LOCUS       Exported                2171 bp ds-DNA     circular SYN 04-DEC-2018
DEFINITION  MoClo golden gate assembly DE part for Eck120033736 (164.6x-strength
            natural terminator; see DOI: 10.1038/nmeth.2515).
ACCESSION   .
VERSION     .
KEYWORDS    T2m
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2171)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 2171)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 4, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     misc_feature    5..67
                     /Original_Bases="ATGTCTTCTGCACCATATGCGGTGTGAAATACCGCACAGAT
                     GCGTAAGGAGAAAATACCGCATCAGGCGCCATTCGCCATTCAGGCTGCGCAACTGTTG
                     GGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGGGGGATGT
                     GCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAA
                     CGACGGCCAGTGAATTCGAGCTCGGTACCCGGGGATCCTCTAGAGTCGACCTGCAGGC
                     ATGCAAGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCT
                     CACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAA
                     TGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAA
                     ACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAAGACGT"
                     /label="ATGTCTTCTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGA
                     GAAAATACCGCATCAGGCGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCG
                     ATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGGGGGATGTGCTGCAAGG
                     CGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGGCCA
                     GTGAATTCGAGCTCGGTACCCGGGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCT
                     TGGCGTAATCATGGTCATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCC
                     ACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGC
                     TAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGT
                     GCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAAGACGT"
                     /note="Geneious type: Editing History Replacement"
     terminator      7..59
                     /created_by="User"
                     /modified_by="User"
                     /label="ECK120033736"
     misc_feature    72..2171
                     /ApEinfo_revcolor="#6ecff6"
                     /ApEinfo_fwdcolor="#6ecff6"
                     /label="DVLA"
     misc_feature    complement(73..78)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     misc_feature    80..2157
                     /ApEinfo_revcolor="#6ecff6"
                     /ApEinfo_fwdcolor="#6ecff6"
                     /Transferred_From="DVA"
                     /Transferred_Similarity="100.00%"
                     /label="DVLA"
     primer_bind     complement(155..175)
                     /Sequence="TTACCGCCTTTGAGTGAGCTG"
                     /Hairpin_Tm="None"
                     /Tm="60.6"
                     /Self_Dimer_Tm="None"
                     /%GC="52.4"
                     /created_by="primer3"
                     /Transferred_From="DVA"
                     /Transferred_Similarity="100.00%"
                     /label="SC190/Part.Seq.REV"
     primer_bind     2067..2087
                     /Sequence="AAATAGGCGTATCACGAGGCA"
                     /Hairpin_Tm="46.5"
                     /Tm="59.6"
                     /Self_Dimer_Tm="None"
                     /%GC="47.6"
                     /created_by="primer3"
                     /Transferred_From="DVA"
                     /Transferred_Similarity="100.00%"
                     /label="SC189/Part.Seq.FOR"
     misc_feature    2165..2170
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     misc_feature    2165..2170
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     source          1..2171
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    79..99
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(105..122)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(276..864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(356..375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1035..1895)
                     /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     1658..1677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1896..2000)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2068..2086
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      2090..2133
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2136..2157
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 aggtctaacg catgagaaag cccccggaag atcaccttcc gggggctttt ttattgcgcc
       61 cttgagagct tagagaccta ctagtagcgg ccgctgcagg cttcctcgct cactgactcg
      121 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
      181 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
      241 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc acaggctccg cccccctgac
      301 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      361 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      421 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
      481 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
      541 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
      601 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      661 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
      721 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      781 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      841 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      901 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
      961 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1021 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     1081 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     1141 ttaccatctg gccccagtgc tgcaatgata ccgcgcgacc cacgctcacc ggctccagat
     1201 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     1261 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     1321 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     1381 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     1441 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     1501 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     1561 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     1621 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     1681 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     1741 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     1801 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     1861 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     1921 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     1981 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     2041 attattatca tgacattaac ctataaaaat aggcgtatca cgaggcagaa tttcagataa
     2101 aaaaaatcct tagctttcgc taaggatgat ttctggaatt cgcggccgct tctagagact
     2161 agtgggtctc a
//
