LOCUS       Exported                2178 bp ds-DNA     circular SYN 05-DEC-2018
DEFINITION  MoClo golden gate assembly AB part for pRpa-AM (Adam Meyer's 
            improved pRPA promoter from https://doi.org/10.1101/285866, selected
            for dynamic range and low cross-sensitivity)..
ACCESSION   .
VERSION     .
KEYWORDS    P66m
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2178)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 2178)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 5, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     overhang        1..4
                     /created_by="User"
                     /modified_by="murraylab"
                     /label="A Overhang"
     misc_feature    5..74
                     /Original_Bases="TTGACGGCTAGCTCAGTCCTAGGTACAGTGCTAGC"
                     /label="TTGACGGCTAGCTCAGTCCTAGGTACAGTGCTAGC"
                     /note="Geneious type: Editing History Replacement"
     promoter        5..74
                     /modified_by="murraylab"
                     /label="MURDOC-pRpa76-AM"
     misc_feature    complement(5..41)
                     /created_by="murraylab"
                     /Sequence="AATATTGTCCGATCGGACAGGTCAGCCAACGATCGTT"
                     /modified_by="murraylab"
                     /label="pRpa(AM) 1R"
                     /note="Geneious type: primer_bind_reverse"
     overhang        37..41
                     /created_by="murraylab"
                     /label="overlap"
     misc_feature    complement(42..78)
                     /created_by="murraylab"
                     /Sequence="AGTATTTATTCGAAAGTAACAAACCATTTTCTTGCGT"
                     /modified_by="murraylab"
                     /label="pRpa(AM) 2R"
                     /note="Geneious type: primer_bind_reverse"
     overhang        75..78
                     /created_by="User"
                     /modified_by="murraylab"
                     /label="B Overhang"
     misc_feature    79..2178
                     /ApEinfo_revcolor="#6ecff6"
                     /ApEinfo_fwdcolor="#6ecff6"
                     /label="DVLA"
     misc_feature    complement(80..85)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     misc_feature    complement(119..136)
                     /Mismatches=0
                     /created_by="primer3"
                     /annotation_group="24625307020399a3794ed7-0f69-45c6-874a-d
                     ad0fe2ad238"
                     /%GC="66.7"
                     /Tm="62.8"
                     /Hairpin_Tm="70.2"
                     /Self_Dimer_Tm="16.2"
                     /Pair_Dimer_Tm="10.4"
                     /Sequence="CGAGCGCAGCGAGTCAGT"
                     /Product_Size=166
                     /label="Rev Seq Primer MoClo Stage 1"
                     /note="Geneious type: primer_bind_reverse"
     misc_feature    2172..2177
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     misc_feature    2172..2177
                     /Recognition_pattern="GGTCTC(1/5)"
                     /annotation_group="GGTCTC"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     source          1..2178
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    86..106
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(112..129)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(283..871)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(363..382)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1042..1902)
                     /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     1665..1684
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1903..2007)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2075..2093
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      2097..2140
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2143..2164
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggagaacgat cgttggctga cctgtccgat cggacaatat tacgcaagaa aatggtttgt
       61 tactttcgaa taaatactag agacctacta gtagcggccg ctgcaggctt cctcgctcac
      121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
      181 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
      241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccaca ggctccgccc
      301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      721 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
      781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
      841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
      901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
      961 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1021 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1081 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1141 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgcgacccac gctcaccggc
     1201 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1261 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1321 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1381 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1441 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1501 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1561 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1621 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1681 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1741 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1801 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1861 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1921 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1981 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     2041 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggcagaattt
     2101 cagataaaaa aaatccttag ctttcgctaa ggatgatttc tggaattcgc ggccgcttct
     2161 agagactagt gggtctca
//
