LOCUS       sequence-96175-e        4465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green beetle luciferase with nuclear localization signal (NLS) and 
            ClpXP degron tag (ssrA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4465)
  AUTHORS   Mazo-Vargas A, Park H, Aydin M, Buchler NE
  TITLE     Measuring fast gene dynamics in single cells with time-lapse 
            luminescence microscopy
  JOURNAL   Mol Biol Cell. 2014 Sep 17. pii: mbc.E14-07-1187.
  PUBMED    25232010
REFERENCE   2  (bases 1 to 4465)
  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: "Mol Biol 
            Cell. 2014 Sep 17. pii: mbc.E14-07-1187."
COMMENT     Sequence Label: pNB774
FEATURES             Location/Qualifiers
     source          1..4465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(111..902,903..971))
                     /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     734..753
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(972..1063)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1105..1124
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1130..1149
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1368..1388
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3578..3595)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3578..3594)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3693..3710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3864..4452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3944..3963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcttttctac ggggtctgac gctcagtgga acgacgcgcg cgtaactcac gttaagggat
       61 tttggtcatg agcttgcgcc gtcccgtcaa gtcagcgtaa tgctctgctt ttaccaatgc
      121 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      181 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagcgctgcg
      241 atgataccgc gagaaccacg ctcaccggct ccggatttat cagcaataaa ccagccagcc
      301 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      361 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      421 atcgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      481 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      541 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      601 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      661 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      721 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      781 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      841 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      901 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
      961 tgaatactca tattcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1021 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt cagtgttaca
     1081 accaattaac caattctgaa cattatcgcg agcccattta tacctgaata tggctcataa
     1141 caccccttgt ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     1201 agaagtgaaa cgccgtagcg ccgatggtag tgtggggact ccccatgcga gagtagggaa
     1261 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgcccgggct
     1321 aattatgggg tgtcgccctt cgctgaattt aattaaccat ggctccacca aaaaaaaaaa
     1381 gaaaagttgc catgggtggg ggcagcatga tgaagagaga aaaaaatgtt gtttacggcc
     1441 ccgaacccaa acaccctttg ggaaacttta ctgctggaga aatgctctac aatgcccttc
     1501 ataagcattc ccacataccg caagcaatat tagatgtgat gggtaatgaa tcgctgtcat
     1561 atcaagaatt tttcgacact actgtcaagc taggacaaag tcttcaaaat tgtggataca
     1621 agatgaatga tgtagtgtcg atctgtgctg agaacaataa aagatttttc atccccatta
     1681 tttcagcttg gtatattggt atggttgtag cacctgttaa tgaagactac atcccagatg
     1741 aactttgtaa agtcacgggt atatcaaaac caatactggt cttcactaca agaaaaatct
     1801 tacctaaggt attagaggta aaggacagaa ctaattacat aaagagaatt ataatactag
     1861 attctgaaga aaatctgctt ggttgcgaaa gtcttcataa ttttatgtcc cgttattcgg
     1921 ataataatct tcagaccttt aagcctctac attacgatcc tgttgatcaa gtggcagcta
     1981 ttttatgttc gtcaggcaca actggattac ccaaaggtgt tatgcaaacg catagaaaca
     2041 tttgtgttcg acttacacat gcttcggatc ccagagttgg aacacaactt attcctggag
     2101 tatcagtttt ggcgtatctg cctttcttcc atgcttttgg gttttctata aacttaggat
     2161 actttatggt gggccttcgt gttgttatgc taagacgatt taatcaagaa gtatttttaa
     2221 aagccattca agattatgaa gttcgaagtg taatcaacgt tccatcaaca atactgttct
     2281 tgtcgaaaag tcctttagtt gacaaatacg atttatcgac tttggcggaa ttgtgttgcg
     2341 gcgctgcacc attagcaaag gaagttgctg agatagcagt gaaacgacta aacctgccag
     2401 gaattcgttg tggatatggt ttgacagagt ctacttcagc taatatacat actcttcaca
     2461 atgaatttaa gtcaggatca cttggaaaag tcactcctta tatggctgcg aaaataatag
     2521 ataggaacac tggtgaagct ttgggaccaa atcaagttgg agaactatgc atctggggtc
     2581 ctatggtaac aaaaggttac gtgaacaatc cacaagccac caaagaggct attgatgacg
     2641 acggttggct tcactctgga gactttggat actatgatga ggatgaatat ttctatatag
     2701 tggaccgtta caaggaactt attaaatata aaggctatca ggtagcacct gtagaattag
     2761 aagagatttt attacaacat ccaggtataa gagatgttgc tgtcgttggt attcctgata
     2821 tagaagctgg agaactacca gctgggttcg tggttaaaca acccggagca caacttacag
     2881 caaaagaagt ttacgatttt cttgcccaac gggtctctca ttcaaagtat ttgcgtggag
     2941 gagttcgatt cgttgattca atacccagga atgttacagg taaaatttca agaaaagaac
     3001 ttcgagaggc gttgatggaa aaagctggag gtggcagtat gtacaaaact agtgcagcga
     3061 acgacgaaaa ttacgccctt gcagcgtaat aactagttaa tacaaataag gcgcgcccgt
     3121 caaaagggcg acacaaaatt tattctaaat gcataataaa tactgataac atcttatagt
     3181 ttgtattata ttttgtatta tcgttgacat gtataatttt gatatcaaaa actgattttc
     3241 cctttattat tttcgagatt tattttctta attctcttta acaaactaga aatattgtat
     3301 atacaaaaaa tcataaataa tagatgaata gtttaattat aggtgttcat caatcgaaaa
     3361 agcaacgtat cttatttaaa gtgcgttgct tttttctcat ttataaggtt aaataattct
     3421 catatatcaa gcaaagtgac aggcgccctt aaatattctg acaaatgctc tttccctaaa
     3481 ctccccccat aaaaaaaccc gccgaagcgg gtttttacgt tatttgcgga ttaacgatta
     3541 ctcgttatca gaaccgccca gggggcccga gcttaagact ggccgtcgtt ttacaacaca
     3601 gaaagagttt gtagaaacgc aaaaaggcca tccgtcaggg gccttctgct tagtttgatg
     3661 cctggcagtt ccctactctc gccttccgct tcctcgctca ctgactcgct gcgctcggtc
     3721 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3781 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3841 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3901 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3961 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4021 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4081 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4141 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4201 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4261 acagagttct tgaagtggtg ggctaactac ggctacacta gaagaacagt atttggtatc
     4321 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4381 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4441 aaaggatctc aagaagatcc tttga
//
