LOCUS       sequence-236185-        2196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo golden gate assembly BC part for BCD22 (very low expression 
            bi-cistronic RBS, engineered for downstream context-independence; 
            see https://doi.org/10.1038/nmeth.2404)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2196)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 2196)
  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"
COMMENT     Sequence Label: U9m
FEATURES             Location/Qualifiers
     source          1..2196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..33
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(37..80)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    83..104
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     misc_feature    222..242
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(248..265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(419..1007)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(499..518)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1178..1969,1970..2038))
                     /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     1801..1820
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2039..2143)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattaaccta taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag
       61 ctttcgctaa ggatgatttc tggaattcgc ggccgcttct agagactagt gggtctcata
      121 ctgggcccaa gttcacttaa aaaggagatc aacaatgaaa gcaattttcg tactgaaaca
      181 tcttaatcat gcctaggaag ttttctaatg aatgagagac ctactagtag cggccgctgc
      241 aggcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
      301 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
      361 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      421 tccacaggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
      481 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
      541 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
      601 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      661 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      721 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
      781 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
      841 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
      901 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
      961 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1021 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1081 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     1141 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     1201 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     1261 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgcg
     1321 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     1381 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1441 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     1501 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1561 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1621 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1681 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     1741 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1801 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     1861 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     1921 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     1981 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     2041 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2101 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     2161 tgccacctga cgtctaagaa accattatta tcatga
//
