LOCUS       sequence-224912-        6123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Drosophila, Splicing acceptor, STOP, dsRED, RFP, SV40 terminator, 
            flanked attP. section marker, assemble with homology arms to make 
            Donor plasmid for CRISPR. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6123)
  AUTHORS   Zhang X, Koolhaas WH, Schnorrer F
  TITLE     A Versatile Two-Step CRISPR- and RMCE-Based Strategy for Efficient 
            Genome Engineering in Drosophila.
  JOURNAL   G3 (Bethesda). 2014 Oct 15. pii: g3.114.013979. doi: 
            10.1534/g3.114.013979.
  PUBMED    25324299
REFERENCE   2  (bases 1 to 6123)
  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: "G3 
            (Bethesda). 2014 Oct 15. pii: g3.114.013979. doi: 
            10.1534/g3.114.013979."
COMMENT     Sequence Label: pJET1.2-STOP-dsRed
FEATURES             Location/Qualifiers
     source          1..6123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    395..494
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     polyA_signal    626..760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(650..669)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     regulatory      1139..1148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1145..1147,1148..1150,1151..1825)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(1328..1348)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1546..1565
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1740..1763
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1946..2027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2055..2154
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     promoter        complement(join(3955..3961,3962..3979,3980..3985))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    4000..4021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4141..4158)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4312..4900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4392..4411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5071..5862,5863..5931))
                     /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     5694..5713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5932..6036)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6104..6122
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
       61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
      121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
      181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
      241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
      301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
      361 ttcagcaaga tacgtctcac cagtctcgaa gcttcccagg tcagaagcgg ttttcgggag
      421 tagtgcccca actggggtaa cctttgagtt ctctcagttg ggggcgtagg gtcgccgaca
      481 tgacacaagg ggttggatcc ccgggagtcg atccaacatg gcgacttgtc ccatccccgg
      541 catgtttaaa tatactaatt attcttgaac taattttaat caaccgattt atctctcttc
      601 cgcaggtcta actaactaac accgcgatcc agacatgata agatacattg atgagtttgg
      661 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      721 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      781 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      841 caaatgtggt atggctgatt atgatcaact agtctggctc gcccggggat ctaattcaat
      901 tagagactaa ttcaattaga gctaattcaa ttaggatcca agcttatcga tttcgaaccc
      961 tcgaccgccg gagtataaat agaggcgctt cgtctacgga gcgacaattc aattcaaaca
     1021 agcaaagtga acacgtcgct aagcgaaagc taagcaaata aacaagcgca gctgaacaag
     1081 ctaaacaatc ggggtaccgc tagagtcgac ggtaccgcgg gcccgggatc caccggtcgc
     1141 caccatggtg cgctcctcca agaacgtcat caaggagttc atgcgcttca aggtgcgcat
     1201 ggagggcacc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     1261 cgagggccac aacaccgtga agctgaaggt gaccaagggc ggccccctgc ccttcgcctg
     1321 ggacatcctg tccccccagt tccagtacgg ctccaaggtg tacgtgaagc accccgccga
     1381 catccccgac tacaagaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     1441 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggctgctt
     1501 catctacaag gtgaagttca tcggcgtgaa cttcccctcc gacggccccg taatgcagaa
     1561 gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa
     1621 gggcgagatc cacaaggccc tgaagctgaa ggacggcggc cactacctgg tggagttcaa
     1681 gtccatctac atggccaaga agcccgtgca gctgcccggc tactactacg tggactccaa
     1741 gctggacatc acctcccaca acgaggacta caccatcgtg gagcagtacg agcgcaccga
     1801 gggccgccac cacctgttcc tgtagcggcg gcgactctag atcataatca gccataccac
     1861 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     1921 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     1981 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgccct aggcatatgc
     2041 ggtgtgaaat attcaacccc ttgtgtcatg tcggcgaccc tacgccccca actgagagaa
     2101 ctcaaaggtt accccagttg gggcactact cccgaaaacc gcttctgacc tgggccgcgg
     2161 aacgagagac tgtttgagac gtatctttct agaagatctc ctacaatatt ctcagctgcc
     2221 atggaaaatc gatgttcttc ttttattctc tcaagatttt caggctgtat attaaaactt
     2281 atattaagaa ctatgctaac cacctcatca ggaaccgttg taggtggcgt gggttttctt
     2341 ggcaatcgac tctcatgaaa actacgagct aaatactgat gaatccccta atgattttgg
     2401 taaaaatcat taagttaagg tggatacaca tcttgtcata tgatcaaatg gtttcgcgaa
     2461 aaatcaataa tcagacaaca agatgtgcga actcgatatt ttacacgact ctctttacca
     2521 attctgcccc gaattacact taaaacgact caacagctta acgttggctt gccacgcatt
     2581 acttgactgt aaaactctca ctcttaccga acttggccgt aacctgccaa ccaaagcgag
     2641 aacaaaacat aacatcaaac gaatcgaccg attgttaggt aatcgtcacc tccacaaaga
     2701 gcgactcgct gtataccgtt ggcatgctag ctttatctgt tcgggcaata cgatgcccat
     2761 tgtacttgtt gactggtctg atattcgtga gcaaaaacga cttatggtat tgcgagcttc
     2821 agtcgcacta cacggtcgtt ctgttactct ttatgagaaa gcgttcccgc tttcagagca
     2881 atgttcaaag aaagctcatg accaatttct agccgacctt gcgagcattc taccgagtaa
     2941 caccacaccg ctcattgtca gtgatgctgg ctttaaagtg ccatggtata aatccgttga
     3001 gaagctgggt tggtactggt taagtcgagt aagaggaaaa gtacaatatg cagacctagg
     3061 agcggaaaac tggaaaccta tcagcaactt acatgatatg tcatctagtc actcaaagac
     3121 tttaggctat aagaggctga ctaaaagcaa tccaatctca tgccaaattc tattgtataa
     3181 atctcgctct aaaggccgaa aaaatcagcg ctcgacacgg actcattgtc accacccgtc
     3241 acctaaaatc tactcagcgt cggcaaagga gccatgggtt ctagcaacta acttacctgt
     3301 tgaaattcga acacccaaac aacttgttaa tatctattcg aagcgaatgc agattgaaga
     3361 aaccttccga gacttgaaaa gtcctgccta cggactaggc ctacgccata gccgaacgag
     3421 cagctcagag cgttttgata tcatgctgct aatcgccctg atgcttcaac taacatgttg
     3481 gcttgcgggc gttcatgctc agaaacaagg ttgggacaag cacttccagg ctaacacagt
     3541 cagaaatcga aacgtactct caacagttcg cttaggcatg gaagttttgc ggcattctgg
     3601 ctacacaata acaagggaag acttactcgt ggctgcaacc ctactagctc aaaatttatt
     3661 cacacatggt tacgctttgg ggaaattatg aggggatctc tcagagctaa atattcaata
     3721 tgttcctctt gaccaacttt attctgcatt ttttttgaac gaggtttaga gcaagcttca
     3781 ggaaactgag acaggaattt tattaaaaat ttaaattttg aagaaagttc agggttaata
     3841 gcatccattt tttgctttgc aagttcctca gcattcttaa caaaagacgt ctcttttgac
     3901 atgtttaaag tttaaacctc ctgtgtgaaa ttattatccg ctcataattc cacacattat
     3961 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4021 aattgcgttg cgctcactgc caattgcttt ccagtcggga aacctgtcgt gccagctgca
     4081 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4141 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4201 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4261 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4321 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4381 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4441 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4501 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4561 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4621 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4681 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4741 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4801 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4861 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4921 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4981 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5041 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5101 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5161 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5221 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5281 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5341 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5401 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5461 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5521 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5581 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5641 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5701 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5761 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5821 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5881 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5941 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6001 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6061 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6121 gcc
//
