LOCUS       sequence-334102-        4954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PgyrA_ada _adh2 _pMB1_ampR (for expressing two enzymes (Ada and 
            Adh2) to allow E. coli to consume ethanol).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4954)
  AUTHORS   Liang H, Ma X, Ning W, Liu Y, Sinskey AJ, Stephanopoulos G, Zhou K
  TITLE     Constructing an ethanol utilization pathway in Escherichia coli to 
            produce acetyl-CoA derived compounds.
  JOURNAL   Metab Eng. 2020 Nov 25. pii: S1096-7176(20)30180-4. doi: 
            10.1016/j.ymben.2020.11.010.
  PUBMED    33248272
REFERENCE   2  (bases 1 to 4954)
  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. 
            2020 Nov 25. pii: S1096-7176(20)30180-4. doi: 
            10.1016/j.ymben.2020.11.010."
COMMENT     Sequence Label: Plasmid EUPp_09
FEATURES             Location/Qualifiers
     source          1..4954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2549..2567)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2563..2610
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2740..2831
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2832..2900,2901..3692)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3050..3069)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3882..4470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4371..4390
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4624..4641
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             4721..4738
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tttgtttaac tttaagaagg agatatacat atggaacact ccgtgatcga accgactgta
       61 ccgatgccca tgccagcgat gtttgacgca ccgagcggca ttttcgactc gcttgacgac
      121 gccgttcagg ccgccgtgta cgcgcaacaa cagttgaaca gcgtcgagct gcgtcaacag
      181 gtgattaaag ccattcgggt cgctggcgag cgttatgccc aggtgctggc ggaaatggcc
      241 gtggccgaaa ccggcatggg ccgcgtggtc gataagtaca tcaaaaacgt gtcacaggcg
      301 cgccatacac cagggatcga atgcctgtcg gcggaagtac tgaccggtga taatgggctg
      361 acgctgattg aaaatgcccc ctggggtgtg gttgcgtcgg tcaccccgtc aaccaacccg
      421 gccgcaaccg ttatcaataa cgccatcagt atgatagcag ccggtaacag cgtggtgttc
      481 gcccctcatc cgtcagcgaa aaacgtatcg ctgcgtacca tcagcctgct caataaggcc
      541 atcgtcgcca ccggcggccc ggagaacctg ctggtcagcg tttccgaccc caacatcgaa
      601 accgcgcaac gcctgtttcg ctaccccggt atcgggctgc tggttgtcac cggcggcgaa
      661 gcggtggtgg aagcggcgcg cagacatacc gacaaacggc tgatcgccgc cggagcgggt
      721 aacccgccag tggtggtgga tgaaacggcg gatattccga aagcagcccg cgcgattgtt
      781 aaaggcgcct cattcgacaa caacattatc tgtgccgatg agaaggtgct gatcgtggtg
      841 gacagtgtgg ccgatgccct actggcagaa atgcaacgca atcatgccgt gctgctcacg
      901 ccagcacaaa ccgaacagct gctgccagca ctgctgagcg acatcgacga gcagggtaaa
      961 ggtcgcgtca accgcgacta tgtcggccgt gatgccacga aactggctga agccatcggg
     1021 cttgaggtca acgaatatac ccgcttactg ctggctgaaa ccgacgccag ccacccgttt
     1081 gccgtcaccg agctgatgat gccggtacta cccgtggtgc gggtgaaaag cgtggatgac
     1141 gctattgcgc tggcactcaa actggaaaat ggctgccgcc acaccgccgc gatgcactcc
     1201 accaatatca ggaacttaaa tcgcatggct aacgccatca acaccagtat tttcgtcaaa
     1261 aatggccctt gtatcgccgg gttggggctg ggtggcgaag gttggacatc gatgaccatt
     1321 tcgacgccga ccggtgaagg tgtcacctct gctcgcacct ttgtgcgcct gcgacgctgt
     1381 gtgctggtgg atatgttccg tattgcctaa ccgttcattt atcacaaaag gattgttcga
     1441 tgtctattcc agaaactcaa aaagccatta tcttctacga atccaacggc aagttggagc
     1501 ataaggatat cccagttcca aagccaaagc ccaacgaatt gttaatcaac gtcaagtact
     1561 ctggtgtctg ccacaccgat ttgcacgctt ggcatggtga ctggccattg ccaactaagt
     1621 taccattagt tggtggtcac gaaggtgccg gtgtcgttgt cggcatgggt gaaaacgtta
     1681 agggctggaa gatcggtgac tacgccggta tcaaatggtt gaacggttct tgtatggcct
     1741 gtgaatactg tgaattgggt aacgaatcca actgtcctca cgctgacttg tctggttaca
     1801 cccacgacgg ttctttccaa gaatacgcta ccgctgacgc tgttcaagcc gctcacattc
     1861 ctcaaggtac tgacttggct gaagtcgcgc caatcttgtg tgctggtatc accgtataca
     1921 aggctttgaa gtctgccaac ttgagagcag gccactgggc ggccatttct ggtgctgctg
     1981 gtggtctagg ttctttggct gttcaatatg ctaaggcgat gggttacaga gtcttaggta
     2041 ttgatggtgg tccaggaaag gaagaattgt ttacctcgct cggtggtgaa gtattcatcg
     2101 acttcaccaa agagaaggac attgttagcg cagtcgttaa ggctaccaac ggcggtgccc
     2161 acggtatcat caatgtttcc gtttccgaag ccgctatcga agcttctacc agatactgta
     2221 gggcgaacgg tactgttgtc ttggttggtt tgccagccgg tgcaaagtgc tcctctgatg
     2281 tcttcaacca cgttgtcaag tctatctcca ttgtcggctc ttacgtgggg aacagagctg
     2341 ataccagaga agccttagat ttctttgcca gaggtctagt caagtctcca ataaaggtag
     2401 ttggcttatc cagtttacca gaaatttacg aaaagatgga gaagggccaa attgctggta
     2461 gatacgttgt tgacacttct aaatgatggg ctgaagggtt taagggctgc taacaaagcc
     2521 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     2581 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc
     2641 gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg
     2701 tgcctactaa gggcgaatgt acaggtttct acaaactctt ttgtttattt ttctaaatac
     2761 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2821 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2881 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2941 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3001 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3061 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3121 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3181 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3241 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3301 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3361 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3421 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3481 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3541 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3601 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3661 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3721 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3781 ataatctcat gaccaaaatc ccttaacgtt cacacaacat agccacggga gttttcgttc
     3841 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3901 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3961 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4021 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4081 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4141 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4201 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4261 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4321 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4381 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4441 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4501 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4561 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4621 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     4681 catctgtgcg gtatttcaca ccgcatatgc tggatcctaa catcatcatc accatcacta
     4741 aaaggaggcg acggcttcaa atttagcgat ctcttcgtgg tctacgttat ggttttccgg
     4801 cgatttttcg gcattcattg gcacttctac tccgtaattg gcaagacaaa cgagtatatc
     4861 aggcattgga tgtgaataaa gcgtataggt ttacctcaaa ctgcgcggct gtgttataat
     4921 ttgcgacctt tgaatccggg atactagaaa taat
//
