LOCUS       sequence-119580-        3451 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 3451)
  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 3451)
  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-Ala1
FEATURES             Location/Qualifiers
     source          1..3451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      9..80
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      96..123
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     1528..1547
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1528..1546
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1725..1756
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1757..1859
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1860..2675
                     /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(1933..1952)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2699..2726
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2738..3325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2818..2837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3347..3376)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
       61 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
      121 gcgtttatac acagctaaca ccacgtcgtc cctatctgct gccctaggtc tatgagtggt
      181 tgctggataa ctctttctga caccttacta tcttacaaat gtaacaaaaa agttattttt
      241 ctgtaattcg agcatgtcat gttaccccgc gagcataaaa cgcgtatatt cagggagacc
      301 acaacggttt ccctctacaa ataattttgt ttaactttcg tagaagagca cttcacactt
      361 ctggaaaaag gagatatacc atgatcattg gggtgccgaa ggagatcaaa aataatgaga
      421 accgcgtcgc gttgaccccg ggaggtgtca gccagctgat ctctaatggc catcgtgtct
      481 tagttgaaac aggcgctggc ctgggttctg gcttcgaaaa cgaggcctac gaatctgcag
      541 gtgcggaaat tattgctgat ccaaaacagg tctgggatgc agagatggtc atgaaagtga
      601 aagaaccgct cccggaagaa tatgtctatt ttcgtaaagg tctggtgctg tttacatatc
      661 tgcatctggc agctgaaccg gagctcgcac aagcccttaa agataaaggt gtcacggcca
      721 tcgcatacga aactgtcagc gaagggcgca cgctgccatt actgaccccg atgtcagaag
      781 tggcaggccg tatggctgcg cagatcggcg cacagtttct tgaaaaacca aagggcggga
      841 agggtattct cttagcagga gtgccgggcg tcagtcgtgg gaaagtaact attattggtg
      901 gcggcgtggt aggaacaaat gctgccaaaa tggccgtcgg tttgggggcc gacgtaacaa
      961 tcattgcgcg taatgccgat cgccttcgtc aattagacga tatctttggc caccaaatca
     1021 aaaccctgat ttcgaaccca gtcaatatcg cggatgcggt ggcggaagct gatttgttga
     1081 tctgcgccgt gttaattccg ggagcgaaag cacctacatt ggtgacggaa gaaatggtga
     1141 aacaaatgaa accgggttca gtcattgttg atgtggctat tgatcagggt ggcatcgtgg
     1201 aaacggtgga ccatattacc actcacgacc agccgacgta tgaaaaacat ggtgtcgtac
     1261 actatgcggt ggcgaatatg cctggtgcgg tcccacgtac gagtacaatc gcactgacaa
     1321 atgtcaccgt gccgtatgcg ttgcaaatcg cgaacaaagg tgccgtgaaa gcgctggccg
     1381 acaatacggc gttacgtgcc ggtctgaaca ccgctaacgg tcacgtgaca tatgaagcgg
     1441 tcgcgcgtga tttggggtac gaatatgtac cggcggaaaa agccttacaa gacgaatcga
     1501 gtgtcgctgg tgcatagtaa gctcttctaa tacgactcac tatagggccg gcttatcggt
     1561 cagtttcacc tgatttacgt aaaaacccgc ttcggcgggt ttttgctttt ggaggggcag
     1621 aaagatgaat gactgtccac gacgctatac ccaaaagaaa gacgaattct ctagatatcg
     1681 ctcaatactg accatttaaa tcatacctga cctccatagc agaaagtcaa aagcctccga
     1741 ccggaggctt ttgacttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa
     1801 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa
     1861 tgagccatat tcaacgggaa acgtcttgct cgaggccgcg attaaattcc aacatggatg
     1921 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     1981 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     2041 ttgccaatga tgttacagat gagatggtca ggctaaactg gctgacggaa tttatgcctc
     2101 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     2161 tcccagggaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     2221 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     2281 ttaacggcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     2341 ttggtgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     2401 aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     2461 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     2521 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     2581 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     2641 tgcagtttca cttgatgctc gatgagtttt tctaatgagg gcccaaatgt aatcacctgg
     2701 ctcaccttcg ggtgggcctt tctgcgttgc tggcgttttt ccataggctc cgcccccctg
     2761 acgagcatca caaaaatcga tgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2821 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2881 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2941 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3001 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3061 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3121 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3181 cagtatttgg tatctgcgct ctgctgaagc cagttacctc ggaaaaagag ttggtagctc
     3241 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3301 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatt ttctaccgaa gaaaggccca
     3361 ccagtgaagg tgagccagtg agttgattgc agtccagtta cgctggagtc tgaggctcgt
     3421 cctgaatgat atcaagcttg aattcgtttg c
//
