LOCUS       sequence-335079-        3433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zeocin/Phleomycin resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3433)
  AUTHORS   Matinyan N, Karkhanis MS, Gonzalez Y, Jain A, Saltzman A, 
            Malovannaya A, Sarrion-Perdigones A, Dierick HA, Venken KJT
  TITLE     Multiplexed drug-based selection and counterselection genetic 
            manipulations in Drosophila.
  JOURNAL   Cell Rep. 2021 Sep 14;36(11):109700. doi: 
            10.1016/j.celrep.2021.109700.
  PUBMED    34525356
REFERENCE   2  (bases 1 to 3433)
  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: "Cell Rep.";
            date: "2021-09-14"; volume: "36(11):109700. doi"; pages: " 
            10.1016/j.celrep.2021.109700"
COMMENT     Sequence Label: pBLE
FEATURES             Location/Qualifiers
     source          1..3433
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..36
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        17..35
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      66..75
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      70..79
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(591..609)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(592..609)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(627..643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(627..643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(640..662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    651..667
                     /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(675..681,682..699,700..705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    720..741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(858..875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1029..1617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1109..1128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1788..2579,2580..2648))
                     /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     2411..2430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2649..2753)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2831..3286)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2968..2989)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3180..3199
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(3412..3433,1)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3426..3433,1..10)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(3427..3433,1..10)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacggccagt gaattgtaat acgactcact atagggcgaa ttgggcccga cgtcgcatgc
       61 tcccggccgc catggcggcc gcgggaattc gattgggcga tgagtggtct cgaatggcca
      121 agctgaccag tgccgtgcca gtgctgacgg ctcgtgatgt ggcaggagcc gtggagttct
      181 ggaccgaccg tctgggcttc agtcgcgatt tcgtggagga tgatttcgca ggcgtggtcc
      241 gtgacgatgt gaccctgttc atctcggccg tgcaggacca ggtggtgccc gataacaccc
      301 tggcctgggt ctgggtccgt ggcctggatg agctctacgc cgagtggagc gaggtggtgt
      361 ccaccaactt ccgagatgcc agtggaccag caatgaccga gattggcgag cagccgtggg
      421 gacgcgagtt cgccctccgt gacccagcag gcaactgcgt gcacttcgtg gccgaggagc
      481 aggactaagc ttcgagacca ctcatcgccc aatcactagt gaattcgcgg ccgcctgcag
      541 gtcgaccata tgggagagct cccaacgcgt tggatgcata gcttgagtat tctatagtgt
      601 cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
      661 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
      721 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
      781 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
      841 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
      901 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
      961 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1021 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1081 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1141 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1201 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1261 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1321 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1381 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1441 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     1501 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     1561 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1621 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     1681 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     1741 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     1801 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     1861 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     1921 ataccgcgcg acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     1981 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2041 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2101 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2161 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2221 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2281 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     2341 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2401 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     2461 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     2521 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     2581 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     2641 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     2701 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     2761 ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     2821 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2881 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2941 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3001 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgctggccc actacgtgaa
     3061 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3121 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3181 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3241 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca
     3301 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     3361 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     3421 gacgttgtaa aac
//
