LOCUS       sequence-181872-        3737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of zmADH using the p70b (weak version of the p70a 
            promoter) promoter and UTR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3737)
  AUTHORS   Schlesinger O, Chemla Y, Heltberg M, Ozer E, Marshall R, Noireaux V,
            Jensen MH, Alfonta L
  TITLE     Tuning of Recombinant Protein Expression in Escherichia coli by 
            Manipulating Transcription, Translation Initiation Rates, and 
            Incorporation of Noncanonical Amino Acids.
  JOURNAL   ACS Synth Biol. 2017 Mar 9. doi: 10.1021/acssynbio.7b00019.
  PUBMED    28230975
REFERENCE   2  (bases 1 to 3737)
  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: "ACS Synth 
            Biol. 2017 Mar 9. doi: 10.1021/acssynbio.7b00019."
COMMENT     Sequence Label: pBEST-p70b-UTR1-NTerm 6xHis-zmADH-T500
FEATURES             Location/Qualifiers
     source          1..3737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             303..325
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             344..361
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             371..388
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1720..1737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1891..2480)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1971..1990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2651..3442,3443..3511))
                     /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     3274..3293
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3512..3616)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3684..3702
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ccagccagaa aacgaccttt ctgtggtgaa accggatgct gcaattcaga gcgccagcaa
       61 gtgggggaca gcagaagacc tgaccgccgc agagtggatg tttgacatgg tgaagactat
      121 cgcaccatca gccagaaaac cgaattttgc tgggtgggct aacgatatcc gcctgatgcg
      181 tgaacgtgac ggacgtaacc accgcgacat gtgtgtgctg ttccgctggg catgctgagc
      241 taacaccgtg cgtgtagaca attttacctc tggcggtgat aatggttgca gctagcaata
      301 attttgttta actttaagaa ggagatatac catgggcagc agccatcatc atcatcatca
      361 cagcagcggc ctggtgccgc gcggcagcca tatggcttct tcaacttttt atattccttt
      421 cgtcaacgaa atgggcgaag gttcgcttga aaaagcaatc aaggatctta acggcagcgg
      481 ctttaaaaat gccctgatcg tttctgatgc tttcatgaac aaatccggtg ttgtgaagca
      541 ggttgctgac ctgttgaaaa cacagggtat taattctgct gtttatgatg gcgttatgcc
      601 gaacccgact gttaccgcag ttctggaagg ccttaagatc ctgaaggata acaattcaga
      661 cttcgtcatc tccctcggtg gtggttctcc ccatgactgc gccaaagcca tcgctctggt
      721 cgcaaccaat ggtggtgaag tcaaagacta cgaaggtatc gacaaatcta agaaacctgc
      781 cctgcctttg atgtcaatca acacgacggc tggtacggct tctgaaatga cgcgtttctg
      841 catcatcact gatgaagtcc gtcacgttaa gatggccatt gttgaccgtc acgttacccc
      901 gatggtttcc gtcaacgatc ctctgttgat ggttggtatg ccaaaaggcc tgaccgccgc
      961 caccggtatg gatgctctga cccacgcatt tgaagcttat tcttcaacgg cagctactcc
     1021 gatcaccgat gcttgcgctt tgaaagcagc ttccatgatc gctaagaatc tgaagaccgc
     1081 ttgcgacaac ggtaaggata tgccagctcg tgaagctatg gcttatgccc aattcctcgc
     1141 tggtatggcc ttcaacaacg cttcgcttgg ttatgtccat gctatggctc accagttggg
     1201 cggttactac aacctgccgc atggtgtctg caacgctgtt ctgcttccgc atgttctggc
     1261 ttataacgcc tctgtcgttg ctggtcgtct gaaagacgtt ggtgttgcta tgggtctcga
     1321 tatcgccaat ctcggcgata aagaaggcgc agaagccacc attcaggctg ttcgcgatct
     1381 ggctgcttcc attggtattc cagcaaatct gaccgagctg ggtgctaaga aagaagatgt
     1441 gccgcttctt gctgaccacg ctctgaaaga tgcttgtgct ctgaccaacc cgcgtcaggg
     1501 tgatcagaaa gaagttgaag aactcttcct gagcgctttc taactcgagc aaagcccgcc
     1561 gaaaggcggg cttttctgtg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     1621 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     1681 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     1741 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1801 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1861 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1921 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1981 ggcgtttccc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2041 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2101 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2161 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2221 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2281 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2341 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2401 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2461 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2521 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2581 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2641 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2701 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2761 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2821 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2881 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2941 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3001 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3061 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3121 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3181 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3241 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt
     3301 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3361 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3421 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3481 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3541 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3601 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     3661 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat
     3721 tctggcgaat cctctga
//
