LOCUS       sequence-99766-e        5313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4xnrUAS driving GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5313)
  AUTHORS   Akitake CM, Macurak M, Halpern ME, Goll MG
  TITLE     Transgenerational analysis of transcriptional silencing in 
            zebrafish.
  JOURNAL   Dev Biol. 2011 Apr 15;352(2):191-201. doi: 
            10.1016/j.ydbio.2011.01.002. Epub 2011 Jan 9.
  PUBMED    21223961
REFERENCE   2  (bases 1 to 5313)
  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"; doi: 
            "10.1016/j.ydbio.2011.01.002"; journalName: "Dev Biol"; date: 
            "2011-04-15- 15"; volume: "352"; issue: "2"; pages: "191-201"
COMMENT     Sequence Label: 4xnrUAS:GFP
FEATURES             Location/Qualifiers
     source          1..5313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(659..670,671..833)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     1181..1197
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1181..1197
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    1314..1448
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1351..1370)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1405..1424
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(1493..2206,2207..2209,2210..2212))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1496..2206,2207..2209,2210..2212))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1538..1559)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1887..1906
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2146..2167
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2209..2218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2496..2512)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2497..2513)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    join(2908..3095,3096..3107)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3125..3145)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3125..3143)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3164..3180)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3164..3180)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3177..3199)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3188..3204
                     /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(3212..3218,3219..3236,3237..3242))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3257..3278
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3395..3412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3566..4154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3646..3665)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4325..5116,5117..5185))
                     /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     4948..4967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5186..5290)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 gaggtgtaaa aagtactcaa aaattttact caagtgaaag tacaagtact tagggaaaat
      721 tttactcaat taaaagtaaa agtatctggc tagaatctta cttgagtaaa agtaaaaaag
      781 tactccatta aaattgtact tgagtattaa ggaagtaaaa gtaaaagcaa gaaagaaaac
      841 tagagattct tgtttaagct tttaatctca aaaaacatta aatgaaatgc atacaaggtt
      901 ttatcctgct ttagaactgt ttgtatttaa ttatcaaact ataagacaga caatctaatg
      961 ccagtacacg ctactcaaag ttgtaaaacc tcagatttaa cttcagtaga agctgattct
     1021 caaaattgtt agtgtcaagc ctagctcttt tggggctgaa aagcaatcct gcagtgctga
     1081 aaagcctctc acaggcagcc gatgcgggaa gaggtgtatt agtcttgata gagaggctgc
     1141 aaatagcagg aaacgtgagc agagacagat ctgcggccgc tctagaacta gtggatctga
     1201 tctagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1261 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1321 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1381 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     1441 tctggatcat catcgatgaa ttaattcgag ctccaccgcg gtggcggccg ctttacttgt
     1501 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     1561 gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg tagtggttgt
     1621 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     1681 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
     1741 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     1801 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     1861 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     1921 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     1981 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc
     2041 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     2101 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     2161 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggcga
     2221 ccggtggatc cacgcgtctt cgaggtcgag ggaattcgtg tggaggagct caaagtgagg
     2281 ctgagacgcg atgggatcca ttatataccc tctagagtcg acccttgcac gcgtcggagg
     2341 acagtactcc gaggaactct acggaagact ctcctccgcc tgtgaagacg gagggctgtc
     2401 acccgctagg actcacggat tagaagccac cgtggaggcc taagcttgat ggccgcgtga
     2461 tcccccgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgacct cgaggggggg
     2521 cccgcgggag acagaggaca catttcatct tgaatgaatc tttatttaca ccacttaaag
     2581 aaaagaattc gcgaagatac ggccacgggt gctcttgatc ctgtggctga ttttggactg
     2641 tgctgctcgc agctgctgat gaatcacata cttcctccat tttcttccac tgattgactg
     2701 ttataatttc cctaatttcc aggtcaaggt gctgtgcatt gtggtaatag atgtgacatg
     2761 acgtcacttc caaaggacca atgaacatgt ctgaccaatt tcatataatg tgaaaacgat
     2821 tttcataggc agaataaata acatttaaat taaactgggc atcagcgcaa ttcaattggt
     2881 ttggtaatag caagggaaaa tagaatgaag tgatctccaa aaaataagta ctttttgact
     2941 gtaaataaaa ttgtaaggag taaaaagtac ttttttttct aaaaaaatgt aattaagtaa
     3001 aagtaaaagt attgattttt aattgtactc aagtaaagta aaaatcccca aaaataatac
     3061 ttaagtacag taatcaagta aaattactca agtactttac acctctgggt accagctttt
     3121 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     3181 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3241 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3301 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3361 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3421 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3481 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3541 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3601 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3661 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3721 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3781 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3841 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3901 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3961 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4021 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4081 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4141 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4201 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4261 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4321 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4381 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4441 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4501 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4561 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4621 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4681 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4741 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4801 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4861 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4921 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4981 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5041 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5101 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5161 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5221 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5281 gggttccgcg cacatttccc cgaaaagtgc cac
//
