LOCUS       sequence-174449-        3324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NADPH dependent Alanine Dehydrogenase, Constitutive E. coli 
            Expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3324)
  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 3324)
  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-ala12
FEATURES             Location/Qualifiers
     source          1..3324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1404..1423
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1404..1422
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1601..1632
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1633..1735
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1736..2551
                     /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(1809..1828)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2572..2599
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2611..3198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2691..2710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3220..3249)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cgccgcaaac cccgcccctg acagggcggg gtttcgccgc acgtctccat cgcttgccca
       61 agttgtgaag cacagctaac accacgtcgt ccctatctgc tgccctaggt ctatgagtgg
      121 ttgctggata actttacggg catgcataag gctcgtatga tatattcagg gagaccacaa
      181 cggtttccct ctacaaataa ttttgtttaa ctttcgtaga gagcacttcc acacttctgg
      241 aaaaaggaga tataccatga tcattggggt gccgaaggag atcaaaaata atgagaaccg
      301 cgtcgcgttg accccgggag gtgtcagcca gctgatctct aatggccatc gtgtcttagt
      361 tgaaacaggc gctggcctgg gttctggctt cgaaaacgag gcctacgaat ctgcaggtgc
      421 ggaaattatt gctgatccaa aacaggtctg ggatgcagag atggtcatga aagtgaaaga
      481 accgctcccg gaagaatatg tctattttcg taaaggtctg gtgctgttta catatctgca
      541 tctggcagct gaaccggagc tcgcacaagc ccttaaagat aaaggtgtca cggccatcgc
      601 atacgaaact gtcagcgaag ggcgcacgct gccattactg accccgatgt cagaagtggc
      661 aggccgtatg gctgcgcaga tcggcgcaca gtttcttgaa aaaccaaagg gcgggaaggg
      721 tattctctta gcaggagtgc cgggcgtcag tcgtgggaaa gtaactatta ttggtggcgg
      781 cgtggtagga acaaatgctg ccaaaatggc cgtcggtttg ggggccgacg taacaatcat
      841 tgcgcgtaat gccgatcgcc ttcgtcaatt agacgatatc tttggccacc aaatcaaaac
      901 cctgatttcg aacccagtca atatcgcgga tgcggtggcg gaagctgatt tgttgatctg
      961 cgccgtgtta attccgggag cgaaagcacc tacattggtg acggaagaaa tggtgaaaca
     1021 aatgaaaccg ggttcagtca ttgttgatgt ggctattgat cagggtggca tcgtggaaac
     1081 ggtggaccat attaccactc acgaccagcc gacgtatgaa aaacatggtg tcgtacacta
     1141 tgcggtggcg aatatgcctg gtgcggtccc acgtacgagt acaatcgcac tgacaaatgt
     1201 caccgtgccg tatgcgttgc aaatcgcgaa caaaggtgcc gtgaaagcgc tggccgacaa
     1261 tacggcgtta cgtgccggtc tgaacaccgc taacggtcac gtgacatatg aagcggtcgc
     1321 gcgtgatttg gggtacgaat atgtaccggc ggaaaaagcc ttacaagacg aatcgagtgt
     1381 cgctggtgca tagtaagctc ttctaatacg actcactata gggccggctt atcggtcagt
     1441 ttcacctgat ttacgtaaaa acccgcttcg gcgggttttt gcttttggag gggcagaaag
     1501 atgaatgact gtccacgacg ctatacccaa aagaaagacg aattctctag atatcgctca
     1561 atactgacca tttaaatcat acctgacctc catagcagaa agtcaaaagc ctccgaccgg
     1621 aggcttttga cttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
     1681 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgag
     1741 ccatattcaa cgggaaacgt cttgctcgag gccgcgatta aattccaaca tggatgctga
     1801 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1861 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1921 caatgatgtt acagatgaga tggtcaggct aaactggctg acggaattta tgcctcttcc
     1981 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2041 agggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2101 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2161 cggcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttgg
     2221 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2281 gcataagctt ttgccattct caccggattc agtcgtcacc catggtgatt tctcacttga
     2341 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2401 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2461 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2521 gtttcacttg atgctcgatg agtttttctg agggcccaaa tgtaatcacc tggctcacct
     2581 tcgggtgggc ctttctgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2641 tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2701 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2761 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2821 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2881 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2941 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3001 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3061 tggtatctgc gctctgctga agccagttac ctcggaaaaa gagttggtag ctcttgatcc
     3121 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3181 agaaaaaaag gatctcaaga agatcctttg attttctacc gaagaaaggc ccacccgtga
     3241 aggtgagcca gtgagttgat tgcagtccag ttacgctgga gtctgaggct cgtcctgaat
     3301 gatatcaagc ttgaattcgt ttgc
//
