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctgcgagtca gtgcagacgg caagtggtcg
       61 atgtagatgg agacgctgta gatggaggat ctgtagatgg aggctttgta gatggagagc
      121 ctgtagatgg aggccgtgta gatggagacg ctctctagag ggtatataat gggggccact
      181 agtctactac cagagctcat cgctagcgcc accatggtga gcaagggcga ggagctgttc
      241 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      301 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
      361 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg
      421 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      481 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
      541 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
      601 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
      661 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
      721 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
      781 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
      841 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
      901 atcactctcg gcatggacga gctgtacaag taggcggccg ctcgagtcta gagggcccgt
      961 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     1021 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     1081 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     1141 gcaggacagc aagggggagg attgggaagt caatagcagg catgctgggg atgcggtggg
     1201 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     1261 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1321 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1381 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1441 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1501 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1561 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1621 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1681 ttaattctgt ggaatgtgtg tcagttaggc tcaattagtc agcaaccata gtcccgcccc
     1741 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     1801 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     1861 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     1921 tatccatttt cggaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     1981 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     2041 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2101 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     2161 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2221 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2281 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2341 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2401 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2461 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2521 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2581 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2641 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2701 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2761 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2821 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2881 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2941 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3001 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3061 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3121 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3181 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3241 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3301 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3361 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3421 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3481 aatgataccg cgagatccac gctcaccggc tccagattta tcagcaataa accagccagc
     3541 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3601 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3661 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3721 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3781 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3841 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3901 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3961 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4021 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4081 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4141 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4201 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4261 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4321 atttccccga aaa
//
