LOCUS       sequence-254621-        4273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPaint universal donor plasmid for gene exon insertion. Encodes 
            3xP3-GFP visible eye marker and multiple cloning site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4273)
  AUTHORS   Bosch JA, Colbeth R, Zirin J, Perrimon N
  TITLE     Gene Knock-Ins in Drosophila Using Homology-Independent Insertion of
            Universal Donor Plasmids.
  JOURNAL   Genetics. 2019 Nov 4. pii: genetics.119.302819. doi: 
            10.1534/genetics.119.302819.
  PUBMED    31685521
REFERENCE   2  (bases 1 to 4273)
  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: "Genetics. 
            2019 Nov 4. pii: genetics.119.302819. doi: 
            10.1534/genetics.119.302819."
COMMENT     Sequence Label: pCRISPaint-3xP3-GFP
FEATURES             Location/Qualifiers
     source          1..4273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     653..673
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        655..673
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(692..725)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    755..876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(766..785)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     820..839
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      1713..1722
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1948..1981)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2112..2131)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2113..2131)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2141..2158)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2141..2157)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2150..2172)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2298..2754)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2436..2457)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2648..2667
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2781..2885
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2886..2954,2955..3746)
                     /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(3104..3123)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3917..4273,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
      661 ccctcactaa agggaacaaa agctggagct cataacttcg tatagcatac attatacgaa
      721 gttatcgtat gggccttcgc tgcttacaga cggttaagat acattgatga gtttggacaa
      781 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
      841 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
      901 atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa
      961 tgtggtatgg ctgattatga tctagagtcg cggcccctac ttgtacagct catccatgcc
     1021 cagggtaatg ccggcggcgg tcacaaactc cagcagcacc atgtggtcgc gcttctcgtt
     1081 gggatccttg ctcagggcgc tctgggtgct caggtaatgg ttatcgggca gcagcactgg
     1141 gccgtcgccg attggggtgt tctgctggta gtgatcggcc agctgcacgc cgccatcctc
     1201 gatgttgtgg cggatcttga agttggcctt gatgccgttc ttctgcttgt cggccatgat
     1261 gtacacgttg tggctgttgt agttgtactc cagcttgtgg cccaggatgt ttccatcctc
     1321 cttgaaatcg atgcccttca gctcgatgcg gttcacgagg gtatcgccct cgaacttcac
     1381 ttcggcgcgg gtcttgtagt tgccgtcatc cttgaaaaag atggtgcgct cctgcacgta
     1441 gccctcgggc atggcgctct tgaagaaatc gtgctgcttc atgtgatcgg ggtagcggct
     1501 gaagcactgc acgccatagg tcagggtggt cacgagggtt ggccatggca cgggcagctt
     1561 gccggtggtg cagatgaact tcagggtcag ctttccatag gtggcgtcgc cctcgccctc
     1621 gccggacacg ctgaacttgt ggccgttcac gtcgccatcc agctccacca gaattggcac
     1681 cacgccggtg aacagctcct cgcccttgga caccatggtg gcgaccggct tcgagccgat
     1741 tgtttagctt gttcagctgc gcttgtttat ttgcttagct ttcgcttagc gacgtgttca
     1801 ctttgcttgt ttgaattgaa ttgtcgctcc gtagacgaag cgcctctatt tatactccgg
     1861 cggtcgaggg ttcgaaatcg ataagcttgg atcctaattg aattagctct aattgaatta
     1921 gtctctaatt gaattagatc ccgtacgata acttcgtata gcatacatta tacgaagtta
     1981 tagatctgtg aattctaggt accgtctcga gcagctagcg tgcggccgcg ctacctccgc
     2041 ttccaccgga ccccccgctt tttgggtact ggcccggcgc gcctcaggtc gtgaagcttg
     2101 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     2161 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     2221 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     2281 gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2341 ttgttaaatc agctcatttt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2401 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2461 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2521 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2581 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2641 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2701 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     2761 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2821 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2881 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2941 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3001 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3061 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3121 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3181 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3241 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3301 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3361 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3421 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3481 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3541 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3601 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3661 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3721 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3781 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3841 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3901 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3961 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4021 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4081 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4141 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4201 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4261 gcttggagcg aac
//
