LOCUS       sequence-324569-        4309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x6-Compact(overlapping ZF37) 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4309)
  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 4309)
  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: pPD270 ZF43x6-Compact(overlapping ZF37)_EYFP
FEATURES             Location/Qualifiers
     source          1..4309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      171..180
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(177..179,180..182,183..896)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(222..243)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(483..502)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     830..851
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(941..958)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    947..1171
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1217..1645
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1304..1323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1514..1535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1668..1864
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1715..1850
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1777..1796
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1984..2105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2021..2040)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2075..2094
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2154..2170)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2154..2170)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2167..2189)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2178..2194
                     /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(2202..2208,2209..2226,2227..2232))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2247..2268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2385..2402)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2556..3144)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2636..2655)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3315..4106,4107..4175))
                     /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     3938..3957
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4176..4280)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctg aattggagtg aggacgtgta gagtgaggac gtgcggagtg
       61 aggacgtgta gagtgaggac gtgcggagtg aggacgtgta gagtgaggac gtgccctcta
      121 gagggtatat aatgggggcc actagtctac taccagagct catcgctagc gccaccatgg
      181 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      241 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
      301 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
      361 tgaccacctt cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc
      421 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
      481 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
      541 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
      601 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
      661 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
      721 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
      781 tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
      841 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaggcgg
      901 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
      961 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     1021 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     1081 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agtcaatagc
     1141 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     1201 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1261 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     1321 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1381 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     1441 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1501 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     1561 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     1621 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta ggctcaatta
     1681 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     1741 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     1801 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     1861 caaaaagctc ccgggagctt gtatatccat tttcggaggt tgggcttcgg aatcgttttc
     1921 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     1981 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2041 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2101 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     2161 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2221 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2281 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     2341 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2401 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2461 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2521 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2581 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2641 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2701 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2761 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2821 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2881 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2941 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3001 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3061 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3121 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3181 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3241 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3301 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3361 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3421 ttaccatctg gccccagtgc tgcaatgata ccgcgagatc cacgctcacc ggctccagat
     3481 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3541 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3601 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3661 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3721 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3781 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3841 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3901 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3961 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4021 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4081 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4141 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4201 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4261 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacg
//
