LOCUS       sequence-273970-        2981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low Phosphate inducible Nef.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2981)
  AUTHORS   Menacho-Melgar R, Ye Z, Moreb EA, Yang T, Efromson JP, Decker JS, 
            Wang R, Lynch MD
  TITLE     Scalable, two-stage, autoinduction of recombinant protein expression
            in E. coli utilizing phosphate depletion.
  JOURNAL   Biotechnol Bioeng. 2020 May 22. doi: 10.1002/bit.27440.
  PUBMED    32441815
REFERENCE   2  (bases 1 to 2981)
  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: "Biotechnol 
            Bioeng. 2020 May 22. doi: 10.1002/bit.27440."
COMMENT     Sequence Label: pHCKan-INSyibp-Nef
FEATURES             Location/Qualifiers
     source          1..2981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      65..96
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        97..199
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             200..1015
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(273..292)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1036..1063
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1075..1662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1155..1174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1684..1713)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      1797..1868
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1884..1911
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2173..2184
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2821..2844
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2845..2862
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gacgaattct ctagatatcg ctcaatactg accatttaaa tcatacctga cctccatagc
       61 agaaagtcaa aagcctccga ccggaggctt ttgacttgat cggcacgtaa gaggttccaa
      121 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
      181 aggagctaag gaagctaaaa tgagccatat tcaacgggaa acgtcttgct cgaggccgcg
      241 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
      301 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
      361 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca ggctaaactg
      421 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
      481 atggttactc accactgcga tcccagggaa aacagcattc caggtattag aagaatatcc
      541 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
      601 tcctgtttgt aattgtcctt ttaacggcga tcgcgtattt cgtctcgctc aggcgcaatc
      661 acgaatgaat aacggtttgg ttggtgcgag tgattttgat gacgagcgta atggctggcc
      721 tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt
      781 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
      841 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
      901 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
      961 taatcctgat atgaataaat tgcagtttca cttgatgctc gatgagtttt tctgagggcc
     1021 caaatgtaat cacctggctc accttcgggt gggcctttct gcgttgctgg cgtttttcca
     1081 taggctccgc ccccctgacg agcatcacaa aaatcgatgc tcaagtcaga ggtggcgaaa
     1141 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1201 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1261 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1321 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1381 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1441 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1501 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttacctcgga
     1561 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1621 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgattttc
     1681 taccgaagaa aggcccaccc gtgaaggtga gccagtgagt tgattgcagt ccagttacgc
     1741 tggagtctga ggctcgtcct gaatgatatc aagcttgaat tcgtttgccc aggcatcaaa
     1801 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     1861 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatacac
     1921 agctaacacc acgtcgtccc tatctgctgc cctaggtcta tgagtggttg ctggataacg
     1981 tgcgtaattg tgctgatctc ttatatagct gctctcatta tctctctacc ctgaagtgac
     2041 tctctcacct gtaaaaataa tatctcacag gcttaatagt ttcttaatac aaagcctgta
     2101 aaacgtcagg ataacttcta tattcaggga gaccacaacg gtttccctct acaaataatt
     2161 ttgtttaact ttaaagagga gaaatactag atgggtggca agtggtcaaa acgtagtatg
     2221 ggtggatggt ctgctataag ggaaagaatg agacgagctg agccacgagc tgagccagca
     2281 gcagatgggg tgggagcagt atctcgagac ctggaaaaac atggagcaat cacaagtagc
     2341 aatacagcag ctactaatgc tgattgtgcc tggctagaag cacaagagga ggaagaggtg
     2401 ggttttccag tcagacctca ggtaccttta agaccaatga cttacaaggc agctttagat
     2461 attagccact ttttaaaaga aaagggggga ctggaagggc taatttggtc ccaaagaaga
     2521 caagagatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaat
     2581 tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgctt caagctagta
     2641 ccagttgagc cagagaaggt agaagaggcc aatgaaggag agaacaacag cttgttacac
     2701 cctatgagcc tgcatgggat ggaggacgcg gagaaagaag tgttagtgtg gaggtttgac
     2761 agcaaactag catttcatca catggcccga gagctgcatc cggagtacta caaagactgc
     2821 ctcgaagttc tgttccaggg gccccatcac catcatcacc actaatgacc ggcttatcgg
     2881 tcagtttcac ctgatttacg taaaaacccg cttcggcggg tttttgcttt tggaggggca
     2941 gaaagatgaa tgactgtcca cgacgctata cccaaaagaa a
//
