LOCUS       sequence-340309-        2699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of Puro resistance gene (pac) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2699)
  TITLE     Verstreken lab plasmids
REFERENCE   2  (bases 1 to 2699)
  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"
COMMENT     Sequence Label: pIVTR(T7G)-PuroR
FEATURES             Location/Qualifiers
     source          1..2699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      101..110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             107..709
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     606..626
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     rep_origin      complement(944..1532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1024..1043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1703..2494,2495..2563))
                     /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     2326..2345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2564..2668)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taatacgact cactataggg acatttgctt ctgacacaac tgtgttcact agcaacctca
       61 aacagaattc gagctcggta cccggggatc ctctagagtc gccaccatgg ccaccgagta
      121 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
      181 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
      241 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
      301 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
      361 gggggcggtg ttcgccgaga tcggctcgcg catggccgag ttgagcggtt cccggctggc
      421 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
      481 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
      541 gctccccgga gtggaggcgg ccgagcgcgc tggggtgccc gccttcctgg agacctccgc
      601 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
      661 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag ttcgcgtctg
      721 gaacagacct gcaggcatgc aagcttgctc gctttcttgc tgtccaattt ctattaaagg
      781 ttcctttgtt ccctaagtcc aactactaaa ctgggggata ttatgaaggg ccttgagcat
      841 ctggattctg cctaataaaa aacatttatt ttcattgcaa aaccatggga gcaaaaggcc
      901 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      961 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1021 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1081 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1141 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1201 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1261 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1321 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1381 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1441 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1501 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1561 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1621 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1681 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1741 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1801 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1861 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     1921 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     1981 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2041 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2101 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2161 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2221 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2281 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     2341 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     2401 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     2461 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2521 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     2581 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2641 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
