LOCUS       sequence-271207-        3557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low Phosphate inducible mutant citramalate synthase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3557)
  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 3557)
  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-yibDp-cimA3.7
FEATURES             Location/Qualifiers
     source          1..3557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1449..1465)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1449..1465)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1462..1484)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1637..2225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1717..1736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2361..3176)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2361..3176)
                     /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
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3084..3103
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3168..3187
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3297..3313
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3297..3313
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      3314..3360
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3474..3496
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3488..3505
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3489..3505
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtgcgtaatt gtgctgatct cttatatagc tgctctcatt atctctctac cctgaagtga
       61 ctctctcacc tgtaaaaata atatctcaca ggcttaatag tttcttaata caaagcctgt
      121 aaaacgtcag gataacttct gtgtaggagg ataatctatg atggtccgta tttttgatac
      181 gactttgcgt gatggggagc agacacctgg cgtctccctt acaccgaacg acaagttaga
      241 gattgctaaa aagttagatg agcttggggt agacgttatc gaggcgggct cagcggtcac
      301 tagcaaggga gagcgcgagg ggatcaagtt gattaccaag gaaggcctta atgcggagat
      361 ttgttcgttc gtccgcgccc tgcccgtaga catcgatgcc gcacttgagt gcgatgtgga
      421 ttcagtacat ttagtcgtac caacttcccc gatccatatg aaatacaagt tgcgtaaaac
      481 cgaggatgaa gtccttgtca cagcgctgaa agcagtggag tacgcgaagg agcaaggctt
      541 gatcgtcgaa ttgtcagccg aagatgcaac ccgctcggac gtgaactttc ttatcaagct
      601 tttcaacgaa ggagagaaag ttggagcgga tcgtgtgtgt gtatgtgata cagtcggcgt
      661 tttgacacca caaaagtcac aggagctgtt caagaaaatc acggagaacg tgaacctgcc
      721 cgtgtcagta cattgccata acgacttcgg catggccacg gccaacgcgt gttcagcagt
      781 gcttggtgga gcagttcaat gccacgttac ggttaatggt attggcgaac gcgcgggaaa
      841 tgcttcatta gaggaagtcg tcgccgcgtc aaaaatcttg tacggctacg acaccaaaat
      901 caagatggag aaactttacg aggtgtcgcg tattgtatcc cgtctgatga aacttcctgt
      961 accccccaac aaggcgatcg taggagataa cgccttcgcg cacgaggctg ggattcacgt
     1021 ggacggactg attaaaaaca ctgagactta tgagcccatc aaaccggaaa tggtcgggaa
     1081 ccgccgccgt atcattttag gaaaacattc gggacgtaag gcgctgaaat acaaattaga
     1141 cctgatgggc atcaatgtct ccgacgagca attaaacaaa atctatgaac gtgtgaagga
     1201 attcggtgac ttagggaagt atatctccga cgccgattta ctggcgattg tgcgcgaagt
     1261 aacggggaag ttagtataac cggcttatcg gtcagtttca cctgatttac gtaaaaaccc
     1321 gcttcggcgg gtttttgctt ttggaggggc agaaagatga atgactgtcc acgacgctat
     1381 acccaaaaga aaaggctagg tggaggctca gtgatgataa gtctgcgatg gtggatgcat
     1441 gtgtcatggt catagctgtt tcctgtgtga aattgttatc cgctcagagg gcacaatcct
     1501 attccgcgct atccgacaat ctccaagaca ttaggtggag ttcagttcgg cgtatggcat
     1561 atgtcgctgg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1621 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1681 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1741 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1801 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1861 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1921 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1981 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2041 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2101 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2161 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2221 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tctattcaac aaagccgccg
     2281 tcccgtcaag tcagcgtaaa tgggtagggg gcttcaaatc gtccgctctg ccagtgttac
     2341 aaccaattaa caaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     2401 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     2461 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     2521 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     2581 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
     2641 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     2701 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     2761 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     2821 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     2881 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     2941 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     3001 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     3061 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     3121 atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg gctcataaca
     3181 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     3241 tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttcccc cgccgctcta
     3301 gaactagtgg atccaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     3361 ctgttgtttg tcgcattata cgagacgtcc aggttgggat acctgaaaca aaacccatcg
     3421 tacggccaag gaagtctcca ataactgtga tccaccacaa gcgccagggt tttcccagtc
     3481 acgacgttgt aaaacgacgg ccagtcatgc ataatccgca cgcatctgga ataaggaagt
     3541 gccattccgc ctgacct
//
