LOCUS       Exported                2192 bp ds-DNA     circular SYN 05-DEC-2018
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    U9m
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2192)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 2192)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 5, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     misc_feature    1..2192
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     rbs             complement(5..88)
                     /created_by="murraylab"
                     /label="BCD22"
     misc_feature    93..2192
                     /ApEinfo_revcolor="#6ecff6"
                     /ApEinfo_fwdcolor="#6ecff6"
                     /label="DVLA"
     misc_feature    complement(94..99)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     primer_bind     complement(176..196)
                     /Sequence="TTACCGCCTTTGAGTGAGCTG"
                     /Hairpin_Tm="None"
                     /Tm="60.6"
                     /Self_Dimer_Tm="None"
                     /%GC="52.4"
                     /created_by="primer3"
                     /label="SC190/Part.Seq.REV"
     primer_bind     2088..2108
                     /Sequence="AAATAGGCGTATCACGAGGCA"
                     /Hairpin_Tm="46.5"
                     /Tm="59.6"
                     /Self_Dimer_Tm="None"
                     /%GC="47.6"
                     /created_by="primer3"
                     /label="SC189/Part.Seq.FOR"
     misc_feature    2186..2191
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     misc_feature    2186..2191
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     source          1..2192
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    100..120
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(126..143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(297..885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(377..396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1056..1916)
                     /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     1679..1698
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1917..2021)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2089..2107
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      2111..2154
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2157..2178
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tactgggccc aagttcactt aaaaaggaga tcaacaatga aagcaatttt cgtactgaaa
       61 catcttaatc atgcctagga agttttctaa tgagagacct actagtagcg gccgctgcag
      121 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      181 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      241 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      301 cacaggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      361 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      421 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      481 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      541 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      601 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      661 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      721 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      781 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      841 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      901 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      961 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1021 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1081 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1141 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgcgac
     1201 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1261 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1321 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1381 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1441 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1501 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1561 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1621 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1681 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1741 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1801 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1861 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1921 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1981 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2041 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2101 acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat
     2161 tcgcggccgc ttctagagac tagtgggtct ca
//
