LOCUS       sequence-180934-        3318 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 3318)
  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 3318)
  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-ala13
FEATURES             Location/Qualifiers
     source          1..3318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1398..1417
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1398..1416
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1595..1626
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1627..1729
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1730..2545
                     /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(1803..1822)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2566..2593
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2605..3192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2685..2704)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3214..3243)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cgcaaaaaac cccgcttcgg cggggttttt tcgcacgtct ccatcgcttg cccaagttgt
       61 gaagcacagc taacaccacg tcgtccctat ctgctgccct aggtctatga gtggttgctg
      121 gataacttta cgggcatgca taaggctcgt ataatatatt cagggagacc acaacggttt
      181 ccctctacaa ataattttgt ttaactttcg tagaagagca cttccacact tctggaaaaa
      241 gagatatacc atgatcattg gggtgccgaa ggagatcaaa aataatgaga accgcgtcgc
      301 gttgaccccg ggaggtgtca gccagctgat ctctaatggc catcgtgtct tagttgaaac
      361 aggcgctggc ctgggttctg gcttcgaaaa cgaggcctac gaatctgcag gtgcggaaat
      421 tattgctgat ccaaaacagg tctgggatgc agagatggtc atgaaagtga aagaaccgct
      481 cccggaagaa tatgtctatt ttcgtaaagg tctggtgctg tttacatatc tgcatctggc
      541 agctgaaccg gagctcgcac aagcccttaa agataaaggt gtcacggcca tcgcatacga
      601 aactgtcagc gaagggcgca cgctgccatt actgaccccg atgtcagaag tggcaggccg
      661 tatggctgcg cagatcggcg cacagtttct tgaaaaacca aagggcggga agggtattct
      721 cttagcagga gtgccgggcg tcagtcgtgg gaaagtaact attattggtg gcggcgtggt
      781 aggaacaaat gctgccaaaa tggccgtcgg tttgggggcc gacgtaacaa tcattgcgcg
      841 taatgccgat cgccttcgtc aattagacga tatctttggc caccaaatca aaaccctgat
      901 ttcgaaccca gtcaatatcg cggatgcggt ggcggaagct gatttgttga tctgcgccgt
      961 gttaattccg ggagcgaaag cacctacatt ggtgacggaa gaaatggtga aacaaatgaa
     1021 accgggttca gtcattgttg atgtggctat tgatcagggt ggcatcgtgg aaacggtgga
     1081 ccatattacc actcacgacc agccgacgta tgaaaaacat ggtgtcgtac actatgcggt
     1141 ggcgaatatg cctggtgcgg tcccacgtac gagtacaatc gcactgacaa atgtcaccgt
     1201 gccgtatgcg ttgcaaatcg cgaacaaagg tgccgtgaaa gcgctggccg acaatacggc
     1261 gttacgtgcc ggtctgaaca ccgctaacgg tcacgtgaca tatgaagcgg tcgcgcgtga
     1321 tttggggtac gaatatgtac cggcggaaaa agccttacaa gacgaatcga gtgtcgctgg
     1381 tgcatagtaa gctcttctaa tacgactcac tatagggccg gcttatcggt cagtttcacc
     1441 tgatttacgt aaaaacccgc ttcggcgggt ttttgctttt ggaggggcag aaagatgaat
     1501 gactgtccac gacgctatac ccaaaagaaa gacgaattct ctagatatcg ctcaatactg
     1561 accatttaaa tcatacctga cctccatagc agaaagtcaa aagcctccga ccggaggctt
     1621 ttgacttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     1681 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tgagccatat
     1741 tcaacgggaa acgtcttgct cgaggccgcg attaaattcc aacatggatg ctgatttata
     1801 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta
     1861 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     1921 tgttacagat gagatggtca ggctaaactg gctgacggaa tttatgcctc ttccgaccat
     1981 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tcccagggaa
     2041 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2101 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacggcga
     2161 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttggtgcgag
     2221 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2281 gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2341 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2401 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2461 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2521 cttgatgctc gatgagtttt tctgagggcc caaatgtaat cacctggctc accttcgggt
     2581 gggcctttct gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2641 aaatcgatgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2701 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2761 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2821 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2881 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2941 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3001 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3061 ctgcgctctg ctgaagccag ttacctcgga aaaagagttg gtagctcttg atccggcaaa
     3121 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3181 aaaggatctc aagaagatcc tttgattttc taccgaagaa aggcccaccc gtgaaggtga
     3241 gccagtgagt tgattgcagt ccagttacgc tggagtctga ggctcgtcct gaatgatatc
     3301 aagcttgaat tcgtttgc
//
