LOCUS       sequence-339045-        4073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative control pBAD backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4073)
  AUTHORS   Ho JML, Miller CA, Smith KA, Mattia JR, Bennett MR
  TITLE     Improved pyrrolysine biosynthesis through phage assisted 
            non-continuous directed evolution of the complete pathway.
  JOURNAL   Nat Commun. 2021 Jun 24;12(1):3914. doi: 10.1038/s41467-021-24183-9.
  PUBMED    34168131
REFERENCE   2  (bases 1 to 4073)
  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: "Nat 
            Commun."; date: "2021-06-24"; volume: "12(1):3914. doi"; pages: " 
            10.1038/s41467-021-24183-9"
COMMENT     Sequence Label: pJH572
FEATURES             Location/Qualifiers
     source          1..4073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      39..66
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        85..176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(177..245,246..1037)
                     /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(395..414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1208..1796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1697..1716
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1961..1978
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1993..2133
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2059..2078)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2219..2241
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2397..3275)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3302..3586
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     3509..3528
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(3764..3781)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3934..4020
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
       61 cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc
      121 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      181 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      241 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      301 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
      361 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
      421 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      481 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      541 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      601 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      661 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      721 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
      781 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
      841 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
      901 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
      961 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1021 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     1081 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1141 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1201 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1261 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1321 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     1381 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1441 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1501 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1561 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1621 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1681 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1741 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1801 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct ggccttttgc
     1861 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1921 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     1981 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2041 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     2101 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     2161 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     2221 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagcagatc
     2281 aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct gtcaaatgga cgaagcaggg
     2341 attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt taccaattat
     2401 gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg
     2461 ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca
     2521 ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg
     2581 atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt
     2641 gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg
     2701 tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga
     2761 tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc
     2821 gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg
     2881 tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt
     2941 gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga
     3001 taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc
     3061 aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg
     3121 cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg
     3181 agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat
     3241 cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc gccattcaga
     3301 gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc
     3361 ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc
     3421 aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt
     3481 gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg
     3541 gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc gttttttggg
     3601 ctaacaggag gaattaacca tgattaagaa gatggcgact gaagatctgg gtattggaac
     3661 tgcttctatg ctgtatagca ttttgtaaag ctcgagatct gcagctggta ccatatggga
     3721 attcgaagct tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat
     3781 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
     3841 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
     3901 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     3961 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     4021 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggc
//
