LOCUS       sequence-111948-        5364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NADH Dependent 2,3-BDO Production Construct, Low Phosphate Dependent
            E. coli Expression .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5364)
  TITLE     SUPERSEDE
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5364)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSMART-2,3-BDO1
FEATURES             Location/Qualifiers
     source          1..5364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      3641..3672
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3673..3775
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3776..4591
                     /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(3849..3868)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      4615..4642
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(4654..5241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4734..4753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5263..5292)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 gtgcgtaatt gtgctgatct cttatatagc tgctctcatt atctctctac cctgaagtga
       61 ctctctcacc tgtaaaaata atatctcaca ggcttaatag tttcttaata caaagcctgt
      121 aaaacgtcag gataacttct tggaaaaagg agatatacca tgatgcacag cagcgcatgt
      181 gattgtgaag cgagtttgtg cgagacactc cgtggttttt ccgcaaaaca cccggattcc
      241 gtaatctacc agacatcact gatgtccgcc cttctgtcag gcgtatatga aggggacacg
      301 actattgcgg atcttctggc ccacggcgat tttggcctgg gtacgttcaa tgaactcgac
      361 ggcgaaatga tcgcgttttc ttcgcaagtt tatcagctcc gtgcggatgg gagcgcccgc
      421 gccgcgaagc cagaacaaaa aacaccgttt gcagtaatga catggttcca accgcagtat
      481 cgtaaaactt tcgatgcccc ggtgagtcgt cagcagatcc acgatgtaat cgatcaacag
      541 attccttcag acaacctgtt ttgcgcgctg cgtattgacg ggaatttccg tcatgctcac
      601 acacgtaccg ttccgcgcca gaccccaccc tatcgcgcga tgaccgatgt gctggatgat
      661 caaccggtct ttcgttttaa ccagcgcgaa ggagttctgg tgggttttcg taccccgcaa
      721 catatgcagg gtattaacgt ggcgggctac catgagcatt tcattacaga tgatcgccaa
      781 ggcggtggtc acctgttgga ttaccagctg gaatctggcg tcctgacttt cggcgagatt
      841 cacaaactga tgattgacct gccggcggat tctgcattcc tgcaggcaaa tttgcacccc
      901 agcaaccttg atgccgccat ccgctccgtc gagaactaat aggctcttca cttctggaaa
      961 aaggagatat accatgaatt ccgaaaaaca atcgcgtcag tgggcacatg gtgctgatat
     1021 ggttgtgggc cagctggagg cgcagggggt taaacaggtc tttggtattc cgggtgctaa
     1081 gatcgacaaa gtgtttgatt ctttactgga tagctcaatc gagattatcc cggtgcgtca
     1141 tgaagcaaac gcagcgttca tggccgcggc agttggtcgc cttacgggta aagctggcgt
     1201 agccctggtc acaagcggcc ccgggtgctc gaatctcatt accggcattg caaccgcaaa
     1261 ttctgaggga gatcctgtag tggcactggg gggcgcggta aaacgtgctg ataaagcgaa
     1321 attagttcac cagagtatgg acaccgtcgc gatgttctct ccagtaacca aatatgcggt
     1381 tgaagtttct tccccagatg caattgcaga ggtagtatca aacgcttttc gtgccgcgga
     1441 acatggccgc ccaggtgggg cgttcgtttc gctgccgcag gatattgtag accaaccggc
     1501 gacaggcgcc atcctgcctg catctggccc ggcactgatg ggcccagcgc cagagtcggc
     1561 gattaacgat gtggcaaaac ttatcgacaa cgccaaaaac cctgtgattc tgttgggctt
     1621 aatggcatca cagccggcta attcggctgc attgcgtaag ctgctggaga agagtcgcat
     1681 cccggtgact tccacctacc aagccgccgg agctgtgaac caagaacatt tcacccgctt
     1741 cgccggtcgt gttggccttt tcaataacca agcgggagac cgtctgctgc atttggccga
     1801 tctcattatc tgtattggat actctccagt cgagtatgaa ccgagcatgt ggaactcggg
     1861 tgacgcaacc ctcgttcata ttgacgtgct gccagcttat gaagaacgca actatgtacc
     1921 cgatatcgag ttggtaggcg acattgcggc gacactgaac ctgctcgctt cccgcattga
     1981 tcataaactg gagctctcgc agcgtgcctc cgagatctta gtcgatcgcc aacaccagcg
     2041 cgatctgctg gatcgccgtg gcgcaagctt aaatcaattt gcgctgcatc cattacgtat
     2101 cgtccgtgcc atgcaggaca tcgtaaacaa tgacgtaacg ctgaccgtgg acatgggctc
     2161 atttcatatt tggatcgcac gctatctcta ttcatttcgc gcacgtcagg tcatgattag
     2221 taatgggcaa caaactatgg gcgtggctct gccttgggct atcggtgcgt ggctggtgaa
     2281 ccccggccgc aaagtggtga gcgttagcgg tgacggagga tttctgcaga gtagcatgga
     2341 gttagaaacc gctgtccgcc tgaacgctaa tgtgttacac atcatttggg tggataatgg
     2401 ttataatatg gttgcaatcc aggaggagaa aaagtatcag cgtttaagcg gtgtggcgtt
     2461 tggaccggta gatttcaaag cctacgccga tgcattcggc gcccgtggct tcgcggtcga
     2521 aagcgcggat gccttagaga gcaccttacg tgcggcaatg gatgtgaatg gtccggccgt
     2581 cgtggcgatt ccggtggatt attcggataa tccgctgctg atgggacaac tgcacctttc
     2641 gcagatcctg tagtaagctc ttctggaaaa aggagatata ccatgcagaa ggtggcgctc
     2701 gttaccggat ctggccaagg cattggcaaa gcgattgcgc ttcgtctggt caaagacgga
     2761 ttcgccgttg caattgctga ttacaacgac gaaacggcgc gtgctgtcgc cgatgaaatc
     2821 atccgtaatg gtggcaacgc tgtcgcagtg aaagtggacg tctctgatcg cgaccaagta
     2881 tttgcagcgg tcgagaaagc acgtaccgct ctgggcggtt tcaacgttat cgtgaacaac
     2941 gcgggcattg cgccgtcgac gcctatcgaa agcatcaccc cggagattgt agataaggtg
     3001 tacaacatca acgtaaaagg agtaatctgg ggtatgcaag ctgccatcga tgcgttccgc
     3061 aaagaggggc acggcggtaa aatcattaac gcgtgttcgc aggctggtca tactggtaac
     3121 ccggaactgg cggtttatag cagcagcaaa ttcgccgtgc gtggcctgac ccagaccgct
     3181 gcacgcgatc tggcgccgct ggggatcacc gtcaatgcat attgtccggg tatcgtaaaa
     3241 accccgatgt gggcggaaat tgatcgccag gtatcagagg ccgctggcaa accgctgggc
     3301 tatggcacgg aaacgtttgc caagcgcatc acgttaggcc gtctgtcgga accggaggat
     3361 gttgcagcat gcgtctctta cctggcgggc ccggattctg attatatgac gggtcagtcc
     3421 ctgctgattg atggtggcat ggtctttaac tagtaagatc gtcccggctt atcggtcagt
     3481 ttcacctgat ttacgtaaaa acccgcttcg gcgggttttt gcttttggag gggcagaaag
     3541 atgaatgact gtccacgacg ctatacccaa aagaaagacg aattctctag atatcgctca
     3601 atactgacca tttaaatcat acctgacctc catagcagaa agtcaaaagc ctccgaccgg
     3661 aggcttttga cttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
     3721 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgag
     3781 ccatattcaa cgggaaacgt cttgctcgag gccgcgatta aattccaaca tggatgctga
     3841 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     3901 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     3961 caatgatgtt acagatgaga tggtcaggct aaactggctg acggaattta tgcctcttcc
     4021 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     4081 agggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     4141 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     4201 cggcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttgg
     4261 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     4321 gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     4381 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     4441 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     4501 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     4561 gtttcacttg atgctcgatg agtttttcta atgagggccc aaatgtaatc acctggctca
     4621 ccttcgggtg ggcctttctg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4681 gcatcacaaa aatcgatgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4741 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4801 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4861 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4921 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4981 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5041 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5101 atttggtatc tgcgctctgc tgaagccagt tacctcggaa aaagagttgg tagctcttga
     5161 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5221 cgcagaaaaa aaggatctca agaagatcct ttgattttct accgaagaaa ggcccacccg
     5281 tgaaggtgag ccagtgagtt gattgcagtc cagttacgct ggagtctgag gctcgtcctg
     5341 aatgatatca agcttgaatt cgtt
//
