LOCUS       sequence-147244-        2274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insulated MoClo promoter for E. coli. 36nt DNA spacer empirically 
            selected. Modified from BBa_I13453. Has near identical expression as
            36INS1-pBAD_AB [G:36INS2-pBAD:B].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2274)
  AUTHORS   Carr SB, Beal J, Densmore DM
  TITLE     Reducing DNA context dependence in bacterial promoters.
  JOURNAL   PLoS One. 2017 Apr 19;12(4):e0176013. doi: 
            10.1371/journal.pone.0176013. eCollection 2017.
  PUBMED    28422998
REFERENCE   2  (bases 1 to 2274)
  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: "PLoS One.";
            date: "2017-04-19"; volume: "12(4):e0176013. doi"; pages: " 
            10.1371/journal.pone.0176013. eCollection 2017"
COMMENT     Sequence Label: 36INS2-pBAD_GB
FEATURES             Location/Qualifiers
     source          1..2274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     74..93
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     misc_feature    182..202
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(208..225)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(379..967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(459..478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1138..1929,1930..1998))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1761..1780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1999..2103)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2171..2189
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2193..2236)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2239..2260
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tgcctagcac gcttttctgc gtatggttgg tcagcttcta acattgatta tttgcacggc
       61 gtcacacttt gctatgccat agcattttta tccataagat tagcggatcc tacctgacgc
      121 tttttatcgc aactctctac tgtttctcca taccgttttt ttgggctagc tactagagac
      181 ctactagtag cggccgctgc aggcttcctc gctcactgac tcgctgcgct cggtcgttcg
      241 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      301 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      361 ggccgcgttg ctggcgtttt tccacaggct ccgcccccct gacgagcatc acaaaaatcg
      421 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      481 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      541 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      601 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      661 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      721 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      781 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      841 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      901 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      961 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1021 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1081 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1141 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1201 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1261 tgctgcaatg ataccgcgcg acccacgctc accggctcca gatttatcag caataaacca
     1321 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1381 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1441 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1501 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1561 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1621 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1681 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1741 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1801 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1861 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1921 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1981 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2041 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2101 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     2161 aacctataaa aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt
     2221 cgctaaggat gatttctgga attcgcggcc gcttctagag actagtgggt ctca
//
