LOCUS       sequence-306359-        3845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pBSII-KS(-)-based vector backbone containing a dominant eye marker 
            for constructing Drosophila CRISPR HDR templates. Marker allows 
            counter selection against vector integration events..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3845)
  AUTHORS   Nyberg KG, Nguyen JQ, Kwon YJ, Blythe S, Beitel GJ, Carthew RW
  TITLE     A Pipeline for Precise and Efficient Genome Editing by sgRNA-Cas9 
            RNPs in Drosophila.
  JOURNAL   Fly (Austin). 2020 Oct 5. doi: 10.1080/19336934.2020.1832416.
  PUBMED    33016195
REFERENCE   2  (bases 1 to 3845)
  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: "Fly 
            (Austin). 2020 Oct 5. doi: 10.1080/19336934.2020.1832416."
COMMENT     Sequence Label: pBS-GMR-eya(shRNA)
FEATURES             Location/Qualifiers
     source          1..3845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward 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"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     677..693
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(727..743)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(728..744)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        983..1221
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     1168..1189
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     primer_bind     complement(1657..1677)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1657..1675)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1696..1712)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1696..1712)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1709..1731)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1720..1736
                     /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(1744..1750,1751..1768,1769..1774))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1789..1810
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1927..1944)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2098..2686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2178..2197)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2857..3648,3649..3717))
                     /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     3480..3499
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3718..3822)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa
      721 gcttatcgat accgtcgacc tcgagggggg gcccggtact cgcgagcgat cgcctcgagc
      781 ccagtggaaa cccttgaaat gcctttaagt cgagcccagt ggaaaccctt gaaatgcctt
      841 taagtcgagc ccagtggaaa cccttgaaat gcctttaagt cgagcccagt ggaaaccctt
      901 gaaatgcctt taagtcgagc ccagtggaaa cccttgaaat gcctttaagt cgactctagg
      961 ttaattaatc gcgaagcgag cgccggagta taaatagagg cgcttcgtct acggagcgac
     1021 aattcaattc aaacaagcaa agtgaacacg tcgctaagcg aaagctaagc aaataaacaa
     1081 gcgcagctga acaagctaaa caatctgcag taaagtgcaa gttaaagtga atcaattaaa
     1141 agtaaccagc aaccaagtaa atcaactgca actactgaaa tctgccaaga agtaattatt
     1201 gaatacaaga agagaactct gaatagggaa ttgggaattg agatctgttc tagaaaacat
     1261 cccataaaac atcccatatt cagccgctag cagtaacatg actactacta ctacatagtt
     1321 atattcaagc atatgtagta gtagtagtca tgttgcgaat tcaggcgaga catcggagtt
     1381 gaaactaaaa ctgaattact agagtggaca tatgcaccta ggactagagc aaactagttc
     1441 tgatctgcta gacaattgtt ggcatcaggt aggcatcaca cacgattaac aacccctaaa
     1501 aatacacttt gaaaatattg aaaatatgtt tttgtataca tttttgatat tttcaaataa
     1561 tacgcagtta taaaactcat tagctaaccc attttttctt tgcttatgct tacagattgc
     1621 aaagaactag agccgcgggt accccagctt ttgttccctt tagtgagggt taattgcgcg
     1681 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1741 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1801 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1861 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1921 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1981 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2041 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2101 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2161 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2221 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2281 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2341 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2401 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2461 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2521 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2581 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2641 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2701 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2761 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2821 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2881 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2941 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3001 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3061 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3121 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3181 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3241 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3301 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3361 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3421 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3481 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3541 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3601 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3661 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3721 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3781 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3841 gccac
//
