LOCUS       sequence-313250-        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      11..466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(98..117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     308..329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     592..614
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     606..623
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     607..623
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     633..652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        633..651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     684..700
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     684..700
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(734..750)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(735..751)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        990..1228
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     1175..1196
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     primer_bind     complement(1664..1684)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1664..1682)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1703..1719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1703..1719)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1716..1738)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1727..1743
                     /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(1751..1757,1758..1775,1776..1781))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1796..1817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1934..1951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2105..2693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2185..2204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2864..3655,3656..3724))
                     /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     3487..3506
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3725..3829)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
       61 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
      121 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
      181 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
      241 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
      301 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
      361 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
      421 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca
      481 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
      541 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
      601 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg
      661 agctccaccg cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaattcg
      721 atatcaagct tatcgatacc gtcgacctcg agggggggcc cggtactcgc gagcgatcgc
      781 ctcgagccca gtggaaaccc ttgaaatgcc tttaagtcga gcccagtgga aacccttgaa
      841 atgcctttaa gtcgagccca gtggaaaccc ttgaaatgcc tttaagtcga gcccagtgga
      901 aacccttgaa atgcctttaa gtcgagccca gtggaaaccc ttgaaatgcc tttaagtcga
      961 ctctaggtta attaatcgcg aagcgagcgc cggagtataa atagaggcgc ttcgtctacg
     1021 gagcgacaat tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa gctaagcaaa
     1081 taaacaagcg cagctgaaca agctaaacaa tctgcagtaa agtgcaagtt aaagtgaatc
     1141 aattaaaagt aaccagcaac caagtaaatc aactgcaact actgaaatct gccaagaagt
     1201 aattattgaa tacaagaaga gaactctgaa tagggaattg ggaattgaga tctgttctag
     1261 aaaacatccc ataaaacatc ccatattcag ccgctagcag taacatgact actactacta
     1321 catagttata ttcaagcata tgtagtagta gtagtcatgt tgcgaattca ggcgagacat
     1381 cggagttgaa actaaaactg aattactaga gtggacatat gcacctagga ctagagcaaa
     1441 ctagttctga tctgctagac aattgttggc atcaggtagg catcacacac gattaacaac
     1501 ccctaaaaat acactttgaa aatattgaaa atatgttttt gtatacattt ttgatatttt
     1561 caaataatac gcagttataa aactcattag ctaacccatt ttttctttgc ttatgcttac
     1621 agattgcaaa gaactagagc cgcgggtacc ccagcttttg ttccctttag tgagggttaa
     1681 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1741 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1801 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1861 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1921 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1981 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2041 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2101 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2161 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2221 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2281 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2341 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2401 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2461 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2521 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2581 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2641 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2701 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2761 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2821 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2881 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2941 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3001 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3061 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3121 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3181 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3241 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3301 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3361 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3421 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3481 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3541 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3601 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3661 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3721 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3781 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3841 gaaaa
//
