LOCUS       sequence-486432-        4592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative control, luciferase cassette deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4592)
  AUTHORS   Concepcion D, Liang C, Kim D, Hamilton BA
  TITLE     Nonequivalence of Zfp423 premature termination codons in mice.
  JOURNAL   bioRxiv [Preprint]. 2025 Jun 2:2025.05.30.656936. doi: 
            10.1101/2025.05.30.656936.
  PUBMED    40501617
REFERENCE   2  (bases 1 to 4592)
  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: "bioRxiv 
            [Preprint]. 2025 Jun 2:2025.05.30.656936. doi: 
            10.1101/2025.05.30.656936."
COMMENT     Sequence Label: pZNpF-deleted
FEATURES             Location/Qualifiers
     source          1..4592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1046..1065
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1213..1812
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1213..1232)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1709..1729
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1839..1887
                     /note="synthetic polyadenylation signal"
     CDS             join(2153..2221,2222..3013)
                     /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     complement(2371..2390)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3026..3162
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3207..3278
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3504..4092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3584..3603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(4592,1..1209)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc
       61 cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg
      121 gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga
      181 cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa
      241 aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga
      301 ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg
      361 cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg
      421 ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca
      481 agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca
      541 gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg
      601 tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg
      661 ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac
      721 gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat
      781 ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg
      841 acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt aggtgtgcgg
      901 taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga
      961 caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt
     1021 ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg
     1081 gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata
     1141 tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt
     1201 ttgttagaca acatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     1261 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     1321 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     1381 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     1441 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     1501 gagttgagcg gttcccggct ggccgcgcag caacagatgg agggcctcct ggcgccgcac
     1561 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     1621 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg ccgccgagcg cgccggggtg
     1681 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     1741 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     1801 cccggtgcct aggctagtat gtaagcctag tcttagataa taaaatcgct atccatcgaa
     1861 gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac
     1921 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa
     1981 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca
     2041 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt
     2101 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgagtat
     2161 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2221 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg
     2281 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     2341 ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     2401 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2461 ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2521 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc
     2581 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2641 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2701 aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag cttcccggca
     2761 acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2821 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2881 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2941 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3001 taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct
     3061 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     3121 ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc
     3181 aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag
     3241 gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct
     3301 atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc
     3361 gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct
     3421 ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3481 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3541 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3601 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3661 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3721 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3781 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3841 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3901 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3961 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4021 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4081 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4141 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4201 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4261 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4321 atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt
     4381 cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc
     4441 ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat
     4501 ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag
     4561 atccgaggaa tatcgaaatc ggggcgcgcc tg
//
