LOCUS       sequence-174451-        3962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NADPH dependent Alanine Dehydrogenase, Low Phosphate Dependent E. 
            coli Expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3962)
  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 3962)
  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: pSMART-ala10
FEATURES             Location/Qualifiers
     source          1..3962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      50..121
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      137..164
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     2083..2102
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2083..2101
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2280..2311
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2312..2414
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2415..3230
                     /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(2488..2507)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3251..3278
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3290..3877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3370..3389)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3899..3928)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 tgaggctcgt cctgaatgat atcaagcttg aattcgtttg cccaggcatc aaataaaacg
       61 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
      121 ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata cacagctaac
      181 accacgtcgt ccctatctgc tgccctaggt ctatgagtgg ttgctggata acgtgcgtaa
      241 ttgtgctgat ctcttatata gctgctctca ttatctctct accctgaagt gactctctca
      301 cctgtaaaaa taatatctca caggcttaat agtttcttaa tacaaagcct gtaaaacgtc
      361 aggataactt ctatattcag ggagaccaca acggtttccc tctacaaata attttgttta
      421 actttcgtag aagagcactt cacacttctg gaaaaaggag atataccatg atcattgggg
      481 tgccgaagga gatcaaaaat aatgagaacc gcgtcgcgtt gaccccggga ggtgtcagcc
      541 agctgatctc taatggccat cgtgtcttag ttgaaacagg cgctggcctg ggttctggct
      601 tcgaaaacga ggcctacgaa tctgcaggtg cggaaattat tgctgatcca aaacaggtct
      661 gggatgcaga gatggtcatg aaagtgaaag aaccgctccc ggaagaatat gtctattttc
      721 gtaaaggtct ggtgctgttt acatatctgc atctggcagc tgaaccggag ctcgcacaag
      781 cccttaaaga taaaggtgtc acggccatcg catacgaaac tgtcagcgaa gggcgcacgc
      841 tgccattact gaccccgatg tcagaagtgg caggccgtat ggctgcgcag atcggcgcac
      901 agtttcttga aaaaccaaag ggcgggaagg gtattctctt agcaggagtg ccgggcgtca
      961 gtcgtgggaa agtaactatt attggtggcg gcgtggtagg aacaaatgct gccaaaatgg
     1021 ccgtcggttt gggggccgac gtaacaatca ttgcgcgtaa tgccgatcgc cttcgtcaat
     1081 tagacgatat ctttggccac caaatcaaaa ccctgatttc gaacccagtc aatatcgcgg
     1141 atgcggtggc ggaagctgat ttgttgatct gcgccgtgtt aattccggga gcgaaagcac
     1201 ctacattggt gacggaagaa atggtgaaac aaatgaaacc gggttcagtc attgttgatg
     1261 tggctattga tcagggtggc atcgtggaaa cggtggacca tattaccact cacgaccagc
     1321 cgacgtatga aaaacatggt gtcgtacact atgcggtggc gaatatgcct ggtgcggtcc
     1381 cacgtacgag tacaatcgca ctgacaaatg tcaccgtgcc gtatgcgttg caaatcgcga
     1441 acaaaggtgc cgtgaaagcg ctggccgaca atacggcgtt acgtgccggt ctgaacaccg
     1501 ctaacggtca cgtgacatat gaagcggtcg cgcgtgattt ggggtacgaa tatgtaccgg
     1561 cggaaaaagc cttacaagac gaatcgagtg tcgctggtgc atagtaagga aaaaggagat
     1621 ataccatgtt ctcaccgcag tcacgcttgc gtcatgcagt tgcagatacg ttcgcgatgg
     1681 ttgtttactg ttctgtcgtg aacatgtgta ttgaagtttt cctctccgga atgagcttcg
     1741 aacagtcttt ttattccaga ttggtagcga ttccggtgaa catcttaatt gcatggccat
     1801 acggtatgta ccgtgatctg tttatgcgcg cggcacgcaa agttagcccg tcgggctgga
     1861 taaaaaatct ggcggatatc ctggcttatg tgacgttcca gtcaccggtg tatgtggcga
     1921 tcttgttagt ggtgggcgca gactggcatc agattatggc ggcggtcagt tcaaacatcg
     1981 ttgtttcgat gttgatgggg gcggtttatg gctacttcct cgattattgc cgccgactgt
     2041 ttaaagtcag ccgttaccag caggtaaaag cctgagctct tctaatacga ctcactatag
     2101 ggccggctta tcggtcagtt tcacctgatt tacgtaaaaa cccgcttcgg cgggtttttg
     2161 cttttggagg ggcagaaaga tgaatgactg tccacgacgc tatacccaaa agaaagacga
     2221 attctctaga tatcgctcaa tactgaccat ttaaatcata cctgacctcc atagcagaaa
     2281 gtcaaaagcc tccgaccgga ggcttttgac ttgatcggca cgtaagaggt tccaactttc
     2341 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
     2401 ctaaggaagc taaaatgagc catattcaac gggaaacgtc ttgctcgagg ccgcgattaa
     2461 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     2521 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac
     2581 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcaggcta aactggctga
     2641 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     2701 tactcaccac tgcgatccca gggaaaacag cattccaggt attagaagaa tatcctgatt
     2761 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     2821 tttgtaattg tccttttaac ggcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     2881 tgaataacgg tttggttggt gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     2941 aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc
     3001 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     3061 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     3121 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     3181 ctgatatgaa taaattgcag tttcacttga tgctcgatga gtttttctga gggcccaaat
     3241 gtaatcacct ggctcacctt cgggtgggcc tttctgcgtt gctggcgttt ttccataggc
     3301 tccgcccccc tgacgagcat cacaaaaatc gatgctcaag tcagaggtgg cgaaacccga
     3361 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3421 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3481 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3541 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3601 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3661 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3721 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc tcggaaaaag
     3781 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3841 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga ttttctaccg
     3901 aagaaaggcc cacccgtgaa ggtgagccag tgagttgatt gcagtccagt tacgctggag
     3961 tc
//
