LOCUS       sequence-318352-        4234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EYFP reporter vector for the ZF92x1 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: pPD090 ZF92x1_EYFP
FEATURES             Location/Qualifiers
     source          1..4234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     215..234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(453..557)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      682..691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(688..690,691..693,694..1407)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(733..754)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(994..1013)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1341..1362
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1452..1469)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1458..1682
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1728..2156
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1815..1834)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2025..2046
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2179..2375
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2226..2361
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2288..2307
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2495..2616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2532..2551)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2586..2605
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2665..2681)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2665..2681)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2678..2700)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2689..2705
                     /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(2713..2719,2720..2737,2738..2743))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2758..2779
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2896..2913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3067..3655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3147..3166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3826..4234,1..383,384..452))
                     /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"
ORIGIN
        1 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
       61 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
      121 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
      181 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
      241 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
      301 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
      361 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
      421 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
      481 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
      541 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
      601 gatctgaatt agatgtagcc tgatccctct agagggtata taatgggggc cactagtcta
      661 ctaccagagc tcatcgctag cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
      721 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      781 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      841 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgcagtgc
      901 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      961 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1021 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1081 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1141 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     1201 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1261 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac
     1321 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1381 ctcggcatgg acgagctgta caagtaggcg gccgctcgag tctagagggc ccgtttaaac
     1441 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1501 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1561 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1621 cagcaagggg gaggattggg aagtcaatag caggcatgct ggggatgcgg tgggctctat
     1681 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1741 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1801 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1861 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1921 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1981 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2041 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2101 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2161 ctgtggaatg tgtgtcagtt aggctcaatt agtcagcaac catagtcccg cccctaactc
     2221 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2281 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     2341 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     2401 ttttcggagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     2461 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     2521 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2581 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     2641 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2701 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2761 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2821 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2881 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2941 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3001 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3061 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3121 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3181 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3241 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3301 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3361 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3421 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3481 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3541 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3601 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3661 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3721 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3781 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3841 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3901 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3961 accgcgagat ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4021 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4081 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4141 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4201 acgatcaagg cgagttacat gatcccccat gttg
//
