LOCUS       sequence-323307-        3658 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  puromycin resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3658)
  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 3658)
  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: pPAC
FEATURES             Location/Qualifiers
     source          1..3658
                     /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(816..834)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(817..834)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(852..868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(852..868)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(865..887)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    876..892
                     /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(900..906,907..924,925..930))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    945..966
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1083..1100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1254..1842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1334..1353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2013..2804,2805..2873))
                     /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     2636..2655
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2874..2978)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3056..3511)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3193..3214)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3405..3424
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(3637..3658,1)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3651..3658,1..10)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(3652..3658,1..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 cgaagcttag
      121 gctcctggct tacgggtcat acaccaggtg cgtgggccct cgggcacctc cacatcggcc
      181 gtcacggtga agcccagacg ctcgtagaag ggcagattgc gtggggcact ggtttccagg
      241 aaggctggca ctccagcacg ctcggcagct tccactcctg gcagcaccac ggcactgccc
      301 aggcccttgc cctggtggtc ggggctcacg cccacggtgg ccaggaacca gacgggctcc
      361 tttggacggt gtggggccag caggccctcc atctgctgct gggcagccag acgggagcca
      421 ctcagctcgg ccatacgggg tccaatctcg gcgaacacgg ctccagcctc cacgctctcg
      481 ggggtggtcc acacggccac ggcagctcca tcatcggcca cccagacctt gccgatatcc
      541 aggcccacgc gtgtcaggaa cagctcctgg agctcggtca cacgctcaat gtgacggtct
      601 gggtccacgg tgtgacgggt ggctgggtaa tcggcgaagg cagcagccag ggttcgcacg
      661 gcacggggca catcatcacg ggtggccaga cgcaccgtgg gcttatactc ggtcattcga
      721 gaccactcat cgcccaatca ctagtgaatt cgcggccgcc tgcaggtcga ccatatggga
      781 gagctcccaa cgcgttggat gcatagcttg agtattctat agtgtcacct aaatagcttg
      841 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
      901 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
      961 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1021 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1081 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1141 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1201 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1261 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1321 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1381 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1441 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1501 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1561 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1621 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1681 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1741 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1801 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1861 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1921 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1981 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2041 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2101 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgcgaccca
     2161 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2221 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2281 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2341 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     2401 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     2461 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     2521 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2581 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2641 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2701 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2761 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2821 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2881 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2941 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3001 cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     3061 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     3121 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     3181 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     3241 aaagggcgaa aaaccgtcta tcagggcgct ggcccactac gtgaaccatc accctaatca
     3301 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3361 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3421 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3481 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     3541 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     3601 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaac
//
