LOCUS       sequence-304537-        4952 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 4952)
  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 4952)
  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..4952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      130..718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     619..638
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     872..889
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             969..986
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1203..1225
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3751..3769)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3765..3812
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3942..4033
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4034..4102,4103..4894)
                     /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(4252..4271)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtc
       61 acacaacata gccacggagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      121 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      181 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      241 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
      301 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      361 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      421 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      481 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      541 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      601 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      661 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      721 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      781 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      841 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      901 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatgctg
      961 gatcctaaca tcatcatcac catcactaaa aggaggcgac ggcttcaaat ttagcgatct
     1021 cttcgtggtc tacgttatgg ttttccggcg atttttcggc attcattggc acttctactc
     1081 cgtaattggc aagacaaacg agtatatcag gcattggatg tgaataaagc gtataggttt
     1141 acctcaaact gcgcggctgt gttataattt gcgacctttg aatccgggat actagaaata
     1201 attttgttta actttaagaa ggagatatac atatggaaca ctccgtgatc gaaccgactg
     1261 taccgatgcc catgccagcg atgtttgacg caccgagcgg cattttcgac tcgcttgacg
     1321 acgccgttca ggccgccgtg tacgcgcaac aacagttgaa cagcgtcgag ctgcgtcaac
     1381 aggtgattaa agccattcgg gtcgctggcg agcgttatgc ccaggtgctg gcggaaatgg
     1441 ccgtggccga aaccggcatg ggccgcgtgg tcgataagta catcaaaaac gtgtcacagg
     1501 cgcgccatac accagggatc gaatgcctgt cggcggaagt actgaccggt gataatgggc
     1561 tgacgctgat tgaaaatgcc ccctggggtg tggttgcgtc ggtcaccccg tcaaccaacc
     1621 cggccgcaac cgttatcaat aacgccatca gtatgatagc agccggtaac agcgtggtgt
     1681 tcgcccctca tccgtcagcg aaaaacgtat cgctgcgtac catcagcctg ctcaataagg
     1741 ccatcgtcgc caccggcggc ccggagaacc tgctggtcag cgtttccgac cccaacatcg
     1801 aaaccgcgca acgcctgttt cgctaccccg gtatcgggct gctggttgtc accggcggcg
     1861 aagcggtggt ggaagcggcg cgcagacata ccgacaaacg gctgatcgcc gccggagcgg
     1921 gtaacccgcc agtggtggtg gatgaaacgg cggatattcc gaaagcagcc cgcgcgattg
     1981 ttaaaggcgc ctcattcgac aacaacatta tctgtgccga tgagaaggtg ctgatcgtgg
     2041 tggacagtgt ggccgatgcc ctactggcag aaatgcaacg caatcatgcc gtgctgctca
     2101 cgccagcaca aaccgaacag ctgctgccag cactgctgag cgacatcgac gagcagggta
     2161 aaggtcgcgt caaccgcgac tatgtcggcc gtgatgccac gaaactggct gaagccatcg
     2221 ggcttgaggt caacgaatat acccgcttac tgctggctga aaccgacgcc agccacccgt
     2281 ttgccgtcac cgagctgatg atgccggtac tacccgtggt gcgggtgaaa agcgtggatg
     2341 acgctattgc gctggcactc aaactggaaa atggctgccg ccacaccgcc gcgatgcact
     2401 ccaccaatat caggaactta aatcgcatgg ctaacgccat caacaccagt attttcgtca
     2461 aaaatggccc ttgtatcgcc gggttggggc tgggtggcga aggttggaca tcgatgacca
     2521 tttcgacgcc gaccggtgaa ggtgtcacct ctgctcgcac ctttgtgcgc ctgcgacgct
     2581 gtgtgctggt ggatatgttc cgtattgcct aaccgttcat ttatcacaaa aggattgttc
     2641 gatgtctatt ccagaaactc aaaaagccat tatcttctac gaatccaacg gcaagttgga
     2701 gcataaggat atcccagttc caaagccaaa gcccaacgaa ttgttaatca acgtcaagta
     2761 ctctggtgtc tgccacaccg atttgcacgc ttggcatggt gactggccat tgccaactaa
     2821 gttaccatta gttggtggtc acgaaggtgc cggtgtcgtt gtcggcatgg gtgaaaacgt
     2881 taagggctgg aagatcggtg actacgccgg tatcaaatgg ttgaacggtt cttgtatggc
     2941 ctgtgaatac tgtgaattgg gtaacgaatc caactgtcct cacgctgact tgtctggtta
     3001 cacccacgac ggttctttcc aagaatacgc taccgctgac gctgttcaag ccgctcacat
     3061 tcctcaaggt actgacttgg ctgaagtcgc gccaatcttg tgtgctggta tcaccgtata
     3121 caaggctttg aagtctgcca acttgagagc aggccactgg gcggccattt ctggtgctgc
     3181 tggtggtcta ggttctttgg ctgttcaata tgctaaggcg atgggttaca gagtcttagg
     3241 tattgatggt ggtccaggaa aggaagaatt gtttacctcg ctcggtggtg aagtattcat
     3301 cgacttcacc aaagagaagg acattgttag cgcagtcgtt aaggctacca acggcggtgc
     3361 ccacggtatc atcaatgttt ccgtttccga agccgctatc gaagcttcta ccagatactg
     3421 tagggcgaac ggtactgttg tcttggttgg tttgccagcc ggtgcaaagt gctcctctga
     3481 tgtcttcaac cacgttgtca agtctatctc cattgtcggc tcttacgtgg ggaacagagc
     3541 tgataccaga gaagccttag atttctttgc cagaggtcta gtcaagtctc caataaaggt
     3601 agttggctta tccagtttac cagaaattta cgaaaagatg gagaagggcc aaattgctgg
     3661 tagatacgtt gttgacactt ctaaatgatg ggctgaaggg tttaagggct gctaacaaag
     3721 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     3781 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc
     3841 ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac
     3901 ggtgcctact aagggcgaat gtacaggttt ctacaaactc ttttgtttat ttttctaaat
     3961 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4021 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4081 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4141 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4201 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4261 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     4321 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4381 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4441 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4501 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4561 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4621 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4681 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4741 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4801 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4861 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4921 actttagatt gatttaaaac ttcattttta at
//
