LOCUS       sequence-174932-        3319 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 3319)
  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 3319)
  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-ala11
FEATURES             Location/Qualifiers
     source          1..3319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1451..1470
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1451..1469
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1648..1679
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1680..1782
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1783..2598
                     /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(1856..1875)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2619..2646
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2658..3245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2738..2757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3267..3296)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgttt gccgccgaaa
       61 accccgcttc ggcggggttt tgccgcacgt ctccatcgct tgcccaagtt gtgaagcaca
      121 gctaacacca cgtcgtccct atctgctgcc ctaggtctat gagtggttgc tggataactt
      181 tacgggcatg cataaggctc gtaggctata ttcagggaga ccacaacggt ttccctctac
      241 aaataatttt gtttaacttt cgtagaagag cacttccaca cttctggaaa aaggagatat
      301 accatgatca ttggggtgcc gaaggagatc aaaaataatg agaaccgcgt cgcgttgacc
      361 ccgggaggtg tcagccagct gatctctaat ggccatcgtg tcttagttga aacaggcgct
      421 ggcctgggtt ctggcttcga aaacgaggcc tacgaatctg caggtgcgga aattattgct
      481 gatccaaaac aggtctggga tgcagagatg gtcatgaaag tgaaagaacc gctcccggaa
      541 gaatatgtct attttcgtaa aggtctggtg ctgtttacat atctgcatct ggcagctgaa
      601 ccggagctcg cacaagccct taaagataaa ggtgtcacgg ccatcgcata cgaaactgtc
      661 agcgaagggc gcacgctgcc attactgacc ccgatgtcag aagtggcagg ccgtatggct
      721 gcgcagatcg gcgcacagtt tcttgaaaaa ccaaagggcg ggaagggtat tctcttagca
      781 ggagtgccgg gcgtcagtcg tgggaaagta actattattg gtggcggcgt ggtaggaaca
      841 aatgctgcca aaatggccgt cggtttgggg gccgacgtaa caatcattgc gcgtaatgcc
      901 gatcgccttc gtcaattaga cgatatcttt ggccaccaaa tcaaaaccct gatttcgaac
      961 ccagtcaata tcgcggatgc ggtggcggaa gctgatttgt tgatctgcgc cgtgttaatt
     1021 ccgggagcga aagcacctac attggtgacg gaagaaatgg tgaaacaaat gaaaccgggt
     1081 tcagtcattg ttgatgtggc tattgatcag ggtggcatcg tggaaacggt ggaccatatt
     1141 accactcacg accagccgac gtatgaaaaa catggtgtcg tacactatgc ggtggcgaat
     1201 atgcctggtg cggtcccacg tacgagtaca atcgcactga caaatgtcac cgtgccgtat
     1261 gcgttgcaaa tcgcgaacaa aggtgccgtg aaagcgctgg ccgacaatac ggcgttacgt
     1321 gccggtctga acaccgctaa cggtcacgtg acatatgaag cggtcgcgcg tgatttgggg
     1381 tacgaatatg taccggcgga aaaagcctta caagacgaat cgagtgtcgc tggtgcatag
     1441 taagctcttc taatacgact cactataggg ccggcttatc ggtcagtttc acctgattta
     1501 cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg cagaaagatg aatgactgtc
     1561 cacgacgcta tacccaaaag aaagacgaat tctctagata tcgctcaata ctgaccattt
     1621 aaatcatacc tgacctccat agcagaaagt caaaagcctc cgaccggagg cttttgactt
     1681 gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact accgggcgta
     1741 ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatgagcca tattcaacgg
     1801 gaaacgtctt gctcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     1861 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag
     1921 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     1981 gatgagatgg tcaggctaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     2041 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatcccagg gaaaacagca
     2101 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     2161 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacgg cgatcgcgta
     2221 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttggtgc gagtgatttt
     2281 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg
     2341 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     2401 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     2461 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     2521 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcacttgatg
     2581 ctcgatgagt ttttctgagg gcccaaatgt aatcacctgg ctcaccttcg ggtgggcctt
     2641 tctgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2701 tgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2761 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2821 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2881 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2941 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3001 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3061 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3121 ctgctgaagc cagttacctc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3181 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3241 ctcaagaaga tcctttgatt ttctaccgaa gaaaggccca cccgtgaagg tgagccagtg
     3301 agttgattgc agtccagtt
//
