LOCUS       sequence-238071-        2267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo golden gate assembly BC part for RiboJ-BCD2 (low expression 
            bi-cistronic RBS, engineered for context-independence, prefixed by 
            self-cleaving RiboJ; see https://doi.org/10.1038/nmeth.2404)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2267)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 2267)
  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: U24m
FEATURES             Location/Qualifiers
     source          1..2267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..28
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(32..75)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    78..99
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     misc_RNA        120..169
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             263..271
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     misc_feature    288..308
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(314..331)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(485..1073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(565..584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1244..2035,2036..2104))
                     /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     1867..1886
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2105..2209)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acctataaaa ataggcgtat cacgaggcag aatttcagat aaaaaaaatc cttagctttc
       61 gctaaggatg atttctggaa ttcgcggccg cttctagaga ctagtgggtc tcatactagc
      121 tgtcaccgga tgtgctttcc ggtctgatga gtccgtgagg acgaaacagc ctctacaaat
      181 aattttgttt aagggcccaa gttcacttaa aaaggagatc aacaatgaaa gcaattttcg
      241 tactgaaaca tcttaatcat gctaaggagg ttttctaatg agagacctac tagtagcggc
      301 cgctgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
      361 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
      421 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      481 cgtttttcca caggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      541 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      601 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      661 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      721 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      781 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      841 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      901 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      961 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1021 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1081 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1141 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1201 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1261 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     1321 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     1381 cgcgcgaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     1441 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     1501 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     1561 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     1621 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     1681 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     1741 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     1801 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     1861 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     1921 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     1981 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     2041 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     2101 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     2161 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     2221 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacatta
//
