LOCUS       sequence-132780-        3497 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 3497)
  AUTHORS   Ye Z, Li S, Hennigan JN, Lebeau J, Moreb EA, Wolf J, Lynch MD
  TITLE     Two-stage dynamic deregulation of metabolism improves process 
  JOURNAL   Metab Eng. 2021 Nov;68:106-118. doi: 10.1016/j.ymben.2021.09.009. 
            Epub 2021 Sep 30.
  PUBMED    34600151
REFERENCE   2  (bases 1 to 3497)
  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: "Metab 
            Eng."; date: "2021-11"; volume: "68:106-118. doi"; pages: " 
            10.1016/j.ymben.2021.09.009. Epub 2021 Sep 30"
COMMENT     Sequence Label: pSMART-Ala2
FEATURES             Location/Qualifiers
     source          1..3497
                     /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     1574..1593
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1574..1592
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1771..1802
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1803..1905
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1906..2721
                     /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(1979..1998)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2745..2772
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2784..3371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2864..2883)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3393..3422)
                     /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 cgtgcgtaat tgtgctgatc tcttatatag ctgctctcat tatctctcta
      241 ccctgaagtg actctctcac ctgtaaaaat aatatctcac aggcttaata gtttcttaat
      301 acaaagcctg taaaacgtca ggataacttc tatattcagg gagaccacaa cggtttccct
      361 ctacaaataa ttttgtttaa ctttcgtaga agagcacttc acacttctgg aaaaaggaga
      421 tataccatga tcattggggt gccgaaggag atcaaaaata atgagaaccg cgtcgcgttg
      481 accccgggag gtgtcagcca gctgatctct aatggccatc gtgtcttagt tgaaacaggc
      541 gctggcctgg gttctggctt cgaaaacgag gcctacgaat ctgcaggtgc ggaaattatt
      601 gctgatccaa aacaggtctg ggatgcagag atggtcatga aagtgaaaga accgctcccg
      661 gaagaatatg tctattttcg taaaggtctg gtgctgttta catatctgca tctggcagct
      721 gaaccggagc tcgcacaagc ccttaaagat aaaggtgtca cggccatcgc atacgaaact
      781 gtcagcgaag ggcgcacgct gccattactg accccgatgt cagaagtggc aggccgtatg
      841 gctgcgcaga tcggcgcaca gtttcttgaa aaaccaaagg gcgggaaggg tattctctta
      901 gcaggagtgc cgggcgtcag tcgtgggaaa gtaactatta ttggtggcgg cgtggtagga
      961 acaaatgctg ccaaaatggc cgtcggtttg ggggccgacg taacaatcat tgcgcgtaat
     1021 gccgatcgcc ttcgtcaatt agacgatatc tttggccacc aaatcaaaac cctgatttcg
     1081 aacccagtca atatcgcgga tgcggtggcg gaagctgatt tgttgatctg cgccgtgtta
     1141 attccgggag cgaaagcacc tacattggtg acggaagaaa tggtgaaaca aatgaaaccg
     1201 ggttcagtca ttgttgatgt ggctattgat cagggtggca tcgtggaaac ggtggaccat
     1261 attaccactc acgaccagcc gacgtatgaa aaacatggtg tcgtacacta tgcggtggcg
     1321 aatatgcctg gtgcggtccc acgtacgagt acaatcgcac tgacaaatgt caccgtgccg
     1381 tatgcgttgc aaatcgcgaa caaaggtgcc gtgaaagcgc tggccgacaa tacggcgtta
     1441 cgtgccggtc tgaacaccgc taacggtcac gtgacatatg aagcggtcgc gcgtgatttg
     1501 gggtacgaat atgtaccggc ggaaaaagcc ttacaagacg aatcgagtgt cgctggtgca
     1561 tagtaagctc ttctaatacg actcactata gggccggctt atcggtcagt ttcacctgat
     1621 ttacgtaaaa acccgcttcg gcgggttttt gcttttggag gggcagaaag atgaatgact
     1681 gtccacgacg ctatacccaa aagaaagacg aattctctag atatcgctca atactgacca
     1741 tttaaatcat acctgacctc catagcagaa agtcaaaagc ctccgaccgg aggcttttga
     1801 cttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
     1861 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgag ccatattcaa
     1921 cgggaaacgt cttgctcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     1981 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg
     2041 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     2101 acagatgaga tggtcaggct aaactggctg acggaattta tgcctcttcc gaccatcaag
     2161 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc agggaaaaca
     2221 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     2281 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cggcgatcgc
     2341 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttgg tgcgagtgat
     2401 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt
     2461 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2521 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2581 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2641 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcacttg
     2701 atgctcgatg agtttttcta atgagggccc aaatgtaatc acctggctca ccttcgggtg
     2761 ggcctttctg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2821 aatcgatgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2881 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2941 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3001 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3061 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3121 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3181 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3241 tgcgctctgc tgaagccagt tacctcggaa aaagagttgg tagctcttga tccggcaaac
     3301 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3361 aaggatctca agaagatcct ttgattttct accgaagaaa ggcccacccg tgaaggtgag
     3421 ccagtgagtt gattgcagtc cagttacgct ggagtctgag gctcgtcctg aatgatatca
     3481 agcttgaatt cgtttgc
//
