LOCUS       sequence-322948-        4234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF43x1-Spaced promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4234)
  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 4234)
  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: pPD059 ZF43x1-Spaced_EYFP
FEATURES             Location/Qualifiers
     source          1..4234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      125..134
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(131..133,134..136,137..850)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(176..197)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(437..456)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     784..805
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(895..912)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    901..1125
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1171..1599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1258..1277)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1468..1489
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1622..1818
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1669..1804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1731..1750
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1938..2059
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1975..1994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2029..2048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2108..2124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2108..2124)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2121..2143)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2132..2148
                     /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(2156..2162,2163..2180,2181..2186))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2201..2222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2339..2356)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2510..3098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2590..2609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3269..4060,4061..4129))
                     /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     3892..3911
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4130..4234)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctga attagagtga
       61 ggacgatccc tctagagggt atataatggg ggccactagt ctactaccag agctcatcgc
      121 tagcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      181 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      241 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      301 ctggcccacc ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga
      361 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      421 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      481 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
      541 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
      601 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
      661 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
      721 cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga
      781 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
      841 gtacaagtag gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga
      901 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
      961 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     1021 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     1081 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     1141 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     1201 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1261 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1321 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1381 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1441 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1501 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1561 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     1621 gttaggctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     1681 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     1741 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     1801 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg aggttgggct
     1861 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     1921 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1981 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2041 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     2101 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2161 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2221 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2281 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2341 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2401 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2461 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2521 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2581 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2641 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2701 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2761 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2821 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2881 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2941 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3001 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3061 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3121 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3181 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3241 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3301 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3361 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gatccacgct
     3421 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3481 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3541 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3601 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3661 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3721 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3781 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3841 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3901 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3961 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4021 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4081 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4141 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4201 gtatttagaa aaataaacaa ataggggttc cgcg
//
