LOCUS       sequence-317946-        4234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF97x1 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: pPD092 ZF97x1_EYFP
FEATURES             Location/Qualifiers
     source          1..4234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(96..524)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(206..227)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     418..437
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    570..794
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     783..800
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(845..1558,1559..1561,1562..1564))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(890..911)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1239..1258
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1498..1519
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1561..1570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        1695..1799
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1800..1868,1869..2660)
                     /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     complement(2018..2037)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2831..3419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3320..3339
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3573..3590
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3707..3728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3743..3748,3749..3766,3767..3773)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3781..3797
                     /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)."
     primer_bind     3786..3808
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3805..3821
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3805..3821
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3870..3991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3881..3900)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3935..3954
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(join(4111..4234,1..73))
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      join(4125..4234,1..26)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4179..4198)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
       61 taattgagcc taactgacac acattccaca gaattaattc gcgttaaatt tttgttaaat
      121 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
      181 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
      241 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
      301 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
      361 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
      421 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
      481 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat accccctaga
      541 gccccagctg gttctttccg cctcagaagc catagagccc accgcatccc cagcatgcct
      601 gctattgact tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga
      661 atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg
      721 agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca
      781 actagaaggc acagtcgagg ctgatcagcg ggtttaaacg ggccctctag actcgagcgg
      841 ccgcctactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
      901 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tggtagctca
      961 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     1021 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga
     1081 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
     1141 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     1201 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
     1261 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     1321 gctgcttcat gtggtcgggg tagcgggcga agcactgcag gccgtagccg aaggtggtca
     1381 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
     1441 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt
     1501 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     1561 ccatggtggc gctagcgatg agctctggta gtagactagt ggcccccatt atataccctc
     1621 tagagggatc tctcccataa taattcagat ctcccgatcc gtcgacgtca ggtggcactt
     1681 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1741 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1801 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1861 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1921 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1981 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2041 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2101 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2161 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2221 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2281 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2341 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2401 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2461 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggatctc
     2521 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2581 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2641 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     2701 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2761 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2821 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2881 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2941 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3001 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3061 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3121 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3181 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3241 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3301 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3361 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3421 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3481 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3541 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3601 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     3661 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     3721 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     3781 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
     3841 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
     3901 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3961 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
     4021 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaacctcc
     4081 gaaaatggat atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc
     4141 tcctcactac ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa
     4201 aaaaattagt cagccatggg gcggagaatg ggcg
//
