LOCUS       sequence-136548-        4440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  template base vector for N-terminal tagging, constitutive Puromycin 
            resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4440)
  AUTHORS   Kunzelmann S, Bottcher R, Schmidts I, Forstemann K
  TITLE     A Comprehensive Toolbox for Genome Editing in Cultured Drosophila 
            melanogaster Cells.
  JOURNAL   G3 (Bethesda). 2016 Jun 1;6(6):1777-85. doi: 10.1534/g3.116.028241.
  PUBMED    27172193
REFERENCE   2  (bases 1 to 4440)
  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"; doi: "10.1534/g3.116"; 
            journalName: "G3 (Bethesda)"; date: "2016-06-1- 1"; volume: "6"; 
            issue: "6"; pages: "1777-85"
COMMENT     Sequence Label: pRB35
FEATURES             Location/Qualifiers
     source          1..4440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(372..404)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(1738..1771)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        complement(join(2272..2278,2279..2296,2297..2302))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2317..2338
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2458..2475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2629..3217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2709..2728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3388..4179,4180..4248))
                     /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     4011..4030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4249..4353)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4421..4439
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
       61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
      121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
      181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
      241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
      301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
      361 ttcagcaaga taagttccta tactttctag agaataggaa cttccatatg ggcgcgccct
      421 gttggaatat actattcaac ctacaaaaat aacgttaaac aacactactt tatatttgat
      481 atgaatggcc acacctttta tgccataaaa catattgtaa gagaatacca ctctttttat
      541 tccttctttc cttcttgtac gttttttgct gtgagtaggt cgtggtgctg gtgttgcagt
      601 tgaaataact taaaatataa atcataaaac tcaaacataa acttgactat ttatttattt
      661 attaagaaag gaggtaccgg tggcgcgcca tgaccgagta caagcctaca gtgcgactgg
      721 ccacacgcga tgacgttccc agagccgttc ggacgttggc tgccgcgttt gcggactatc
      781 ccgcaactcg ccatactgtg gaccctgatc gccacatcga gagggtgact gagcttcagg
      841 agctgttcct gaccagggta gggctggaca taggcaaggt ctgggtggcc gacgatggag
      901 ccgctgttgc ggtgtggact accccagaga gcgtggaagc aggggctgtc tttgccgaaa
      961 ttgggccaag gatggccgaa ctgtctgggt cccgtttggc tgcccaacag cagatggaag
     1021 gactgcttgc cccacatcga cccaaggagc ctgcatggtt cctcgctacc gtgggtgtga
     1081 gcccagatca ccagggcaaa ggtctgggat cagccgtcgt tctgcctggc gtcgaggctg
     1141 ctgagagagc aggagtaccg gcattcctcg aaaccagtgc tccacggaac ctcccgttct
     1201 acgagagact gggctttacg gtcacagccg atgtggaagt gcccgaaggc cctagaacct
     1261 ggtgcatgac acggaaaccc ggtgcatgaa aattaaatat aaataaagct attaaaataa
     1321 attactatct aacatatgcc taggcttgtt gatttgtttt ttgtttattt actgcggtaa
     1381 ttcctagtct tagaagctta ttgaacttgt tgatgccttc tggttgggaa atcctcctct
     1441 ttatacccga tccagtccaa tcgaaactgt gggccgtggt gcccgataag ataagaaagc
     1501 cgtgtgcaag tcgatcggcg tgcgcaaaag cataatgctt tttctgcccg gcgatgataa
     1561 gcgatttgca cacgccctga taacaggaat tgattttttt ggtgcgcaaa gcagtgaaag
     1621 ctctataaga atgctggacc gccagtcgac aatcatcaaa cagtgctagc taacggctaa
     1681 gtgaatcaag tttattttaa tataatcaac aacaaaatat tttaacacaa acctagggaa
     1741 gttcctatac tttctagaga ataggaactt ctaatccaaa atgggatcca ccggtgaatt
     1801 cggtacctct ccagctgctc caagcggcgg tactagtatc tttctagaag atctcctaca
     1861 atattctcag ctgccatgga aaatcgatgt tcttctttta ttctctcaag attttcaggc
     1921 tgtatattaa aacttatatt aagaactatg ctaaccacct catcaggaac cgttgtaggt
     1981 ggcgtgggtt ttcttggcaa tcgactctca tgaaaactac gagctaaata ttcaatatgt
     2041 tcctcttgac caactttatt ctgcattttt tttgaacgag gtttagagca agcttcagga
     2101 aactgagaca ggaattttat taaaaattta aattttgaag aaagttcagg gttaatagca
     2161 tccatttttt gctttgcaag ttcctcagca ttcttaacaa aagacgtctc ttttgacatg
     2221 tttaaagttt aaacctcctg tgtgaaatta ttatccgctc ataattccac acattatacg
     2281 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2341 tgcgttgcgc tcactgccaa ttgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2401 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2461 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3061 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4381 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
//
